43 results on '"D. A. Khochenkov"'
Search Results
2. Immunohistochemical analysis of prame expression as a prognostic factor in uveal melanoma patients
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S. V. Saakyan, A. Yu. Tsygankov, A. G. Amiryan, M. R. Khlgatyan, D. A. Khochenkov, and V. A. Misyurin
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увеальная меланома ,иммуногистохимическое исследование ,экспрессия ,prame ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
The study objective is to analyze the prognostic significance of PRAME protein expression in patients with uveal melanoma using immunohistochemical assay.Materials and methods. A total of 30 patients with uveal melanoma were examined and treated. The average age of patients at the time of treatment was 51.3 ± 11.8 years. In all cases, enucleation was performed according to the indications. A routine clinical-morphological, molecular-genetic, and immunohistochemical assay study was performed (n = 29). The immunohistochemical study was performed using antibodies to PRAME (clone 6H8, dilution 1: 50). The median follow-up period was 86.3 ± 2.9 months.Results. Of the 29 samples studied, staining was determined in 16, which was 55.2 %. Weak intensity of 1+ staining (from 10 to 20 % of tumor cells) was detected in 7 uveal melanoma samples, medium intensity of 2+ (from 10 to 20 % of tumor cells) – also in 7, and strong intensity of 3+ (30 % of tumor cells) – in 2 tumor samples. When assessing the seven-year survival, the cumulative survival rate in the group without PRAME expression was 0.857, while in the group with PRAME expression it was significantly lower and amounted to 0.357 (p = 0.0001). The PRAME protein expression was significantly correlated with the epithelioid cell type of the tumor (p = 0.041) and with the total and partial monosomy of chromosome 3 (p = 0.013).Conclusion. This paper presents the world’s first study of the prognostic significance of PRAME protein expression by immunohistochemical analysis in patients with uveal melanoma. A significant association of PRAME-positive patients with an unfavorable vital prognosis was shown.
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- 2021
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3. EXPRESSION OF THE VASCULAR ENDOTHELIAL GROWTH FACTOR AND ITS RECEPTORS (VEGFR-1 AND VEGFR-2) IN PRIMARY TUMOR CELLS IN PATIENTS WITH RENAL CANCER
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D. A. Khochenkov, M. I. Volkova, I. V. Timofeev, A. S. Olshanskaya, Yu. A. Khochenkova, E. Sh. Solomko, S. A. Ashuba, and V. B. Matveev
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почечно-клеточная карцинома ,рак почки ,фактор роста эндотелия сосудов ,экспрессия рецепторов ,общая выживаемость ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Purpose: to study the expression of vascular endothelial growth factor (vegf-a) and its receptors (vegfr-1 and vegfr-2) in renal cell carcinoma (rcc) cells and assess the effect of the expression levels of these markers on the tumor characteristics and prognosis of patients with rcc.Material and methods. The study included 65 patients with rcc (pt1a-t4n0/+m0/+). All patients underwent radical surgery. Histological tumor tissue samples obtained during surgery were used for the study. Expression of vegfa, vegfr-1, -2 was studied by immunohistochemical staining using appropriate antibodies to receptors and growth factors.Results. Expression of vegf and vegfr-1 and vegfr-2 receptors was ound in the cytoplasm and on the membrane of primary tumor cells of patients with rcc. There was a significant direct correlation of overexpression of the markers with g 3-4 anaplasia (vegfr-1, -2) and signs of significant tumor extension, including high pt category (vegfr-1, -2), larger size of the primary tumor (vegfr-1 , -2), tumor invasion of paranephria (vegf, vegfr-1), tumor venous thrombosis (vegfr-1, -2), multiple metastases (vegf-2), metastases in the adrenal gland (vegf, vegfr-2) and liver (vegfr-1) (p0.05).Conclusion. Expression of vegfa, as well as vegfr-1 and vegfr-2 receptors, was found on the surface and in the cytoplasm of cells of the primary tumor of patients with rcc (pt1a-t4n0/+m0/+). There was a significant correlation between vegf/vegfr overexpression with a high grade (g3-4) tumor anaplasia and significant tumor extension. In univariate analysis, a significant adverse effect on specific survival of vegfr-2 overexpression was observed. In regression analysis, vegfr-2 overexpression tended to independently affect specific survival. These results show the importance of vegf/vegfr expression as biomarkers in renal cell carcinoma.
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- 2021
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4. In vitro и in vivo photodynamic therapy of solid tumors with a combination of riboflavin and upconversion nanoparticles
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N. V. Sholina, R. A. Akasov, D. A. Khochenkov, A. N. Generalova, V. A. Semchishen, and E. V. Khaydukov
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riboflavin ,upconversion nanoparticles ,photodynamic therapy ,photosensitizer ,near infrared light ,Medicine - Abstract
Rationale: Riboflavin (vitamin B2) is one of the most promising agents for photodynamic therapy (PDT). However, its use is limited by the excitation in the ultraviolet (UV) and visible spectral ranges and, as a result, by a small penetration into biological tissue not exceeding a few millimeters. This problem could be solved by approaches ensuring excitation of riboflavin molecules within tumor tissues by infrared (IR) light. Upconversion nanoparticles (UCNPs) can be potentially considered as mediators able to effectively convert the exciting radiation of the near IR range, penetrating into biological tissue to a 3 cm depth, into the photoluminescence in the UV and visible spectral ranges.Aim: To evaluate the efficacy of UCNPs for IR-mediated riboflavin activation in the depth of tumor tissue during PDT. Materials and methods: The water-soluble riboflavin flavin mononucleotide (FMN, Pharmstandard-UfaVITA, Russia) was used as a photosensitizer in in vitro and in vivo experiments. The in vitro experiments were performed on human breast adenocarcinoma SK-BR-3, human glioblastoma U-87 MG, and rat glioma C6 cell lines. Lewis lung carcinoma (LLC) inoculated to hybrid BDF1 mice was used as a model to demonstrate the delivery of FMN to the tumor. UCNPs with a core/shell structure [NaYF4:Yb3+, Tm3+/NaYF4] were used for photoactivation of FMN in vivo. PDT based on FMN, UCNPs and laser radiation 975 nm (IR) was performed on mouse xenografts of human breast adenocarcinoma SKBR-3.Results: We were able to show that FMN could act as an effective in vitro photosensitizer for SK-BR-3, U-87 MG, and C6 cell lines. FMN IC50 values for glioma cells were ~30 μM, and for SK-BR-3 cell line ~50 μM (24 h incubation, irradiation 4.2 J/cm2). In the LLC model, the appropriate concentration of FMN (30 μM and above) can be achieved in the tumor as a result of systemic administration of FMN (at 2 and 24 hours after injection). The effect of PDT using near IR light for UCNP-mediated excitation of FMN was demonstrated in mouse xenografts SKBR-3, with the tumor growth inhibition of 90±5%.Conclusion: The study has demonstrated the possibility to use riboflavin (vitamin B2) as a photosensitizer for PDT. The photoexcitation of FMN via the anti-Stokes photoluminescence of UCNPs allows for implementation of the PDT technique with the near IR spectral range.
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- 2019
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5. New approaches in 3D modeling of in vitro growth of primary cultures of malignant gliomas
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Yu. A. Khochenkova, I. G. Dyrda, Yu. S. Machkova, E. Sh. Solomko, T. A. Sidorova, D. A. Khochenkov, and E. A. Avilova
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глиобластома ,3d-культура ,рецепторные тирозинкиназы ,темозоломид ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Background. The incidence of brain gliomas firmly occupies a leading position among all central nervous system tumors – 40–50 % of the cases detected, more than half of them are glioblastoma. Existing cell lines and cultivation methods do not reflect all the features of the three-dimensional (3D) organization of native glioblastoma. The use of temozolomide leads to the development of drug resistance and acute relapse, followed by a poor clinical outcome. The development of resistance is largely associated with the presence of tumor stem cells in the population and intratumoral heterogeneity. Obtaining 3D cultures from the primary material will allow us to save the stem cell pool and tumor-specific features.The study objective. Get a 3D model based on primary cell cultures, which allows you to save a heterogeneous population and the original phenotype of tumor cells.Materials and methods. We used U-87MG human glioma cells and GBM002 primary cell culture obtained from surgical material with a confirmed diagnosis of glioblastoma. Neurospheres were obtained from cell lines, the growth of which was monitored using the InCell Analyzer 6000 automatic cell analysis system. Flow cytometry was used to determine the CD133+ cell content. The expression of the receptor tyrosine kinases VEGFR1, VEGFR2 (endothelial growth factor type 1 and 2 receptors), FGFR2 (fibroblast growth factor receptor type 2) and the hypoxia marker HIF-1α (hypoxia inducible factor, 1α) in the neurospheres was evaluated using confocal microscopy.Results. GBM002 glioblastoma cells isolated from the surgical material formed neurospheres, while the number of CD133+ cells increased from 1–2 to 16–19 % compared with two-dimensional cultures. During long-term cultivation of cells with non-cytotoxic doses of temozolomide, it was found that such cells form smaller neurospheres compared to control cells. It was shown that the expression of receptor tyrosine kinases during cultivation of GBM002 glioblastoma cells in neurospheres differs from that in two-dimensional cultures. We found that in neurospheres, the expression of FGFR2 and VEGFR1, is significantly increased.Conclusion. 3D cultivation of primary cultures allows one to obtain a more heterogeneous population of tumor cells that reflects the spatial heterogeneity of cells, increase the pool of stem cells and recreate hypoxia conditions inside the brain micro-tumors.
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- 2019
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6. Photodynamic therapy of melanoma by blue-light photoactivation of flavin mononucleotide
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R. A. Akasov, N. V. Sholina, D. A. Khochenkov, A. V. Alova, P. V. Gorelkin, A. S. Erofeev, A. N. Generalova, and E. V. Khaydukov
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Medicine ,Science - Abstract
Abstract Melanoma is one of the most aggressive and lethal form of cancer. Photodynamic therapy (PDT) is a clinically approved technique for cancer treatment, including non-melanoma skin cancer. However, the most of conventional photosensitizers are of low efficacy against melanoma due to the possible dark toxicity at high drug concentrations, melanin pigmentation, and induction of anti-oxidant defense mechanisms. In the current research we propose non-toxic flavin mononucleotide (FMN), which is a water-soluble form of riboflavin (vitamin B2) as a promising agent for photodynamic therapy of melanoma. We demonstrated selective accumulation of FMN in melanoma cells in vivo and in vitro in comparison with keratinocytes and fibroblasts. Blue light irradiation with dose 5 J/cm2 of melanoma cells pre-incubated with FMN led to cell death through apoptosis. Thus, the IC50 values of human melanoma A375, Mel IL, and Mel Z cells were in a range of FMN concentration 10–30 µM that can be achieved in tumor tissue under systemic administration. The efficiency of reactive oxygen species (ROS) generation under FMN blue light irradiation was measured in single melanoma cells by a label-free technique using an electrochemical nanoprobe in a real-time control manner. Melanoma xenograft regression in mice was observed as a result of intravenous injection of FMN followed by blue-light irradiation of tumor site. The inhibition of tumor growth was 85–90% within 50 days after PDT treatment.
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- 2019
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7. THE ROLE OF EPITHELIAL-TO-MESENCHYMAL TRANSITION AND AUTOPHAGY IN ANTITUMORAL RESPONSE OF MELANOMA CELL LINES TO TARGET INHIBITION OF MEK AND mTOR KINASES
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O. O. Ryabaya, A. A. Prokofieva, D. A. Khochenkov, R. A. Akasov, S. V. Burov, E. A. Markvicheva, and E. V. Stepanova
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меланома ,аутофагия ,эпителиально-мезенхимальный переход ,3d-сфероиды ,экспериментальная терапия ,рапамицин ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Introduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kinases is a potential target for melanoma therapy by downregulatoin of the EMT.Objective: to study the effect of MEK and mTOR co-inhibition on cell viability, ability to form 3D-spheroids and migratory capacity of melanoma cell lines, and correlation of these changes with EMTand autophagy-related markers.Material and Methods. Melanoma cell lines Mel Z and Mel MTP were derived from patients, who were treated at the N.N. Blokhin National Medical Research Center of Oncology. The antiproliferative effect of binimetinib and/or rapamycin was studied by the MTT -test. 3D spheroids were formed using RGD peptides. Cell migration and invasion were assessed by a Boyden chamber migration assay. The expression levels of autophagy and EMT markers were investigated by immunocytochemistry or immunoblotting.Results. Rapamycin increased cytotoxicity of binimetinib in both 2D and 3D melanoma cell line cultures. At the same time, binimetinib and rapamycin reduced invasion, but not migration capacity of melanoma cells in vitro. The effectiveness of the combination was associated with a decrease in the EMT markers (N-cadherin and β-catenin) and autophagy markers (Beclin 1, p62/SQST M1 and LC3BII ) in melanoma cells.Conclusion. Inactivation of autophagy and EMT leads to overcoming the resistance to current anti-melanoma therapy and can be considered as a promising target for the treatment of melanoma.
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- 2019
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8. The role of autophagy inhibition in the enhanced cytotoxicity of temozolomide on melanoma cell lines
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O. O. Ryabaya, A. N. Inshakov, A. A. Malysheva, I. S. Abramov, N. V. Sholina, D. A. Khochenkov, and E. V. Stepanova
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меланома ,аутофагия ,апоптоз ,химиорезистентность ,темозоломид ,хлорокин ,ly-294.002 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Background. Despite advantages in treatment of metastatic melanoma it remains resistant to current therapy. Recent evidence indicates that tumor cells could overcome death through autophagy, a process that degrades cellular proteins and organelles to maintain cellular biosynthesis during nutrient deprivation or lack of energy. Objective: to investigate the involvement of autophagy inhibitors chloroquine (CQ) and LY-294.002 (LY) in temozolomide (TMZ) cytotoxicity in human melanoma cell lines.Materials and methods. The study was performed on patient-derived melanoma cell lines Mel Z, Mel IL and Mel MTP. The antiproliferative activity of combined TMZ and autophagy inhibitors treatment was determined by MTT assay and colony-forming assay. Cell cycle analysis, apoptosis activation and expression analysis of key autophagy markers under combined treatment was evaluated.Results. CQ and LY enhanced the cytotoxicity of TMZ and reduced colony formation in 3 melanoma cell lines, moreover both inhibitors increased cell population in G0 / G1 phase of cell cycle in Mel Z, Mel IL cell lines, but not in Mel MTP. CQ and LY synergistically activated apoptosis in all cell lines. The matrix RNA expression analysis of key autophagy genes showed autophagy involvement in enhanced cytotoxicity.Conclusions. Thus, autophagy inhibition on different stages of this process could overcome resistance to TMZ and be applicable as potent target in metastatic melanoma treatment.
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- 2017
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9. iNOS expression and biosynthesis of nitric oxide metabolites in the course of tumor growth of different histogenesis
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V. P. Deryagina, N. I. Ryzhova, L. V. Krivosheeva, I. S. Golubeva, L. A. Savluchinskaya, and D. A. Khochenkov
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биосинтез ,метаболиты оксида азота ,нитриты ,нитраты ,нитрозамины ,макрофаги ,экспрессия inos ,перевиваемые опухоли ,спонтанные опухоли ,химически индуцированные опухоли ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
The dynamics of the production of nitric oxide (NO) metabolites: nitrites, nitrates, volatile nitrosamines and iNOS expression was studied in mice with subcutaneous transplanted, spontaneous and chemical- induced tumors. Tumor growth was accompanied by increased production of nitrites + nitrates in tumors or their release with urine that not dependent on tumor histotype. The total concentration of nitrites and nitrates in tumors reached micromolar levels characteristic of nitrosative stress. The ability of peritoneal macrophages + monocytes to generates nitrites was suppressed at the stage of intensive growth of the Lewis lung carcinoma, which may indicate a decrease in the cytotoxic properties of immune cells. The possibility of formation in the Erlich carcinoma of volative N-nitrosodimethylamine and N-nitrosodiethylamine compounds with pronounced carcinogenic properties was demonstrated. A positive expression of iNOS was revealed in some areas of lung carcinoma at all investigated time points using the immunohistochemical method. The lungs metastases were not stain or weakly stained. This may indicate selection of the cells with a low activity of iNOS migrating in the lungs.
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- 2016
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10. LUMINESCENCE DIAGNOSTICS OF TUMORS WITH UPCONVERSION NANOPARTICLES
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V. V. Rocheva, N. V. Sholina, S. P. Derevyashkin, A. N. Generalova, A. V. Nechaev, D. A. Khochenkov, V. A. Semchishen, E. V. Khaydukov, E. V. Stepanova, and V. Ya. Panchenko
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upconverted nanoparticles ,optical luminescence imaging ,intraoperative assessment of tumor borders ,Medicine - Abstract
Background: To improve quality of surgery in oncology, it is necessary to completely remove the tumor, including its metastases, to minimize injury to normal tissues and to reduce duration of an intervention. Modern methods of detection based on radiological computerized tomography and magnetic resonance imaging can identify a tumor after its volume has become big enough, i.e. it contains more than 10 billion cells. Therefore, an improvement of sensitivity and resolution ability of diagnostic tools to identify early stages of malignant neoplasms seems of utmost importance. Aim: To demonstrate the potential of a new class of anti-Stokes luminescence nanoparticles for deep optical imaging with high contrast of malignant tumors. Materials and methods: Upconversion nanoparticles with narrow dispersion and a size of 70 to 80 nm, with a core/shell structure of NaYF4:Yb3+:Tm3+/NaYF4 were used in the study. The nanoparticles have an intensive band of anti-Stokes photoluminescence at a wavelength of 800 nm under irradiation with a wavelength of 975 nm (both wavelengths are within the transparency window for biological tissues). The conversion coefficient of the excitation radiation into the anti-Stokes luminescence was 9%. To increase the time during which nanoparticles can circulate in blood flow of small animals, the nanoparticles were covered by a biocompatible amphiphilic polymer shell. As a tumor model we used Lewis epidermoid carcinoma transfected to mice. Results: We were able to obtain stable water colloids of nanoparticles covered with amphiphilic polymer that could preserve their initial size at least for one month. The use of upconversion nanoparticles with a hydrophilic shell made of intermittent maleic anhydride and octadecene co-polymer with subsequent coating with diglycidyl polyethylene glycol ether allowed for reduction of non-specific reaction of nanoparticles with plasma proteins. In its turn, it resulted in an increased time of their circulation in blood flow of small animals for up to 1 hour. With the Lewis lung carcinoma transfected to mice model we demonstrated аn in-life transportation of upconversion nanoparticles into the tumor with a high degree of localization due to a passive EPR effect. The contrast of luminescent signal in the tumor compared to adjacent tissues was at least 70%. The possibility of visualization of upconverted nanoparticles up to 15 mm of biological tissue was shown. Conclusion: The optical imaging techniques with anti-Stokes photoluminescent markers ensure a high contract real-time detection of tumor tissues that allows for their use for intra-operative diagnostics.
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- 2016
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11. INTERACTION BETWEEN DENDRITIC CELLS AND B CELLS DURING IMMUNE RESPONSE TO T CELL-INDEPENDENT ANTIGENS
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D. A. Khochenkov and M. V. Gavrilova
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дендритные клетки ,полисахарид s3 streptococcus pneumoniae ,поливинипирролидон ,т-независимые антигены типа 2 ,индукция иммунного ответа in vitro ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Abstract. Involvement of dendritic cells (DC) into switching of immune response to T-independent type 2 antigens (TI2 antigens) is poorly studied. The present study addressed some interactions between DCs and TI2-type antigens, i.e., with Streptococcus pneumoniae polysaccharide type 3 (S3), and polyvinylpyrrolidone (PVP, mw. of 360 kDa), as well as a role of antigen-loaded DCs for triggering the immune response. Shortterm treatment of DCs with TI-2 antigens induced their activation manifesting as an increase in numbers of CD80- and CD86-positive cells under the cell culture conditions. DCs loaded with TI-2 antigens were then mixed with normal murine splenocytes and cultured in complete RPMI 1640 medium for 4 days. The numbers of antibody- and immunoglobulin-forming cells (AFCs and IFCs, respectively) were determined by ELISPOT assay. Supplementation with TI2-treated DCs induced a 2.0 to 2.7-fold increase in specific immune response. Antigen-induced polyclonal response was enhanced by 40%. These data suggest a direct interaction between DCs and TI-2 antigens, and their presentation to murine splenocytes, thus leading to both specific and polyclonal B cell activation. B-1 cells are shown to play a main role in such response. Separation of loaded DC and splenocytes by semi-permeable membranes caused inhibition of this immune response.
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- 2014
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12. IN VITRO CELLULAR INTERACTIONS DURING IMMUNE RESPONSE TO T CELLINDEPENDENT TYPE 2 ANTIGENS
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M. V. Gavrilova, I. N. Chernyshova, D. A. Khochenkov, and E. V. Sidorova
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b1 cells ,th lymphocytes ,тн2 antigens ,в2 lymphocytes ,dendritic cells ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Abstract. Interactions between B1, B2 and bone marrow dendritic cells (DCs) of CBA mice were investigated in the course of immune response to T-independent antigen type 2 (TI 2 Ag) in model system. Splenocytes of Xid-mice CBA/N were used as “fillers”. It was shown that DCs augmented viability of B1 and B2 lymphocytes. Functional activity of the B1 and B2 lymphocytes was determined by ELISPOT method, as relative numbers of antibody- and immunoglobulin-forming cells (AFCs and IFCs, respectively) detectable after four days in culture. An increased immune response to TI 2 Ag in presence of DCs was observed mainly for B2 subpopulation, containing MZ-B lymphocytes as well. Specific effect of DC upon B2 cell subpopulation was confirmed by detection of direct in vitro interactions of DCs with T and B lymphocytes. DCs cultivated with B2 and T lymphocytes, used as fillers, decreased the numbers of AFCs and IFCs, irrespectively of absence or presence of TI-2 Ags. A decrease in AFC and IFC numbers in the cultures with B1 cells was evident in presence of the Ags only. It was shown that the inhibitory effect of DCs depended on their contact interactions. Separate cultures of B1 and B2 cells with DC under transwell conditions resulted into occurence of AFC and IFC in amounts similar to those generated in vitro without DCs.
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- 2014
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13. Synthesis and anticancer activity of novel 2-alkylthio-4-amino-5-(thiazol-2-YL)pyrimidines
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Yulia Khochenkova, Yulia S. Machkova, Marina A. Troshina, Rovshan E. Gasanov, Elena V. Varakina, D. A. Khochenkov, Olga S. Avdyakova, and Alexander S. Bunev
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Fist ,Chemistry ,Organic Chemistry ,Cancer cell lines ,Combinatorial chemistry - Abstract
A series of new fist-time synthesized of thiazolyl-substituted 4-aminopyrimidine was obtained on the basis of a two-stage process including thioamidation of 4-amino-5-cyanopyrimide followed by one-pot acylation and Hantzsch synthesis in good yield (52–77%). All synthesized compounds showed a good anticancer effect in antiproliferative assay (end point) on three tumor lines A431, A549 и HT-1080
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- 2021
14. Rapamycin synergizes the cytotoxic effects of MEK inhibitor binimetinib and overcomes acquired resistance to therapy in melanoma cell lines in vitro
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Roman Akasov, Anastasia Prokofieva, I. S. Abramov, Nataly V Sholina, Oxana Ryabaya, and D. A. Khochenkov
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0301 basic medicine ,Pharmacology ,Programmed cell death ,Chemistry ,Melanoma ,MEK inhibitor ,Binimetinib ,medicine.disease ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,Oncology ,Apoptosis ,030220 oncology & carcinogenesis ,medicine ,Cancer research ,Pharmacology (medical) ,Viability assay ,Protein kinase B ,PI3K/AKT/mTOR pathway - Abstract
Objective The problem of drug resistance to BRAF-targeted therapy often occurs in melanoma treatment. Activation of PI3K/AKT/mTOR signaling pathway is one of the mechanisms of acquired resistance and a potential target for treatment. In the current research, we investigated that dual inhibition of mTOR and MEK synergistically reduced the viability of melanoma cells in vitro. Methods A combination of rapamycin (a macrolide immunosuppressant, mTOR inhibitor) and binimetinib (an anti-cancer small molecule, selective inhibitor of MEK) was studied using a panel of melanoma cell lines, including patient-derived cells. Results It was found, that combinatorial therapy of rapamycin (250 nM) and binimetinib (2 μM) resulted in 25% of cell viability compared to either rapamycin (85%) or binimetinib alone (50%) for A375 and vemurafenib-resistant Mel IL/R cells. The suppressed activation of mTOR and MEK by combined rapamycin and binimetinib treatment was confirmed using Western blot assay. Cell death occured via the apoptosis pathway; however, the combination treatment significantly increased the apoptosis only for Mel IL/R cells. The enhanced cytotoxic effect was also associated with enhanced cell cycle arrest in the G0/G1 phase. Conclusion In general, we provide the evidence that dual inhibition of mTOR and MEK could be promising for further preclinical investigations.
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- 2021
15. Upconversion Nanoparticles Decorated with Polysialic Acid for Solid Tumors Visualization In Vivo
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Deev Sm, Polina A. Demina, D. A. Khochenkov, Evgeny V. Khaydukov, A. V. Nechaev, N. V. Sholina, Roman Akasov, Alla N. Generalova, and Irina V. Balalaeva
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polysialic acid ,Biocompatibility ,Biophysics ,upconversion nanophosphors ,02 engineering and technology ,Signal-To-Noise Ratio ,Biochemistry ,03 medical and health sciences ,Upconversion nanoparticles ,In vivo ,Cell Line, Tumor ,Humans ,Tissue Distribution ,bioimaging ,surface functionalization ,030304 developmental biology ,0303 health sciences ,Polysialic acid ,Chemistry ,General Chemistry ,General Medicine ,021001 nanoscience & nanotechnology ,Molecular Imaging ,Nanomedicine ,Biochemistry, Biophysics, and Molecular Biology ,Sialic Acids ,Nanoparticles ,Surface modification ,Circulation time ,0210 nano-technology ,Preclinical imaging ,Protein adsorption - Abstract
Abstract Upconversion nanoparticles (UCNPs) are a promising nanoplatform for bioreagent formation for in vivo imaging, which emit UV and blue light under the action of near-infrared radiation, providing deep tissue penetration and maintaining a high signal-to-noise ratio. In the case of solid tumor visualization, the UCNP surface functionalization is required to ensure a long circulation time, biocompatibility, and non-toxicity. The effective UCNP accumulation in the solid tumors is determined by the disturbed architecture of the vascular network and lymphatic drainage. This work demonstrates an approach to the UCNP biofunctionalization with endogenous polysialic acid for in vivo bioreagent formation. Bioreagents possess a low level of nonspecific protein adsorption and macrophage uptake, which allow the prolongation of the circulation time in the bloodstream up to 3 h. This leads to an intense photoluminescent signal in the tumor.
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- 2021
16. THE VALUE OF BASAL EXPRESSION LEVEL OF HEMOXYGENASE-1 FOR SENSITIVITY OF HUMAN MELANOMA CELLS TO OXIDATIVE STRESS IN VITRO
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D. A. Khochenkov, T. A. Sidorova, A. A. Prokofieva, Yu. A. Khochenkova, and E. Sh. Solomko
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0301 basic medicine ,Chemistry ,General Engineering ,HEMOXYGENASE 1 ,medicine.disease_cause ,Molecular biology ,In vitro ,03 medical and health sciences ,Basal (phylogenetics) ,030104 developmental biology ,0302 clinical medicine ,030220 oncology & carcinogenesis ,medicine ,General Earth and Planetary Sciences ,Human melanoma ,Oxidative stress ,General Environmental Science - Abstract
Introduction.The molecular basis of radio- and photodynamic therapy (PDT), the mechanism of action of a number of antitumor chemotherapy drugs is oxidative stress (OS). The enzyme hemoxygenase-I (НO-1), a molecular marker of OS, is a key participant in the system of protection and adaptation of tumor cells under stress.Objective.To find out whether the sensitivity of human melanoma tumor cells to OS depends on the basal and modulator-induced levels of НO-1 expressionMaterial and methods.Human melanoma cell lines were used in the study. The expression of mRNA НO-1 in cells was studied by real-time RT-PCR, the reactive oxygen species content in cells – by flow cytometry and the cytotoxicity of drugs – by MTT assay.Results.According to our data, human melanoma cells have different basal levels of HO-1 transcription: high (3.0–3.5 o. u.) in lines MelIL, MelP, medium (1.5 o. u.) in lines MeWo, MelZ, MelIbr and low (0.5 o. e.) – MelMe, A375).There is no direct correlation between the level of basal cell expression of HO-1 and their sensitivity to the OS inducer – Н2О2. The hemin-induced increase in HO-1 expression in cells is accompanied by doubled resistance to Н2О2. It was found that HO-1 repression in the presence of apigenin was registered in melanoma cells with different basal levels, but sensitization to Н2О2 (2–4 times) was observed only for cells with medium (MeWo) and low (A375) levels of basal HO-1 expression. It was found that the decrease in basal expression of HO-1 induced by apigenin is accompanied by an increase in the reactive oxygen species content in cells.Conclusions.The results of our research allow us to recommend natural flavon apigenin, a modulator of HO-1 expression, for inclusion in the chemotherapy and PDT regimens to increase the effectiveness of human melanoma treatment.
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- 2020
17. Expression of growth factors and tyrosine kinase receptors in the primary tumor and tumor thrombus cells in patients with renal cell carcinoma
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M. I. Volkova, A. S. Olshanskaya, I. V. Tsimafeyeu, N. L. Vashakmadze, Yu. A. Khochenkova, E. Sh. Solomko, S. A. Ashuba, D. A. Khochenkov, and V. B. Matveev
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renal cell carcinoma ,fibroblast growth factor 2 (fgf-2) ,fgfr-1, -2 ,Urology ,030232 urology & nephrology ,vascular endothelial growth factor (vegf-a) ,03 medical and health sciences ,0302 clinical medicine ,vegfr-1, -2 receptors ,Growth factor receptor ,Renal cell carcinoma ,medicine ,Radiology, Nuclear Medicine and imaging ,Thrombus ,platelet-derived growth factor receptors (pdgfr-α, pdgfr-β) ,biology ,business.industry ,medicine.disease ,Primary tumor ,tumor venous thrombosis ,Vascular endothelial growth factor A ,Oncology ,Nephrology ,030220 oncology & carcinogenesis ,embryonic structures ,cardiovascular system ,biology.protein ,Cancer research ,Medicine ,Immunohistochemistry ,Surgery ,business ,Platelet-derived growth factor receptor ,Clear cell - Abstract
Objective: to assess the expression and prognostic value of vascular endothelial growth factor A (VEGF-A), fibroblast growth factor 2 (FGF-2) and their receptors VEGFR-1, -2; FGFR-1, -2, as well as platelet-derived growth factor receptors (PDGFR-α, PDGFR-β) in paired samples of primary tumors and tumor thrombi in renal cell carcinoma (RCC).Materials and methods. Expression of VEGF-A, FGF-2, VEGFR-1, -2; FGFR-1, -2; PDGFR-α, -β was studied in paired surgical samples of primary tumors and tumor thrombi in 25 patients with clear cell RCC pT3a–T4N0–1M0–1 and tumor venous thrombosis by immunohistochemical assay using the appropriate Abcam/Santa Cruz Biotech antibodies from the immunohistochemical staining kit Invitrogen. Expression levels were evaluated by a semi-quantitative method (H-score). The analysis of the correlation between expression levels of VEGF-A, FGF-2, VEGFR-1, -2; FGFR-1, -2; PDGFR-α, -β and RCC characteristics, as well as evaluation of their influence on the outcome of RCC were performed.Results. VEGF-A, FGF-2, as well as VEGFR-1, -2; FGFR-1, -2; PDGFR-α, -β were expressed in the cytoplasm and on the membrane of the primary tumor and tumor thrombus cells in RCC patients. Tumor thrombus cells were characterized by lower expression of VEGFR-1, VEGFR-2, PDGFR-α (p ³2 identified risk factors was 27.3 %, risk factors – 87.5 % (p = 0.004).Conclusion. Tumor thrombus cells in RCC patients expressed VEGF-A, FGF-2, VEGFR-1, -2; FGFR-1, -2; PDGFR-α, -β less active than the cells of the primary tumor. Overexpression of growth factors and tyrosine kinases correlated with RCC Furman grade and tumor venous wall invasion. Overexpression of VEGFR-2 in both primary tumor and thrombus cells in combination with hypoexpression of VEGF-A in the thrombus negatively influenced on OS.
- Published
- 2020
18. In vivo monitoring of vascularization and oxygenation of tumor xenografts using optoacoustic microscopy and diffuse optical spectroscopy
- Author
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K. G. Akhmedzhanova, A. A. Kurnikov, D. A. Khochenkov, Yu. A. Khochenkova, A. M. Glyavina, V. V. Kazakov, A. V. Yudintsev, A. V. Maslennikova, I. V. Turchin, P. V. Subochev, and A. G. Orlova
- Subjects
Article ,Atomic and Molecular Physics, and Optics ,Biotechnology - Abstract
The research is devoted to comparison of the blood vessel structure and the oxygen state of three xenografts: SN-12C, HCT-116 and Colo320. Differences in the vessel formation and the level of oxygenation are revealed by optoacoustic (OA) microscopy and diffuse optical spectroscopy (DOS) respectively. The Colo320 tumor is characterized by the highest values of vessel size and fraction. DOS showed increased content of deoxyhemoglobin that led to reduction of saturation level for Colo320 as compared to other tumors. Immunohistochemical (IHC) analysis for CD31 demonstrates the higher number of vessels in Colo320. The IHC for hypoxia was consistent with DOS results and revealed higher values of the relative hypoxic fraction in Colo320.
- Published
- 2022
19. Synthesis and cytotoxic activity of novel 4-amino-5-cyano-2-sulfonylpyrimidines
- Author
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Alexander S. Bunev, Stepanova Ev, D. A. Khochenkov, Yulia Khochenkova, Rovshan E. Gasanov, and Yulia S. Machkova
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chemistry.chemical_classification ,Isothiouronium ,Sulfide ,010405 organic chemistry ,Chemistry ,General Chemistry ,Cell cycle ,010402 general chemistry ,01 natural sciences ,Combinatorial chemistry ,0104 chemical sciences ,chemistry.chemical_compound ,Apoptosis ,LNCaP ,Cytotoxic T cell ,Moiety ,Malononitrile - Abstract
Novel 4-amino-5-cyano-2-sulfonylpyrimidines were prepared based on three-component cyclization between isothiouronium salts, benzaldehydes and malononitrile, followed by oxidation of the sulfide moiety with Oxone. The cytotoxic activity of the synthesized compounds, as well as the induction of apoptosis, inhibition of the cell cycle and proliferation tests were performed on selected cancer cell lines A431, A549, A375, HCT 116, MCF7, LNCap and SH-SY5Y.
- Published
- 2020
20. Advantages and Possibilities of Fluorescence-Based Methods for the Visualization of Apoptosis and Autophagy in Human Tumor Cells in vitro
- Author
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G. N. Maslyakova, D. A. Mudrak, A. V. Polukonova, Nikita A. Navolokin, Alla B. Bucharskaya, N. V. Polukonova, A. M. Myl’nikov, M. A. Baryshnikova, and D. A. Khochenkov
- Subjects
010302 applied physics ,Programmed cell death ,Muse cell ,biology ,Autophagy ,Acridine orange ,biology.organism_classification ,Apoptotic body ,01 natural sciences ,Atomic and Molecular Physics, and Optics ,Electronic, Optical and Magnetic Materials ,Cell biology ,010309 optics ,HeLa ,chemistry.chemical_compound ,chemistry ,Apoptosis ,0103 physical sciences ,Propidium iodide - Abstract
The possibilities of using fluorescence-based analytical methods and their advantages for visualization and identification of the type of programmed cell death in human tumor cells exposed to flavonoids have been analyzed in experiments in vitro. HeLa cervical cancer cells and A498 kidney carcinoma cells were used as the objects of the study, and exposure to the flavonoid-containing extract of common hedgehyssop (Gratiola officinalis L.) was tested as the experimental treatment. The following fluorescence-based techniques were used: the “live and dead” test with double staining by propidium iodide and acridine orange and double staining with annexin and propidium iodide. Autophagy induction was confirmed by fluorescence-based tests implemented in a Muse cell analyzer with a Muse Autophagy LC3-Antibody Based Kit. The use of double staining with acridine orange and propidium iodide fluorescent dyes in the “live and dead” test and comparison with phase contrast microscopy enables the visualization of apoptotic body and autophagosome formation processes in the cells and can, therefore, be used as a method of screening assessment of the efficiency of various chemotherapy drugs.
- Published
- 2019
21. 1-Imidoyl-1,2,3-benzotriazoles—Novel Reagents for the Synthesis of 1-Aryl-5-trifluoromethylimidazoles
- Author
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E. V. Varakina, D. A. Khochenkov, Alexander S. Bunev, and Alexander S. Peregudov
- Subjects
Trifluoromethyl ,Benzotriazole ,010405 organic chemistry ,Aryl ,Sodium ,Organic Chemistry ,chemistry.chemical_element ,01 natural sciences ,Medicinal chemistry ,0104 chemical sciences ,chemistry.chemical_compound ,chemistry ,Reagent ,Tosylmethyl isocyanide ,Van Leusen reaction - Abstract
1-Imidoylbenzotriazoles [N-aryl-1-(1H-benzotriazol-1-yl)-2,2,2-trifluoroethan-1-imines] reacted with tosylmethyl isocyanide according to van Leusen’s procedure to give difficultly accessible 1-aryl-4-(4-methylbenzenesulfonyl)-5-(trifluoromethyl)-1H-imidazoles in good yields (81–94%). The initial imidoylbenzotriazoles were conveniently synthesized by reaction of sodium benzotriazolide with the corresponding imidoyl chlorides in THF. The reactions were carried out with a wide series of imidoylbenzotriazoles containing various electron-donating and electron-withdrawing substituents in the N-aryl fragment.
- Published
- 2019
22. A versatile platform for bioimaging based on colominic acid-decorated upconversion nanoparticles
- Author
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D. A. Khochenkov, N. V. Sholina, Polina A. Demina, Evgeny V. Khaydukov, Roman Akasov, Alla N. Generalova, Andrei V Nechaev, and Natalia A. Arkharova
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Biomedical Engineering ,Polyethylene glycol ,engineering.material ,Hydrophilization ,Polyethylene Glycols ,Surface coating ,chemistry.chemical_compound ,Adsorption ,chemistry ,Coating ,Covalent bond ,In vivo ,Polysaccharides ,Biophysics ,engineering ,Surface modification ,Animals ,Nanoparticles ,Polyethyleneimine ,General Materials Science - Abstract
Lanthanide-doped upconversion nanoparticles (UCNPs) are promising bioimaging agents that emit light under near infra-red excitation, capable of penetrating deep in biotissues with a high signal-to-noise ratio. Their successful implementation is principally associated with surface functionalization. Here, we report on UCNP surface modification with highly hydrophilic, endogenous, non-toxic, non-immunogenic colominic acid, conferring "stealth" properties. We proposed surface functionalization of UCNPs based on a two-step strategy, which consists of hydrophilization with polyethyleneimine and attachment of colominic acid by electrostatic or covalent bond formation. Analysis revealed that regardless of the nature of the bond, colominic acid acted as a non-cytotoxic UCNP surface coating with low nonspecific blood protein adsorption. UCNP-colominic acid nanocomplexes exhibited low uptake by macrophages in vitro, which plays an active role in inflammatory reactions. We demonstrated the superiority of colominic acid compared to polyethylene glycol coating in terms of the prolonged circulation time in the bloodstream of small animals when injected intravenously. The colominic acid coating made it possible to prolong the UCNP circulation time up to 3 h. This led to the efficient UCNP accumulation in the inflammation site due to microvascular remodeling, accompanied by an enhanced uptake and retention effect. UCNP-assisted imaging of inflammation in the whole-body mode as well as local visualization of blood vessels were acquired in vivo. These collective findings validate the functional significance of UCNP decoration with colominic acid for their application in bioimaging.
- Published
- 2020
23. Nanosized Anti-Stokes Phosphors for Antitumor Drug Delivery and Solid Tumor Theranostics
- Author
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Evgeny V. Khaydukov, Roman Akasov, V V Zasedateleva, Alla N. Generalova, V. Ya. Panchenko, J. Senthil Selvan, Polina A. Demina, N. V. Sholina, and D. A. Khochenkov
- Subjects
Luminescence ,Polymers ,Biophysics ,Nanotechnology ,Phosphor ,Biochemistry ,Polyvinyl alcohol ,Theranostic Nanomedicine ,chemistry.chemical_compound ,Drug Delivery Systems ,Polylactic acid ,Cell Line, Tumor ,Neoplasms ,medicine ,Humans ,Doxorubicin ,Precision Medicine ,Solid tumor ,Antibiotics, Antineoplastic ,General Chemistry ,General Medicine ,Photon upconversion ,Dextran ,chemistry ,Drug delivery ,Nanoparticles ,medicine.drug - Abstract
Theranostics is the direction in modern biomedicine aimed at developing drugs that combine the capabilities of diagnosis and therapy of tumors in one agent. Upconversion nanophosphors (UCNPs) are inorganic crystalline materials that can be used to create a nanoplatform providing diagnostic and therapeutic modalities. They have been proposed as luminescent markers for optical imaging of biological tissue due to their anti-Stokes luminescence, lack of photodegradation and low toxicity. In this article, UCNPs as a theranostic agent for both optical imaging and delivery of anticancer drugs have been offered. To obtain biocompatible nanocomplexes, UCNP surface with a core/shell structure of NaYF4:Yb3+Tm3+/NaYF4 was modified with polylactic acid in the presence of various stabilizers (dextran, polyvinyl alcohol, and poly-N-vinylpyrrolidone). To give the therapeutic modality to the nanocomplex, the antitumor antibiotic doxorubicin was loaded into the polymer shell. The loading efficiency was up to 0.1 mg per 1 mg UCNPs. The toxicity and the intracellular accumulation of nanocomplexes were evaluated in vitro. It was concluded that the modification of UCNPs with polylactic acid provides the transport of doxorubicin, allowing the combination of diagnostic and therapeutic modalities in one agent.
- Published
- 2020
24. The Susceptibility of Human Melanoma Cells to Infection with the Leningrad-16 Vaccine Strain of Measles Virus
- Author
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Denis Vorobyev, Marina Gavrilova, T. V. Nasedkina, Elena Prokofeva, Zverev Vv, D. A. Khochenkov, Yulia Ammour, Oxana Ryabaya, Oxana Svitich, Evgeny Faizuloev, Igor Shohin, and Yulia Shchetinina
- Subjects
0301 basic medicine ,medicine.medical_treatment ,Measles Vaccine ,lcsh:QR1-502 ,Mice, Nude ,Vaccines, Attenuated ,Article ,lcsh:Microbiology ,Measles virus ,03 medical and health sciences ,0302 clinical medicine ,Cancer immunotherapy ,Cell Line, Tumor ,Virology ,medicine ,Animals ,Humans ,Melanoma ,Oncolytic Virotherapy ,Mice, Inbred BALB C ,Attenuated vaccine ,cancer immunotherapy ,biology ,Interferon-stimulated gene ,biology.organism_classification ,medicine.disease ,Xenograft Model Antitumor Assays ,Oncolytic virus ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,Cell killing ,Cell culture ,oncolytic viruses ,030220 oncology & carcinogenesis ,measles virus ,Female - Abstract
Oncolytic viruses, including live attenuated measles virus (MV) vaccine strains, have recently been shown as promising therapeutic agents against human malignancies. In this study, the oncolytic potential of the attenuated vaccine strain Leningrad-16 (L-16) of MV was evaluated in a panel of human metastatic melanoma cell lines. The L-16 measles virus was shown to replicate within melanoma cells mediating direct cell killing of tumor cells, although all melanoma cell lines varied in regard to their ability to respond to L-16 MV infection, as revealed by the different pattern of the Interferon Stimulated Gene expression, cytokine release and mechanisms of cell death. Furthermore, the statistically significant L-16 measles virus related tumor growth inhibition was demonstrated in a melanoma xenograft model. Therefore, L-16 MV represents an appealing oncolytic platform for target delivery of therapeutic genes along with other attenuated measles virus strains.
- Published
- 2020
25. Inactivation of Receptor Tyrosine Kinases Overcomes Resistance to Targeted B-RAF Inhibitors in Melanoma Cell Lines
- Author
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O. O. Ryabaya, A. A. Malysheva, Yu. A. Khochenkova, E. Sh. Solomko, and D. A. Khochenkov
- Subjects
0301 basic medicine ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Structural Biology ,030220 oncology & carcinogenesis ,Biophysics - Published
- 2018
26. ПРЕОДОЛЕНИЕ РЕЗИСТЕНТНОСТИ КЛЕТОЧНЫХ ЛИНИЙ МЕЛАНОМЫ К ТАРГЕТНЫМ ИНГИБИТОРАМ B-RAF ЗА СЧЕТ ИНАКТИВАЦИИ РЕЦЕПТОРНЫХ ТИРОЗИНКИНАЗ, 'Молекулярная биология'
- Author
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E. S. . Solomko, A. A. Malysheva, Y. . A. Khochenkova, D. A. Khochenkov, and Oxana Ryabaya
- Subjects
0301 basic medicine ,biology ,Chemistry ,Cell growth ,Receptor Protein-Tyrosine Kinases ,General Medicine ,Receptor tyrosine kinase ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,030220 oncology & carcinogenesis ,biology.protein ,medicine ,Cancer research ,Phosphorylation ,Signal transduction ,Vemurafenib ,Tyrosine kinase ,Platelet-derived growth factor receptor ,medicine.drug - Abstract
The discovery of B-RAF activating mutations in malignant melanoma cells has led to the development of a number of targeted drugs, which block exclusively the mutant B-RAF protein. Tumor cells often acquire resistance to B-RAF inhibitors via activation of alternative signaling pathways. One of the resistance mechanisms is activation of PDGF, VEGF, c-KIT, and certain other tyrosine kinases. The possibility of overcoming the resistance to the B-RAF inhibitor Vemurafenib by inactivating receptor tyrosine kinases (RTKs) was studied in metastatic melanoma cell lines differing in B-RAF mutations and RTK activity. It was found that RTK inactivation may help to overcome resistance to B-RAF inhibitors via inhibition of tyrosine kinase phosphorylation and a subsequent blocking of the PI3K-AKT-mTOR and MEK-ERK1/2 downstream signaling pathways. The changes eventually mitigated the cell growth and enhanced the Vemurafenib-dependent cell cycle arrest.
- Published
- 2018
27. Nanocurcumin-Loaded UCNPs for Cancer Theranostics: Physicochemical Properties, In Vitro Toxicity, and In Vivo Imaging Studies
- Author
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Anbharasi Lakshmanan, Sergey V. Gudkov, Krishna Bharat Lankamsetty, Ajithkumar Gangadharan, Manonmani Jayaraman, D. A. Khochenkov, N. V. Sholina, Polina A. Demina, Roman Akasov, Alla N. Generalova, Dhiraj K. Sardar, Evgeny V. Khaydukov, and Senthilselvan Jayaraman
- Subjects
theranostics ,Chemistry ,General Chemical Engineering ,technology, industry, and agriculture ,macromolecular substances ,Article ,Photon upconversion ,upconversion nanoparticles ,PLGA ,chemistry.chemical_compound ,nanocurcumin ,In vivo ,herbal drugs ,Zeta potential ,Curcumin ,intravital imaging ,General Materials Science ,Nanocarriers ,QD1-999 ,Preclinical imaging ,Ex vivo ,Nuclear chemistry - Abstract
Formulation of promising anticancer herbal drug curcumin as a nanoscale-sized curcumin (nanocurcumin) improved its delivery to cells and organisms both in vitro and in vivo. We report on coupling nanocurcumin with upconversion nanoparticles (UCNPs) using Poly (lactic-co-glycolic Acid) (PLGA) to endow visualisation in the near-infrared transparency window. Nanocurcumin was prepared by solvent-antisolvent method. NaYF4:Yb,Er (UCNP1) and NaYF4:Yb,Tm (UCNP2) nanoparticles were synthesised by reverse microemulsion method and then functionalized it with PLGA to form UCNP-PLGA nanocarrier followed up by loading with the solvent-antisolvent process synthesized herbal nanocurcumin. The UCNP samples were extensively characterised with XRD, Raman, FTIR, DSC, TGA, UV-VIS-NIR spectrophotometer, Upconversion spectrofluorometer, HRSEM, EDAX and Zeta Potential analyses. UCNP1-PLGA-nanocurcumin exhibited emission at 520, 540, 660 nm and UCNP2-PLGA-nanocurmin showed emission at 480 and 800 nm spectral bands. UCNP-PLGA-nanocurcumin incubated with rat glioblastoma cells demonstrated moderate cytotoxicity, 60–80% cell viability at 0.12–0.02 mg/mL marginally suitable for therapeutic applications. The cytotoxicity of UCNPs evaluated in tumour spheroids models confirmed UCNP-PLGA-nanocurcumin therapeutic potential. As-synthesised curcumin-loaded nanocomplexes were administered in tumour-bearing laboratory animals (Lewis lung cancer model) and showed adequate contrast to enable in vivo and ex vivo study of UCNP-PLGA-nanocurcumin bio distribution in organs, with dominant distribution in the liver and lungs. Our studies demonstrate promise of nanocurcumin-loaded upconversion nanoparticles for theranostics applications.
- Published
- 2021
28. Autophagy inhibitors chloroquine and LY294002 enhance temozolomide cytotoxicity on cutaneous melanoma cell lines in vitro
- Author
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Angelina V. Egorova, Alexander S. Zasedatelev, Evgenia V. Stepanova, Oxana Ryabaya, D. A. Khochenkov, T. V. Nasedkina, M. A. Emelyanova, and Andrey N. Inshakov
- Subjects
0301 basic medicine ,Cancer Research ,Skin Neoplasms ,Morpholines ,Apoptosis ,03 medical and health sciences ,chemistry.chemical_compound ,Chloroquine ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,Autophagy ,Temozolomide ,medicine ,Humans ,Pharmacology (medical) ,LY294002 ,Cytotoxicity ,Melanoma ,neoplasms ,Cell Proliferation ,Pharmacology ,business.industry ,Drug Synergism ,Dacarbazine ,030104 developmental biology ,Oncology ,chemistry ,Chromones ,Cell culture ,Cutaneous melanoma ,Cancer cell ,Cancer research ,business ,medicine.drug - Abstract
Patients with metastatic melanoma are difficult to treat and have a very poor prognosis because of high resistance to therapy. Recent evidence indicates that tumors could overcome death through autophagy, a survival mechanism, which cancer cells use under lack of energy and nutrient deprivation. Melanoma cells have different sensitivity to temozolomide (TMZ) treatment. In this study, we showed that the combination of autophagy inhibitors chloroquine or LY294002 and TMZ induced enhanced cytotoxicity of alkylating agents on human melanoma cell lines. All assays were performed on patient-derived melanoma cell lines. The effectiveness of the combined treatment of TMZ and autophagy inhibitors was determined using an MTT assay. Next, we analyzed the expression mRNA level of Beclin 1, LC3B, and p62/STSQM1 and the relative expression of LC3B protein under combined treatment. Autophagic flux was determined by analysis of colocalization of Lysotracker Red and LC3B puncta. Apoptosis was measured by Annexin V/PI staining. Cell cycle analyses were carried out by flow cytometry. We showed that autophagy inhibition could enhance melanoma cell death combined with TMZ therapy. Chloroquine synergistically enhanced the TMZ-induced growth arrest and increased the G0/G1 population in Mel Z and Mel IL cell lines, but not Mel MTP. The expression analysis showed that autophagy involvement in TMZ enhanced cytotoxicity. Furthermore, LY294002, an early-stage autophagy, and PI3K inhibitor were found to exert similar effects. Both chloroquine and LY294002 improved the cytotoxic effect of TMZ treatment, making this combination applicable as a potent antitumor treatment for metastatic melanoma.
- Published
- 2017
29. Ultraviolet phototoxicity of upconversion nanoparticles illuminated with near-infrared light
- Author
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N. V. Sholina, A. V. Nechaev, Andrei V. Zvyagin, K. E. Mironova, D. A. Khochenkov, Sergey M. Deyev, V. A. Semchishen, V. V. Rocheva, Alla N. Generalova, and Evgeny V. Khaydukov
- Subjects
Materials science ,medicine.medical_treatment ,Photodynamic therapy ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Photochemistry ,medicine.disease_cause ,01 natural sciences ,0104 chemical sciences ,Photoexcitation ,DARPin ,Fluorescence microscope ,medicine ,General Materials Science ,Irradiation ,0210 nano-technology ,mCherry ,Phototoxicity ,Ultraviolet - Abstract
Recently introduced upconversion nanoparticles (UCNPs) have pushed the depth of photodynamic therapy (PDT) treatment to the centimetre range by converting deeply-penetrating near-infrared (NIR) radiation to visible radiation for photoexcitation of PDT drugs. Here we demonstrate that the direct exposure of the cancer tissue to phototoxic ultraviolet radiation generated by NIR-photoexcited UCNPs enabled successful PDT. To this aim, core/shell UCNPs of the formula NaYF4:Yb3+Tm3+/NaYF4 featuring an enhanced band in the ultraviolet UV-A and UV-B spectral bands were rationally designed and synthesised. Coupling UCNPs to the recombinant modules of the Designed Ankyrin Repeat Protein (DARPin) fused to a fluorescent protein mCherry allowed the target delivery of DARPin-mCherry/UCNP to human breast adenocarcinoma SK-BR-3 cells overexpressing HER2/neu receptors, as confirmed by fluorescence microscopy. DARPin-mCherry/UCNPs were demonstrated to be phototoxic to SK-BR-3 cells under 975 nm laser irradiation at a dose of 900 J cm−2 due to the UV photoexcitation of endogenous photosensitizers and concomitant generation of reactive oxygen species. The Lewis lung cancer mouse model was employed to demonstrate the feasibility of PDT using UCNP-mediated UV excitation of endogenous photosensitizers in the tumor tissue at a NIR dose of 1200 J cm−2. This study paves the way for exploring and harnessing UV photoexcitation processes in deep tissues in vivo.
- Published
- 2017
30. Photodynamic therapy of melanoma by blue-light photoactivation of flavin mononucleotide
- Author
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Alexander Erofeev, A. Alova, Petr V. Gorelkin, Roman Akasov, Alla N. Generalova, Evgeny V. Khaydukov, N. V. Sholina, and D. A. Khochenkov
- Subjects
Male ,0301 basic medicine ,Cancer therapy ,Light ,Flavin Mononucleotide ,Science ,medicine.medical_treatment ,Mice, Nude ,Flavin mononucleotide ,Apoptosis ,Photodynamic therapy ,Riboflavin ,Article ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Melanoma ,neoplasms ,Cell Proliferation ,Mice, Inbred BALB C ,Photosensitizing Agents ,Multidisciplinary ,Chemistry ,Cancer ,medicine.disease ,Xenograft Model Antitumor Assays ,030104 developmental biology ,Photochemotherapy ,Cancer research ,Medicine ,Skin cancer ,030217 neurology & neurosurgery - Abstract
Melanoma is one of the most aggressive and lethal form of cancer. Photodynamic therapy (PDT) is a clinically approved technique for cancer treatment, including non-melanoma skin cancer. However, the most of conventional photosensitizers are of low efficacy against melanoma due to the possible dark toxicity at high drug concentrations, melanin pigmentation, and induction of anti-oxidant defense mechanisms. In the current research we propose non-toxic flavin mononucleotide (FMN), which is a water-soluble form of riboflavin (vitamin B2) as a promising agent for photodynamic therapy of melanoma. We demonstrated selective accumulation of FMN in melanoma cells in vivo and in vitro in comparison with keratinocytes and fibroblasts. Blue light irradiation with dose 5 J/cm2 of melanoma cells pre-incubated with FMN led to cell death through apoptosis. Thus, the IC50 values of human melanoma A375, Mel IL, and Mel Z cells were in a range of FMN concentration 10–30 µM that can be achieved in tumor tissue under systemic administration. The efficiency of reactive oxygen species (ROS) generation under FMN blue light irradiation was measured in single melanoma cells by a label-free technique using an electrochemical nanoprobe in a real-time control manner. Melanoma xenograft regression in mice was observed as a result of intravenous injection of FMN followed by blue-light irradiation of tumor site. The inhibition of tumor growth was 85–90% within 50 days after PDT treatment.
- Published
- 2019
31. Upconversion nanoparticles for tumor imaging with near-infrared radiation
- Author
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V. A. Semchishen, V. V. Rocheva, E. V. Stepanova, Victor I. Sokolov, Alla N. Generalova, A. V. Nechaev, Evgeny V. Khaydukov, D. A. Khochenkov, and V. Ya. Panchenko
- Subjects
Tumor imaging ,Materials science ,General Physics and Astronomy ,Nanoparticle ,Lewis lung carcinoma ,02 engineering and technology ,respiratory system ,021001 nanoscience & nanotechnology ,Tumor tissue ,respiratory tract diseases ,03 medical and health sciences ,Upconversion nanoparticles ,0302 clinical medicine ,Optical imaging ,Nuclear magnetic resonance ,In vivo ,030220 oncology & carcinogenesis ,0210 nano-technology ,Near infrared radiation - Abstract
Prospects for using upconversion nanoparticles as markers for tumor optical imaging are discussed. Using a model of epidermoid Lewis lung carcinoma engrafted in mice, luminescent signals from nanoparticles delivered into the tumor tissue are registered in vivo.
- Published
- 2016
32. PEG-modified upconversion nanoparticles for in vivo optical imaging of tumors
- Author
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Vitaly P. Zubov, D. A. Khochenkov, V. A. Semchishen, V. V. Rocheva, Alla N. Generalova, Evgeny V. Khaydukov, Boris N. Chichkov, N. V. Sholina, A. V. Nechaev, and Anastasia Koroleva
- Subjects
Diglycidyl ether ,General Chemical Engineering ,Intercalation (chemistry) ,Quantum yield ,Nanotechnology ,02 engineering and technology ,General Chemistry ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Photochemistry ,01 natural sciences ,0104 chemical sciences ,chemistry.chemical_compound ,chemistry ,PEG ratio ,Surface modification ,0210 nano-technology ,Luminescence ,Ethylene glycol ,Protein adsorption - Abstract
A novel surface modification approach of brightly luminescent upconversion nanoparticles (UCNPs) is reported. Inorganic core@shell UCNPs (core – NaYF4 co-doped with Yb3+ and Tm3+ ions, shell – NaYF4) were modified by intercalation with amphiphilic copolymer poly(maleic anhydride-alt-1-octadecene) followed by cross-linking with poly(ethylene glycol) diglycidyl ether (PEG-DGE). The proposed approach enables preparation of UCNPs with an outmost PEG-containing layer, which provides steric stabilization and low non-specific protein adsorption. Intravenous injection of PEG-functionalized UCNPs into the mice results in extension of the UCNP blood circulation time up to 1 hour. In vivo epi-luminescence imaging of the mouse model with Lewis lung carcinoma is ensured by the high quantum yield of the modified UCNPs and passive targeting associated with efficient UCNP accumulation in solid tumors.
- Published
- 2016
33. Metformin increases antitumor activity of MEK inhibitor binimetinib in 2D and 3D models of human metastatic melanoma cells
- Author
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Evgenia V. Stepanova, Roman Akasov, D. A. Khochenkov, Oxana Ryabaya, M. A. Emelyanova, Elena Markvicheva, Sergey V. Burov, Andrey N. Inshakov, and Anastasia Prokofieva
- Subjects
0301 basic medicine ,Neuroblastoma RAS viral oncogene homolog ,Cyclin D ,MAP Kinase Kinase 2 ,MAP Kinase Kinase 1 ,NRAS ,RM1-950 ,BRAF ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Hypoglycemic Agents ,neoplasms ,Melanoma ,Protein Kinase Inhibitors ,Pharmacology ,biology ,Dose-Response Relationship, Drug ,business.industry ,MEK inhibitor ,Binimetinib ,Drug Synergism ,General Medicine ,medicine.disease ,Metformin ,030104 developmental biology ,chemistry ,Apoptosis ,Drug resistance ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Tumor spheroids ,Benzimidazoles ,Therapeutics. Pharmacology ,Cyclin-dependent kinase 6 ,business ,medicine.drug - Abstract
Melanoma is one of the most aggressive and treatment-resistant tumors that responsible for majority of skin-cancer related deaths. Here we propose a combination of MEK inhibitor binimetinib with metformin as a promising therapy against human melanoma cells in vitro, including BRAF -mutated A375, Mel Z, and Mel IL cells, and NRAS-mutated Mel MTP and Mel Me cells. Additionally, we obtained two close to clinical practice models of melanoma progression. The first one was vemurafenib-resistant Mel IL/R melanoma cells with acquired resistance to BRAF inhibition-targeted therapy, and the second one was tumor spheroids, which are 3D in vitro model of small-size solid tumors in vivo. The cytotoxicity of binimetinib and metformin was synergistic in both 2D and 3D melanoma culture and mediated through apoptotic pathway. The combination reduced the number of melanoma-formed colonies, inhibited cell invasion and migration, and led to G0/G1 cell cycle arrest through cyclin D/CDK4/CDK6 pathway. The mechanism of metformin and binimetinib synergy in melanoma cells was associated with increased activation of p-AMPKα and decreased p-ERK, but not with alterations in p-mTOR. In summary, the combination of metformin and binimetinib resulted in stronger anti-proliferative effects on melanoma cells compared to binimetinib alone, and therefore could be promising for clinical applications.
- Published
- 2018
34. Upconversion nanoparticles with anti-Stokes luminescence as bioimaging agents
- Author
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Evgeniy V. Khaydukov, V. V. Rocheva, Natalia V. Sholina, Roman Akasov, Alla N. Generalova, Polina A. Demina, and D. A. Khochenkov
- Subjects
Lanthanide ,Biocompatible polymers ,Physics ,QC1-999 ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Upconversion nanoparticles ,Luminescent Agents ,0210 nano-technology ,Luminescence ,Temperature mapping - Abstract
Lanthanide-based upconversion nanoparticles attach great attention in theranostics due to their unique physicochemical and optical properties. It is innovative platform possessing peculiar properties for luminescent imaging, temperature mapping, sensing, and therapy. In present work we demonstrate advantages of new luminescent agents based on upconversion nanoparticles and hydrophylic biocompatible polymer.
- Published
- 2018
35. Emerging upconversion nanoparticles for industry and biomedical application
- Author
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I. M. Asharchuk, Alla N. Generalova, V. A. Semchishen, A. G. Savelyev, Anastasia V. Sochilina, K.V. Khaydukov, A. V. Nechaev, V. V. Rocheva, D. A. Khochenkov, and Evgeny V. Khaydukov
- Subjects
Upconversion nanoparticles ,Physics ,QC1-999 ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,0210 nano-technology ,01 natural sciences ,0104 chemical sciences - Abstract
In recent years, the overwhelming majority of the upconversion nanoparticles (UCNPs) prominent applications have originated from their unique luminescent properties. Due to original properties of inorganic UCNPs they attract the interest in numerous fields. We discussed a number of UCNP assisted techniques, such as biomedical imaging, therapy agents, anti-counterfeit labels and 3D printing, showing highly versatile and translatable UCNP photoluminescent nanotechnology for the applications in industry and biomedicine.
- Published
- 2018
36. [Inactivation of Receptor Tyrosine Kinases Overcomes Resistance to Targeted B-RAF Inhibitors in Melanoma Cell Lines]
- Author
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O O, Ryabaya, A A, Malysheva, Yu A, Khochenkova, E Sh, Solomko, and D A, Khochenkov
- Subjects
Mitogen-Activated Protein Kinase 1 ,Proto-Oncogene Proteins B-raf ,Phosphatidylinositol 3-Kinases ,Mitogen-Activated Protein Kinase 3 ,Vemurafenib ,Drug Resistance, Neoplasm ,MAP Kinase Signaling System ,Cell Line, Tumor ,TOR Serine-Threonine Kinases ,Humans ,Receptor Protein-Tyrosine Kinases ,Melanoma ,Proto-Oncogene Proteins c-akt - Abstract
The discovery of B-RAF activating mutations in malignant melanoma cells has led to the development of a number of targeted drugs, which block exclusively the mutant B-RAF protein. Tumor cells often acquire resistance to B-RAF inhibitors via activation of alternative signaling pathways. One of the resistance mechanisms is activation of PDGF, VEGF, c-KIT, and certain other tyrosine kinases. The possibility of overcoming the resistance to the B-RAF inhibitor Vemurafenib by inactivating receptor tyrosine kinases (RTKs) was studied in metastatic melanoma cell lines differing in B-RAF mutations and RTK activity. It was found that RTK inactivation may help to overcome resistance to B-RAF inhibitors via inhibition of tyrosine kinase phosphorylation and a subsequent blocking of the PI3K-AKT-mTOR and MEK-ERK1/2 downstream signaling pathways. The changes eventually mitigated the cell growth and enhanced the Vemurafenib-dependent cell cycle arrest.
- Published
- 2016
37. Controlled modification of hyaluronic acid for photoinduced reactions in tissue engineering
- Author
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N. V. Sholina, Anastasia V. Sochilina, Polina A. Demina, Evgeny V. Khaydukov, D. A. Khochenkov, N.V. Ierusalimsky, Roman Akasov, Alla N. Generalova, and A. G. Savelyev
- Subjects
History ,chemistry.chemical_compound ,Tissue engineering ,Chemistry ,Hyaluronic acid ,Biophysics ,Computer Science Applications ,Education - Published
- 2018
38. Riboflavin photoactivation by upconversion nanoparticles for cancer treatment
- Author
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E. V. Stepanova, Sergey M. Deyev, V. A. Semchishen, Andrei V. Zvyagin, A. V. Nechaev, D. A. Khochenkov, K. E. Mironova, Oleg I. Lebedev, Alla N. Generalova, Evgeny V. Khaydukov, V. Ya. Panchenko, Federal Research Center Krasnoyarsk Scientific Center (FRC), Siberian Branch of the Russian Academy of Sciences (SB RAS), N.N. Blokhin Russian Cancer Research Center, Laboratoire de cristallographie et sciences des matériaux (CRISMAT), École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC), Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (IBCh RAS), Russian Academy of Sciences [Moscow] (RAS), Moscow State Technological University (MSTU Stankin), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche sur les Matériaux Avancés (IRMA), Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), and Macquarie University
- Subjects
CHO cell line ,Riboflavin ,Metal Nanoparticles ,02 engineering and technology ,reactive oxygen metabolite ,Inbred C57BL ,01 natural sciences ,Carcinoma, Lewis Lung ,Fluorides ,Mice ,Lewis carcinoma ,Neoplasms ,Ultraviolet light ,Photosensitizer ,animal ,Yttrium ,Ytterbium ,chemistry.chemical_classification ,Multidisciplinary ,Tumor ,Photosensitizing Agents ,Chemistry ,breast tumor ,C57BL mouse ,Photosensitizing Agent ,[CHIM.MATE]Chemical Sciences/Material chemistry ,021001 nanoscience & nanotechnology ,DBA mouse ,[CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry ,Mice, Inbred DBA ,Thulium ,Female ,0210 nano-technology ,Phototoxicity ,experimental neoplasm ,Cell Survival ,sodium yttriumtetrafluoride ,Drug development ,Breast Neoplasms ,photosensitizing agent ,CHO Cells ,010402 general chemistry ,chemistry ,Article ,Cell Line ,Experimental ,Cricetulus ,In vivo ,Cell Line, Tumor ,[CHIM.CRIS]Chemical Sciences/Cristallography ,Animals ,Humans ,Inbred DBA ,[CHIM]Chemical Sciences ,Irradiation ,drug screening ,human ,procedures ,mouse ,Reactive oxygen species ,metal nanoparticle ,fluoride ,Lewis Lung ,Carcinoma ,tumor cell line ,Neoplasms, Experimental ,Xenograft Model Antitumor Assays ,0104 chemical sciences ,Mice, Inbred C57BL ,Photochemotherapy ,drug effects ,Cancer cell ,Biophysics ,pathology ,Biophotonics ,Reactive Oxygen Species ,metabolism - Abstract
Riboflavin (Rf) is a vitamin and endogenous photosensitizer capable to generate reactive oxygen species (ROS) under UV-blue irradiation and kill cancer cells, which are characterized by the enhanced uptake of Rf. We confirmed its phototoxicity on human breast adenocarcinoma cells SK-BR-3 preincubated with 30-μM Rf and irradiated with ultraviolet light, and proved that such Rf concentrations (60 μM) are attainable in vivo in tumour site by systemic intravascular injection. In order to extend the Rf photosensitization depth in cancer tissue to 6 mm in depth, we purpose-designed core/shell upconversion nanoparticles (UCNPs, NaYF4:Yb3+:Tm3+/NaYF4) capable to convert 2% of the deeply-penetrating excitation at 975 nm to ultraviolet-blue power. This power was expended to photosensitise Rf and kill SK-BR-3 cells preincubated with UCNPs and Rf, where the UCNP-Rf energy transfer was photon-mediated with ~14% Förster process contribution. SK-BR-3 xenograft regression in mice was observed for 50 days, following the Rf-UCNPs peritumoural injection and near-infrared light photodynamic treatment of the lesions.
- Published
- 2016
39. Role of dendritic cells in the immune response to T-independent antigens of type 2
- Author
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D. A. Khochenkov
- Subjects
biology ,ELISPOT ,Biophysics ,Cell Biology ,Acquired immune system ,Biochemistry ,Molecular biology ,B-1 cell ,Immune system ,Antigen ,biology.protein ,Antibody ,Pan-T antigens ,CD80 - Abstract
Dendritic cells (DC) belong to the most effective antigen-presenting cells. Their role in the presentation of thymus-dependent antigens and initiation of primary immune response is well known. At the same time, participation of DC in the immune response to T-independent antigens of type 2 (TI-2 antigens) is poorly explored. In this work, the ability of DC to initiate the immune response to a TI-2 antigen α(1→3) dextran (Dex) is investigated. Mouse bone-marrow-derived DC were generated by culturing the precursors with GM-CSF and then DC were pulsed by TI-2 antigens. The pulse induced DC activation, as was verified by an increase in the number of CD80 and CD86 positive cells. Uptake of FITC-labeled Dex was examined by flow cytometry. At a concentration of FITC-Dex of 100 μg/106 cells, the number of DC binding the antigen (Ag) reached “plateau”. DC charged by TI-2 antigens were mixed with normal mouse splenocytes and cultivated in RPMI-1640 medium for 4 days. The numbers of antibody- and immunoglobulin-forming cells were determined by ELISPOT method. The mixtures of splenocytes and naive DC not charged by the Ag were used as control. It was shown that the increase in the numbers of AFC and IFC under the influence of naive DC did not exceed 20%. On the contrary, the addition of DC pulsed by the Ag increased specific immune response more than twofold. The data obtained point to the direct interactions of DC with TI-2 antigens. Pulsed DC present TI-2 antigens to mouse splenocytes and induce specific and polyclonal B-cell activation, i.e., possess immunostimulating activity.
- Published
- 2010
40. Deep tumor imaging by upconversion nanoparticles
- Author
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Nataly V Sholina, Alla N. Generalova, Polina A. Demina, Eugeny Khaydukov, A. V. Nechaev, and D. A. Khochenkov
- Subjects
010302 applied physics ,chemistry.chemical_classification ,Tumor imaging ,Physics ,QC1-999 ,Nanoparticle ,Nanotechnology ,Polymer ,engineering.material ,01 natural sciences ,Upconversion nanoparticles ,Optical imaging ,chemistry ,Coating ,0103 physical sciences ,PEG ratio ,engineering ,Polymer coating ,010306 general physics - Abstract
In this work are shown the prospects of using upconversion nanoparticles (UCNPs) as markers for contrast optical imaging of a tumor. For using nanoparticles for biomedical purposes is implemented a technique for coating nanoparticles with polymers, such as PEG and PSA. This approach provides low non-specific adsorption, which prolongs the circulation of UCNPs in mouse bearing Lewis Lung Cancer (LLC) up to 10 hours. These properties allow nanoparticles to quickly accumulate in the tumor. Effective delivery of particles with different polymer coatings in the tumor is demonstrated with the help of an epiluminescent imaging system.
- Published
- 2018
41. Biology of dendritic cells
- Author
-
D. A. Khochenkov
- Subjects
CD40 ,biology ,Effector ,medicine.medical_treatment ,Biophysics ,chemical and pharmacologic phenomena ,Cell Biology ,Dendritic cell ,Acquired immune system ,Biochemistry ,Cell biology ,Proinflammatory cytokine ,Immune system ,Cytokine ,Antigen ,Immunology ,biology.protein ,medicine - Abstract
Dendritic cells (DC) play a key role in adaptive immune response. By virtue of their extremely wide distribution and high populational diversity, DC interact with almost all types of immune cells linking innate and adaptive immunity. Due to great diversity of receptors, DC recognize a lot of pathogenic microorganisms and namely DC are responsible for the subsequent immune response. Inflammation triggers maturation of DC, which manifests itself in intracellular rearrangement and in appearance of costimulating molecules (CD40, CD80 and CD86) on DC surface. DC capture and process antigens keeping high amount of immunogenic peptides which are then presented to naive lymphocytes and induce their differentiation into effector cells. Depending on pathogen type and cytokine microenvironment, DC induce polarization of immune responses. In the absence of proinflammatory factors DC induce tolerance. In addition, DC play a crucial role in T-lymphocyte selection and Treg formation. The basic traits of DC biology are reviewed.
- Published
- 2008
42. Antiangiogenic Activity of Alofanib, an Allosteric Inhibitor of Fibroblast Growth Factor Receptor 2
- Author
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Nina Peretolchina, E. Sch. Solomko, D. A. Khochenkov, Evgenia V. Stepanova, and Oxana Ryabaya
- Subjects
Male ,Vascular Endothelial Growth Factor A ,Angiogenesis ,Allosteric regulation ,Mice, Nude ,Angiogenesis Inhibitors ,Vascular endothelial growth inhibitor ,Benzoates ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Mice ,Random Allocation ,Allosteric Regulation ,In vivo ,Human Umbilical Vein Endothelial Cells ,Animals ,Humans ,Receptor, Fibroblast Growth Factor, Type 2 ,Receptor ,Ovarian Neoplasms ,Matrigel ,Sulfonamides ,Alanine ,Neovascularization, Pathologic ,Fibroblast growth factor receptor 2 ,Chemistry ,Heparin ,Triazines ,Endothelial Cells ,General Medicine ,Ligand (biochemistry) ,Xenograft Model Antitumor Assays ,Bevacizumab ,Drug Combinations ,Cancer research ,Female ,Fibroblast Growth Factor 2 ,Proteoglycans ,Collagen ,Laminin - Abstract
Alofanib is a potential allosteric inhibitor of FGFR2 used in oncology. The inhibitor blocks the extracellular part of the receptor and prevents its binding with the ligand. Alofanib suppressed proliferation of endothelial cells, their migration activity, and ability to form vessellike structures in vitro and significantly decreased the number of microvessels in Matrigel implant and in ovarian cancer (SKOV-3) xenograft in vivo. The results indicate that Alofanib can inhibit angiogenesis.
- Published
- 2015
43. Abstract 796: Alofanib, a novel allosteric FGFR2 inhibitor, shows potent antitumor activity in ovarian cancer with FGFR2 expression
- Author
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Evgenia Stepanova, Ilya Tsimafeyeu, Sergei Tjulandin, Daniel Harrison, D. A. Khochenkov, and Mikhail Byakhov
- Subjects
Cancer Research ,Chemotherapy ,Cell growth ,business.industry ,medicine.medical_treatment ,Cancer ,Pharmacology ,medicine.disease ,Carboplatin ,chemistry.chemical_compound ,Breast cancer ,Oncology ,chemistry ,Paclitaxel ,medicine ,Growth inhibition ,Ovarian cancer ,business - Abstract
Alofanib (formerly known as RPT835) is a novel allosteric FGFR2 inhibitor with activity in FGFR2-expressing triple-negative breast cancer [Tjulandin S, et al. San Antonio Breast Cancer Symposium 2014]. Early data suggest that combining FGFR2 inhibitors with platinum-containing cytotoxic agents for the treatment of epithelial ovarian cancer may yield increased antitumor activity [Cole C, et al. Cancer Biol Ther. 2010]. We investigated antitumor activity of alofanib in ovarian cancer in vitro and in vivo. To assess the efficacy of alofanib on FGF-mediated cell proliferation, ovarian cancer (SKOV-3) FGFR2-expressing cells were incubated in a 96-well microculture plate and were treated with serially diluted RPT835. Basic FGF was added at a concentration of 25 ng/ml. Control wells were left untreated. Cell growth inhibition was determined using Promega's Cell Titer-Glo® assay. Immunocompromised mice were used for xenotransplantation of FGFR2 high-expressing ovarian cancer cells (SCOV-3). Seventy animals with measurable tumors were selected on day 10 and randomized into control groups (no treatment or chemotherapy alone (paclitaxel + carboplatin) and treatment groups (alofanib orally or intravenously (different dose levels) in combination with chemotherapy). Measurements of tumor volume (mm3) were performed by digital calipers every 3 days during 30 days after tumor inoculation. Basic FGF significantly increased proliferation of the ovarian cancer cells in untreated control group (P = 0.001). Alofanib treatment resulted in growth inhibition of SKOV-3 cell line in vitro. Treatment of alofanib in combination with paclitaxel/carboplatin demonstrated significant tumor growth delay phenotype in all treatment groups compared to control non-treatment groups. Alofanib exhibited a dose-dependent effect on tumor growth. Daily intravenous regimen of alofanib (total maximum dose per week was 350 mg/kg) demonstrated dramatic effect (inhibiting growth by 80% and by 53% in comparison with vehicle and chemotherapy group alone, respectively (P These results provide strong rationale for evaluation of alofanib in combination with paclitaxel and carboplatin in patients with ovarian cancer. Citation Format: Sergei Tjulandin, Mikhail Byakhov, Evgenia Stepanova, Dmitry Khochenkov, Daniel Harrison, Ilya Tsimafeyeu. Alofanib, a novel allosteric FGFR2 inhibitor, shows potent antitumor activity in ovarian cancer with FGFR2 expression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 796. doi:10.1158/1538-7445.AM2015-796
- Published
- 2015
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