35 results on '"Ruciński, M."'
Search Results
2. EP-2273: Chondrogenic differentiation in vitro results in DDR activation in induced pluripotent stem cells
- Author
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Stelcer, E., primary, Kulcenty, K., additional, Ruciński, M., additional, Jopek, K., additional, Trzeciak, T., additional, Richter, M., additional, and Suchorska, W.M., additional
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- 2018
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3. Ekspresja genu preproghreliny w aspekcie różnorodności wywodzących się z niej peptydów.
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Malendowicz, L., Ruciński, M., and Ziółkowska, A.
- Published
- 2012
4. Stress granule-mediated sequestration of EGR1 mRNAs correlates with lomustine-induced cell death prevention.
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Leśniczak-Staszak M, Pietras P, Ruciński M, Johnston R, Sowiński M, Andrzejewska M, Nowicki M, Gowin E, Lyons SM, Ivanov P, and Szaflarski W
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- Humans, Stress Granules metabolism, Stress Granules genetics, Apoptosis drug effects, Antineoplastic Agents, Alkylating pharmacology, RNA, Messenger metabolism, RNA, Messenger genetics, Early Growth Response Protein 1 metabolism, Early Growth Response Protein 1 genetics, Lomustine pharmacology
- Abstract
Some chemotherapy drugs modulate the formation of stress granules (SGs), which are RNA-containing cytoplasmic foci contributing to stress response pathways. How SGs mechanistically contribute to pro-survival or pro-apoptotic functions must be better defined. The chemotherapy drug lomustine promotes SG formation by activating the stress-sensing eIF2α kinase HRI (encoded by the EIF2AK1 gene). Here, we applied a DNA microarray-based transcriptome analysis to determine the genes modulated by lomustine-induced stress and suggest roles for SGs in this process. We found that the expression of the pro-apoptotic EGR1 gene was specifically regulated in cells upon lomustine treatment. The appearance of EGR1-encoding mRNA in SGs correlated with a decrease in EGR1 mRNA translation. Specifically, EGR1 mRNA was sequestered to SGs upon lomustine treatment, probably preventing its ribosome translation and consequently limiting the degree of apoptosis. Our data support the model where SGs can selectively sequester specific mRNAs in a stress-specific manner, modulate their availability for translation, and thus determine the fate of a stressed cell., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)
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- 2024
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5. Effect of cellular senescence on the response of human peritoneal mesothelial cells to TGF-β.
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Kawka E, Herzog R, Ruciński M, Malińska A, Unterwurzacher M, Sacnun JM, Wagner A, Kowalska K, Jopek K, Kucz-Chrostowska A, Kratochwill K, and Witowski J
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- Humans, Epithelial-Mesenchymal Transition drug effects, Cells, Cultured, Epithelium metabolism, Epithelium drug effects, Signal Transduction drug effects, Gene Expression Profiling, Cellular Senescence drug effects, Transforming Growth Factor beta metabolism, Transforming Growth Factor beta pharmacology, Epithelial Cells metabolism, Epithelial Cells drug effects, Peritoneum cytology, Peritoneum metabolism
- Abstract
Transforming growth factor β (TGF-β) is implicated in both mesothelial-to-mesenchymal transition (MMT) and cellular senescence of human peritoneal mesothelial cells (HPMCs). We previously showed that senescent HPMCs could spontaneously acquire some phenotypic features of MMT, which in young HPMCs were induced by TGF-β. Here, we used electron microscopy, as well as global gene and protein profiling to assess in detail how exposure to TGF-β impacts on young and senescent HPMCs in vitro. We found that TGF-β induced structural changes consistent with MMT in young, but not in senescent HPMCs. Of all genes and proteins identified reliably in HPMCs across all treatments and states, 4,656 targets represented overlapping genes and proteins. Following exposure to TGF-β, 137 proteins and 46 transcripts were significantly changed in young cells, compared to 225 proteins and only 2 transcripts in senescent cells. Identified differences between young and senescent HPMCs were related predominantly to wound healing, integrin-mediated signalling, production of proteases and extracellular matrix components, and cytoskeleton structure. Thus, the response of senescent HPMCs to TGF-β differs or is less pronounced compared to young cells. As a result, the character and magnitude of the postulated contribution of HPMCs to TGF-β-induced peritoneal remodelling may change with cell senescence., (© 2024. The Author(s).)
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- 2024
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6. Estimates of Crop Yield Anomalies for 2022 in Ukraine Based on Copernicus Sentinel-1, Sentinel-3 Satellite Data, and ERA-5 Agrometeorological Indicators.
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Panek-Chwastyk E, Dąbrowska-Zielińska K, Kluczek M, Markowska A, Woźniak E, Bartold M, Ruciński M, Wojtkowski C, Aleksandrowicz S, Gromny E, Lewiński S, Łączyński A, Masiuk S, Zhurbenko O, Trofimchuk T, and Burzykowska A
- Abstract
The study explores the feasibility of adapting the EOStat crop monitoring system, originally designed for monitoring crop growth conditions in Poland, to fulfill the requirements of a similar system in Ukraine. The system utilizes satellite data and agrometeorological information provided by the Copernicus program, which offers these resources free of charge. To predict crop yields, the system uses several factors, such as vegetation condition indices obtained from Sentinel-3 Ocean and Land Color Instrument (OLCI) optical and Sea and Land Surface Temperature Radiometer (SLSTR). It also incorporates climate information, including air temperature, total precipitation, surface radiation, and soil moisture. To identify the best predictors for each administrative unit, the study utilizes a recursive feature elimination method and employs the Extreme Gradient Boosting regressor, a machine learning algorithm, to forecast crop yields. The analysis indicates a noticeable decrease in crop losses in 2022 in certain regions of Ukraine, compared to the previous year (2021) and the 5-year average (2017-2021), specifically for winter crops and maize. Considering the reduction in yield, it is estimated that the decline in production of winter crops in 2022 was up to 20%, while for maize, it was up to 50% compared to the decline in production.
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- 2024
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7. Gene expression profile of hiPSC-derived cells differentiated with growth factors, forskolin and conditioned medium from human adrenocortical cell line.
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Stelcer E, Jopek K, Blatkiewicz M, Olechnowicz A, Kamiński K, Szyszka M, Suchorska WM, and Ruciński M
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- Humans, Culture Media, Conditioned metabolism, Culture Media, Conditioned pharmacology, Intercellular Signaling Peptides and Proteins genetics, Intercellular Signaling Peptides and Proteins metabolism, Intercellular Signaling Peptides and Proteins pharmacology, Transcriptome drug effects, Adrenal Cortex cytology, Adrenal Cortex drug effects, Adrenal Cortex metabolism, Cell Line, Tumor, Adrenal Cortex Neoplasms pathology, Adrenal Cortex Neoplasms genetics, Adrenal Cortex Neoplasms metabolism, Gene Expression Profiling methods, Cell Differentiation drug effects, Induced Pluripotent Stem Cells drug effects, Induced Pluripotent Stem Cells metabolism, Induced Pluripotent Stem Cells cytology, Colforsin pharmacology
- Abstract
Background: Adrenocortical carcinoma (ACC) affects approx. 2 in 1,000,000 individuals in the USA, and is more common in females than males. Adrenocortical carcinoma often presents with severe symptoms, such as abdominal pain, high blood pressure, acne, hair overgrowth, and voice deepening., Objectives: Research on ACC constitutes a large body of published data. There is an increased need for easy access to ACC-derived biological material. Moreover, there are limited numbers of human cell lines available. For this reason, we attempted to differentiate human induced pluripotent stem cells (hiPSCs) into adrenocortical-like cells to establish a new functional cell line., Material and Methods: We conducted a long-term differentiation process (35 and 70 days) in the presence of growth factors (GFs), forskolin and conditioned medium collected from the human adrenal carcinoma (HAC15) cell line. Then, we analyzed the gene expression profile of the differentiated cells., Results: The obtained cells possess features characteristic of all 3 primary germ layers. Interestingly, the differentiated cells demonstrated an extremely high level of gene expression for those involved in endocrine processes, namely glycoprotein hormones, alpha polypeptide (CGA), insulin receptor substrate 4 (IRS4), and pancreatic progenitor cell differentiation and proliferation factor-like protein (PPDPFL)., Conclusions: The results of the study indicate that we obtained progenitors derived from endoderm with some characteristics of pancreatic-like cells. The endodermal derivative differentiation is a very challenging and complicated process; thus, the results presented in this study deserve closer consideration.
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- 2024
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8. Estrogen receptor β affects hypoxia response in colorectal cancer cells.
- Author
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Rawłuszko-Wieczorek AA, Lipowicz J, Nowacka M, Ostrowska K, Pietras P, Blatkiewicz M, Ruciński M, Jagodziński PP, and Nowicki M
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- Humans, Apoptosis genetics, HCT116 Cells, Hypoxia genetics, Colorectal Neoplasms metabolism, Estrogen Receptor beta genetics, Estrogen Receptor beta metabolism
- Abstract
The occurrence of colorectal cancer (CRC) is inversely correlated with estrogen receptor beta (ERβ) presence. Additionally, multiple studies associate low ERβ expression with poorer overall survival of CRC patients. Molecular pathways involved in ERβ - related reduced tumorigenesis include enhanced apoptosis, decreased proliferation, or repression of oncogenes. Moreover, the development of solid tumors, such as CRC, is often associated with an increased tumor mass that results in decreased oxygen partial tension, known as hypoxia, clinically associated with decreased prognosis and therapeutic resistance. Our high-throughput study suggests that ERβ also represses a hypoxic response in CRC cells. We observed a significantly altered transcriptional profile in HCT116 ERβ overexpressing cells that was further stimulated by E2 treatment under hypoxic conditions. The achieved data for downregulation of VEGFA, PDGFA and ANGPTL4 were validated in a time course experiment in DLD-1 cells. In addition, using an ERβ construct with a mutated DNA binding domain we observed that the downregulation of selected genes is dependent on the direct binding of this receptor to regulatory region genes. In addition, we observed that ERβ may affect the expression of the main hypoxia regulator, HIF1A, at the transcriptional and translational levels. In summary, ERβ alters the hypoxic outcome in CRC cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. ARW reports financial support was provided by National Science Centre, Poland., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2024
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9. The response and resistance to drugs in ovarian cancer cell lines in 2D monolayers and 3D spheroids.
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Świerczewska M, Sterzyńska K, Ruciński M, Andrzejewska M, Nowicki M, and Januchowski R
- Abstract
Ovarian cancer is the most common type of gynecologic cancer. One of the leading causes of high mortality is chemoresistance, developed primarily or during treatment. Different mechanisms of drug resistance appear at the cellular and cancer tissue organization levels. We examined the differences in response to the cytotoxic drugs CIS, MTX, DOX, VIN, PAC, and TOP using 2D (two-dimensional) and 3D (three-dimensional) culture methods. We tested the drug-sensitive ovarian cancer cell line W1 and established resistant cell lines to appropriate cytotoxic drugs. The following qualitative and quantitative methods were used to assess: 1) morphology - inverted microscope and hematoxylin & eosin staining; 2) viability - MTT assay; 3) gene expression - a quantitative polymerase chain reaction; 4) identification of proteins - immunohistochemistry, and immunofluorescence. Our results indicate that the drug-sensitive and drug-resistant cells cultured in 3D conditions exhibit stronger resistance than the cells cultured in 2D conditions. A traditional 2D model shows that drug resistance of cancer cells is caused mainly by changes in the expression of genes encoding ATP-binding cassette transporter proteins, components of the extracellular matrix, "new" established genes related to drug resistance in ovarian cancer cell lines, and universal marker of cancer stem cells. Whereas in a 3D model, the drug resistance in spheroids can be related to other mechanisms such as the structure of the spheroid (dense or loose), the cell type (necrotic, quiescent, proliferating cells), drug concentrations or drug diffusion into the dense cellular/ECM structure., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest including employment, consultancies, stock ownership, honoraria, paid expert testimony, patent applications/registrations, and grants or other funding., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)
- Published
- 2023
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10. Impact of Classic Adrenal Secretagogues on mRNA Levels of Urotensin II and Its Receptor in Adrenal Gland of Rats.
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Jopek K, Tyczewska M, Szyszka M, Blatkiewicz M, Jopek M, Malendowicz LK, and Ruciński M
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- Animals, Female, Humans, Male, Rats, Adrenal Glands, Adrenocorticotropic Hormone, RNA, Messenger genetics, Receptors, G-Protein-Coupled drug effects, Receptors, G-Protein-Coupled genetics, Secretagogues, Urotensins drug effects, Urotensins genetics
- Abstract
Urotensin 2 (Uts2) is a biologically active peptide involved in the regulation of a variety of physiological and pathophysiological processes. In both the human and rat adrenal gland, the expressions of the Uts2 gene and its receptor (Uts2r) have been described. This paper focuses on the description of the hormonal control of the mRNA levels of urotensin II and its receptor in the adrenal gland of the rat, both in vitro and in vivo. The initial in vitro experiments were carried out on freshly isolated rat adrenocortical cells and their primary culture. The obtained results indicated a stimulating PKA-independent effect of ACTH on the Uts2 mRNA level in the tested cells, with no changes in the Uts2r transcript. Subsequent in vivo experiments showed that ACTH-induced adrenal growth was accompanied by an elevated level of the Uts2 mRNA, with unchanged expression of Uts2r . In the other types of in vivo gland growth studied, enucleation-induced adrenal regeneration and compensatory growth of the gland, the mRNA levels of the studied genes showed no significant differences. The only exception was hemiadrenalectomy, which led to a significant increase in Uts2 mRNA expression level 24 h after surgery. In 12-week-old rats of both sexes, gonadectomy led to a significant increase in the level of Uts2 mRNA in the adrenal gland, an effect that was prevented by sex hormones' replacement. No changes in Uts2r transcript levels were observed under these conditions. Thus, this study suggests that the regulation of Uts2 and Uts2r mRNA levels differs significantly in the rat adrenal gland. While Uts2 transcript levels appear to be mainly dependent on ACTH action, Uts2r mRNA levels are not under the control of this hormone.
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- 2023
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11. Apoptosis Related Human Wharton's Jelly-Derived Stem Cells Differentiation into Osteoblasts, Chondrocytes, Adipocytes and Neural-like Cells-Complete Transcriptomic Assays.
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Stefańska K, Nemcova L, Blatkiewicz M, Pieńkowski W, Ruciński M, Zabel M, Mozdziak P, Podhorska-Okołów M, Dzięgiel P, and Kempisty B
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- Humans, Transcriptome, Chondrocytes, Cell Differentiation genetics, Adipocytes, Apoptosis genetics, Osteoblasts, Cells, Cultured, Nerve Tissue Proteins, Wharton Jelly, Mesenchymal Stem Cells
- Abstract
Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) exhibit multilineage differentiation potential, adhere to plastic, and express a specific set of surface markers-CD105, CD73, CD90. Although there are relatively well-established differentiation protocols for WJ-MSCs, the exact molecular mechanisms involved in their in vitro long-term culture and differentiation remain to be elucidated. In this study, the cells were isolated from Wharton's jelly of umbilical cords obtained from healthy full-term deliveries, cultivated in vitro, and differentiated towards osteogenic, chondrogenic, adipogenic and neurogenic lineages. RNA samples were isolated after the differentiation regimen and analyzed using an RNA sequencing (RNAseq) assay, which led to the identification of differentially expressed genes belonging to apoptosis-related ontological groups. ZBTB16 and FOXO1 were upregulated in all differentiated groups as compared to controls, while TGFA was downregulated in all groups. In addition, several possible novel marker genes associated with the differentiation of WJ-MSCs were identified (e.g., SEPTIN4 , ITPR1 , CNR1 , BEX2 , CD14 , EDNRB ). The results of this study provide an insight into the molecular mechanisms involved in the long-term culture in vitro and four-lineage differentiation of WJ-MSCs, which is crucial to utilize WJ-MSCs in regenerative medicine.
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- 2023
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12. The Enhanced Expression of ZWILCH Predicts Poor Survival of Adrenocortical Carcinoma Patients.
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Blatkiewicz M, Kamiński K, Szyszka M, Al-Shakarchi Z, Olechnowicz A, Stelcer E, Komarowska H, Tyczewska M, Klimont A, Karczewski M, Wierzbicki T, Mikołajczyk-Stecyna J, Ruchała M, Malendowicz LK, and Ruciński M
- Abstract
Zwilch kinetochore protein (ZWILCH) plays a key role in proper cell proliferation. The upregulation of the ZWILCH gene was observed in many types of cancers, but the association of ZWILCH with adrenocortical carcinoma (ACC) was not investigated so far. The main aim of the presented study was to verify if the enhanced level of the ZWILCH gene can be used as a diagnostic marker for ACC development and progression, as well as a predictor of survival time for ACC patients. The performed analyses included investigation of the ZWILCH expression profile in tumors with publicly available TCGA (The Cancer Genome Atlas) datasets and transcriptomic data from the Gene Expression Omnibus (GEO) database, as well as, in human biological samples of normal adrenal, adrenocortical carcinoma and in commercially available tissue microarrays. The findings demonstrate statistically significant higher ZWILCH gene expression in ACC tissue in comparison with normal adrenal glands. Furthermore, there is a strong correlation between ZWILCH upregulation and tumor mitotic rate and the probability of patient survival. The enhanced ZWILCH level is also connected with the activation of genes involved in cell proliferation and the inhibition of genes related to the immune system. This work contributes to a better understanding of the role of ZWILCH as an ACC biomarker and diagnostic tool.
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- 2023
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13. Effects of Galp and alarin peptides on HPA axis gene expression and adrenal function: In vivo experiments.
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Tyczewska M, Szyszka M, Jopek K, and Ruciński M
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- Adrenal Glands metabolism, Animals, Cholesterol Side-Chain Cleavage Enzyme genetics, Cholesterol Side-Chain Cleavage Enzyme metabolism, Gene Expression, Mice, Rats, Galanin-Like Peptide genetics, Galanin-Like Peptide metabolism, Hypothalamo-Hypophyseal System metabolism, Pituitary-Adrenal System metabolism
- Abstract
Background: Many experimental data indicate interactions between peptides involved in the control of food intake, energy homeostasis and adrenocortical hormone release. Glucocorticoids stimulate or inhibit the secretion of orexigenic and anorexigenic peptides, which in turn are involved in the regulation of adrenal growth, structure and function. Galanin-like peptide (Galp) and alarin (Ala) are involved in the regulation of food intake. Galp and Ala mRNAs have already been shown to be present in the arcuate nucleus (ARC) of the hypothalamus in both rats and mice., Objectives: To investigate the expression of Ala, Galp and their receptors in the hypothalamus and pituitary and adrenal glands of the rat hypothalamic-pituitary-adrenal (HPA) axis after intraperitoneal administration of peptides in vivo., Material and Methods: Experimental in vivo models were used: acute and long-term exposure to peptides., Results: The expression of Galp, Ala, their receptors, and steroidogenesis enzymes was analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Statistically significant expression changes were found in the hypothalamus and pituitary after 1-hour exposure to the peptides, such as a decrease in corticotropin-releasing hormone (CRH) expression after Ala, Galp and adrenocorticotropic hormone (ACTH) administration, and a decrease in the expression of receptors for galanin (Gal) (Galr1 and Galr2). In the pituitary, there was a statistically significant increase in the expression of Ala, Galr1, Galr2, and Galr3 receptors 1 h after Galp administration. In the adrenal glands, only a statistically significant decrease in Galr2 expression was observed after 1 h of Ala 0.5 administration. The mRNA expression of steroidogenesis enzymes also changed: for example, the expression of cholesterol desmolase increased 24 h after Ala peptide administration., Conclusions: The results indicate that the peptides tested under in vivo conditions can alter the expression of the peptides tested, as well as of Galp, Ala and Gal receptors and steroidogenesis enzymes - Cyp11a1 (cholesterol desmolase), Cyp11b1 (11β-hydroxylase) and Cyp11b2 (aldosterone synthase).
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- 2022
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14. Identification of the Transcriptional Biomarkers Panel Linked to Pathological Remodelling of the Eye Tissues in Various HD Mouse Models.
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Mazur-Michałek I, Ruciński M, Sowiński M, Pietras P, Leśniczak-Staszak M, Szaflarski W, Isalan M, and Mielcarek M
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- Animals, Biomarkers, Disease Models, Animal, Eye Proteins, Membrane Proteins, Mice, Mutant Proteins, Huntington Disease metabolism, Neurodegenerative Diseases
- Abstract
Ocular abnormalities are becoming associated with a spectrum of pathological events in various neurodegenerative diseases. Huntington's disease (HD) is just such an example of a fatal neurological disorder, where mutated genes (CAG trinucleotide expansions in the Huntingtin gene) have widespread expression, leading to the production of mutant Huntingtin (mHTT) protein. It is well known that mutant HTT protein is prone to form toxic aggregates, which are a typical pathological feature, along with global transcriptome alterations. In this study, we employed well-established quantitative methods such as Affymetrix arrays and quantitative PCR (qPCR) to identify a set of transcriptional biomarkers that will track HD progression in three well-established mouse models: R6/2, R6/1, and Hdh Q150. Our array analysis revealed significantly deregulated networks that are related to visual processes and muscle contractions. Furthermore, our targeted quantitative analysis identified a panel of biomarkers with some being dysregulated even at the presymptomatic stage of the disease, e.g., Opn1mw , Opn1sw , and Pfkfb2 . Some of the deregulated genes identified in this study have been linked to other genetic ocular disorders such as: GNAT2, a source of achromatopsia, and REEP6, linked to Retinitis pigmentosa. It may thus be a useful platform for preclinical evaluations of therapeutic interventions.
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- 2022
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15. Biological response of adrenal carcinoma and melanoma cells to mitotane treatment.
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Stelcer E, Komarowska H, Jopek K, Żok A, Iżycki D, Malińska A, Szczepaniak B, Komekbai Z, Karczewski M, Wierzbicki T, Suchorska WM, Ruchała M, and Ruciński M
- Abstract
A previous case report described an adrenal incidentaloma initially misdiagnosed as adrenocortical carcinoma (ACC), which was treated with mitotane. The final diagnosis was metastatic melanoma of unknown primary origin. However, the patient developed rapid disease progression after mitotane withdrawal, suggesting a protective role for mitotane in a non-adrenal-derived tumor. The aim of the present study was to determine the biological response of primary melanoma cells obtained from that patient, and that of other established melanoma and ACC cell lines, to mitotane treatment using a proliferation assay, flow cytometry, quantitative PCR and microarrays. Although mitotane inhibited the proliferation of both ACC and melanoma cells, its role in melanoma treatment appears to be limited. Flow cytometry analysis and transcriptomic studies indicated that the ACC cell line was highly responsive to mitotane treatment, while the primary melanoma cells showed a moderate response in vitro . Mitotane modified the activity of several key biological processes, including 'mitotic nuclear division', 'DNA repair', 'angiogenesis' and 'negative regulation of ERK1 and ERK2 cascade'. Mitotane administration led to elevated levels of DNA double-strand breaks, necrosis and apoptosis. The present study provides a comprehensive insight into the biological response of mitotane-treated cells at the molecular level. Notably, the present findings offer new knowledge on the effects of mitotane on ACC and melanoma cells., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Stelcer et al.)
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- 2022
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16. Extracellular Nampt (eNampt/visfatin/PBEF) directly and indirectly stimulates ACTH and CCL2 protein secretion from isolated rat corticotropes.
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Celichowski P, Jopek K, Szyszka M, Milecka P, Tyczewska M, Sakhanova S, Szaflarski W, Malendowicz LK, and Ruciński M
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- Animals, Cell Communication, Chemokine CCL2, Cytokines, Interleukin-6, Mice, Rats, Adrenocorticotropic Hormone, Nicotinamide Phosphoribosyltransferase
- Abstract
Background: Nicotinamide phosphoribosyltransferase (Nampt/visfatin/PBEF) acts both as an enzyme in the nicotinamide adenine dinucleotide (NAD) synthesis pathway as well as an extracellular hormone (eNampt). Among its effects, eNampt exerts potent pro-inflammatory effects. We have recently shown that, in rats, eNampt stimulates corticosterone secretion by acting through the pituitary rather than the hypothalamus., Objectives: To investigate the mechanism of action of eNampt on the secretion of adrenocorticotropic hormone (ACTH) and chemokine (C-C motif) ligand 2 (CCL2), which are cytokines secreted by pituitary neuroendocrine tumors., Material and Methods: The research was carried out on the AtT-20 murine cell line, primary rat pituitary cell culture, isolated pituitary corticotropes, and in vivo. The effects of the performed experiments were examined using the following methods: gene expression profiling using microarrays, quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA)., Results: The results suggest that eNampt stimulates ACTH secretion from rat corticotropes both directly and indirectly. Indirect action most likely occurs through interleukin (IL)-6 secreted by folliculostellate cells of the pituitary gland. In isolated ACTH cells of the rat pituitary gland, eNampt stimulates the expression of genes involved in the immune response. Among them, the protein encoded by the CCL2 gene seems to also be involved in the regulation of corticotropin-releasing hormone (CRH)-dependent metabolism. Unlike rat corticotropes, murine AtT-20 corticotropic cells do not react to either eNampt or Fk866 (the inhibitor of Nampt enzymatic action)., Conclusions: The eNampt stimulates the secretion of ACTH from rat corticotropes indirectly and directly, likely by stimulating IL-6 secretion from folliculostellate cells of the pituitary gland. This effect was not observed in the AtT-20 corticotropic cell cancer cell line.
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- 2021
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17. Changes in total and acylated ghrelin levels during mitotane treatment in patients with adrenocortical carcinoma.
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Komarowska H, Ruciński M, Fichna M, Bromińska B, Iżycki D, Czarnywojtek A, and Ruchała M
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- Adrenal Cortex Neoplasms blood, Adrenocortical Carcinoma blood, Adrenocorticotropic Hormone blood, Adult, Case-Control Studies, Female, Humans, Male, Middle Aged, Adrenal Cortex Neoplasms drug therapy, Adrenocortical Carcinoma drug therapy, Antineoplastic Agents, Hormonal therapeutic use, Ghrelin blood, Mitotane therapeutic use
- Abstract
Introduction: Adrenocortical carcinoma (ACC) is a highly aggressive cancer with poor prognosis. Mitotane is the only approved drug for ACC treatment. Tolerability and efficacy of mitotane is variable. There is evidence that ghrelin may affect cancer development and the occurrence of side effects., Objectives: We examined the differences in plasma ghrelin concentrations between patients with benign adrenal tumors and adrenal carcinoma. We also investigated the effect of mitotane treatment on circulating plasma ghrelin levels in patients with ACC. Additionally, we assessed the relationship between ghrelin concentrations, mitotane levels, and side effects of mitotane treatment., Patients and Methods: We enrolled 26 patients with ACC and 42 controls with adrenocortical adenoma (ACA). Clinical and histopathologic features, hormonal secretion pattern, and plasma acylated and total ghrelin levels were measured in every patient. Serum mitotane levels, body mass index, and side effects of mitotane treatment were estimated every 3 to 12 weeks during follow‑up in patients with ACC., Results: There was no significant difference in total and acylated ghrelin concentrations between ACC and ACA groups before mitotane introduction in ACC. We observed that during mitotane treatment, both total and acylated ghrelin levels became elevated in ACC compared with ACA. A positive correlation was found between circulating mitotane levels and acylated ghrelin as well as the ratio of acylated to total ghrelin levels in all patients treated with mitotane. Higher ghrelin levels were associated with increased risk of side effects., Conclusions: Plasma ghrelin levels are changed during mitotane treatment. These changes may be connected with side effects of mitotane.
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- 2019
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18. Expression profile of Galp, alarin and their receptors in rat adrenal gland.
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Tyczewska M, Milecka P, Szyszka M, Celichowski P, Jopek K, Komarowska H, Malendowicz LK, and Ruciński M
- Subjects
- Animals, Female, Galanin-Like Peptide metabolism, Hypothalamo-Hypophyseal System, Male, Mice, Oligonucleotide Array Sequence Analysis, Pituitary-Adrenal System, Rats, Real-Time Polymerase Chain Reaction, Adrenal Glands metabolism, Galanin-Like Peptide genetics, Hypothalamus metabolism, Pituitary Gland metabolism
- Abstract
Background: Galanin-like peptide (Galp) and alarin (Ala) are 2 new members of the galanin peptide family. Galanin (Gal), the "parental" peptide of the entire family, is known to regulate numerous physiological processes, including energy and osmotic homeostasis, reproduction, food intake, and secretion of adrenocortical hormones. Galp and Ala are known to regulate food intake. In the rat, Galp mRNA has been found in the brain, exclusively in the hypothalamic arcuate nucleus (ARC) and median eminence, which are involved in the regulation of energy homeostasis. Alarin-like immunoreactivity is present in the locus coeruleus (LC) and the ARC of rats and mice., Objectives: The aim of the study was to investigate the expression of Ala, Galp and their receptors in the organs of the hypothalamo-pituitary-adrenal (HPA) axis of the rat., Material and Methods: The expression of the examined genes was measured in different models of adrenal growth of the rat in vivo (postnatal ontogenesis, compensatory adrenal growth, adrenocortical regeneration, adrenocorticotropic hormone (ACTH) administration). The expression was evaluated using the Affymetrix® microarray system or quantitative polymerase chain reaction (qPCR)., Results: The expression of Ala gene was observed in each organ of the HPA axis (the hypothalamus, hypophysis and adrenal gland). The elevated level of expression of this gene was observed in the pituitary of 2-day rats, while very low levels of Ala mRNA were observed in the adrenals. Galp mRNA expression was observed only in the hypothalamus and the hypophysis during postnatal ontogenesis. The expression of Gal receptors was demonstrated in the hypothalamus, the hypophysis and the adrenal gland. In different compartments of the adrenal glands of adult, intact male and female rats, the expression of Ala, Galp and galanin receptor 1 (Galr1) genes was negligible, but the expression of galanin receptor 2 (Galr2), galanin receptor 3 (Galr3) and neurotrophic receptor tyrosine kinase 2 (Ntrk2) genes was noticeable., Conclusions: The examined genes showed different expression levels within the studied HPA axis; some of them were neither expressed in the hypothalamus or the pituitary gland, nor in the adrenal gland.
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- 2019
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19. Nicotinamide phosphoribosyltransferase and the hypothalamic-pituitary-adrenal axis of the rat.
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Celichowski P, Jopek K, Milecka P, Szyszka M, Tyczewska M, Malendowicz LK, and Ruciński M
- Subjects
- Adrenal Cortex cytology, Adrenal Cortex metabolism, Adrenocorticotropic Hormone blood, Adrenocorticotropic Hormone metabolism, Adrenocorticotropic Hormone pharmacology, Aldosterone blood, Aldosterone metabolism, Animals, Cells, Cultured, Corticosterone blood, Corticosterone metabolism, Hypothalamo-Hypophyseal System drug effects, Male, Pituitary-Adrenal System drug effects, Rats, Hypothalamo-Hypophyseal System metabolism, Nicotinamide Phosphoribosyltransferase metabolism, Pituitary-Adrenal System metabolism
- Abstract
Nicotinamide phosphoribosyltransferase (Nampt), also termed visfatin, catalyses the rate‑limiting step in the nicotinamide adenine dinucleotide (NAD) salvage pathway. In addition to its intracellular function (iNampt), extracellular Nampt (eNampt) also affects numerous intracellular signalling pathways. The current study investigated the role of Nampt in the regulation of the hypothalamic‑pituitary‑adrenal (HPA) axis in rats. At 1 h after intraperitoneal administration of eNampt (4 µg/100 g) in adult male rats, serum adrenocorticotropic hormone(ACTH) and aldosterone levels remained unchanged, while corticosterone levels were notably elevated compared with the control group, as determined by ELISA. The results of reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) demonstrated that, in the hypothalami of eNampt‑treated rats, the mRNA expression levels of Fos proto‑oncogene, which is also termed c‑Fos, were not significantly different compared with the control group; however, the mRNA expression levels of proopiomelanocortin (POMC) were markedly increased in the pituitary gland of eNampt‑treated rats compared with the control group. Furthermore, in hypothalamic explants, ELISA results demonstrated that the addition of the eNampt protein exhibited no effect on corticotropin‑releasing hormone (CRH) release into the incubation medium and prevented potassium ion‑induced CRH release. Additionally, the eNampt‑induced increase in ACTH output by pituitary gland explants was not statistically significant, compared with the control group. However, RT‑qPCR indicated that exposure of pituitary gland explants to eNampt and CRH increased the levels of POMC mRNA expression; the effect of eNampt, but not CRH, was inhibited by FK866, which is a specific Nampt inhibitor. In primary rat adrenocortical cell cultures, eNampt exhibited no effect on basal aldosterone or corticosterone secretion, while increases in aldosterone and corticosterone levels in response to ACTH were retained. To assess the potential role of iNampt in the regulation of adrenal steroidogenesis, experiments involving a specific Nampt inhibitor, FK866, were performed. Exposure of cultured cells to FK866 notably lowered basal aldosterone and corticosterone output compared with the control group, and completely eliminated the response of cultured cells to ACTH. The results of the present study indicated that the injected eNampt may have increased the corticosterone serum levels by acting at the pituitary level. In addition, iNampt may exert a tonic stimulating effect on the secretion of aldosterone and corticosterone from rat adrenocortical cells, as normal iNampt levels were required to retain the response of cultured rat adrenocortical cells to ACTH. Thus, these data suggest an important physiological role of both iNampt and eNampt in the regulation of the HPA axis activity in the rat.
- Published
- 2018
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20. Microarray-based detection and expression analysis of new genes associated with drug resistance in ovarian cancer cell lines.
- Author
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Januchowski R, Sterzyńska K, Zawierucha P, Ruciński M, Świerczewska M, Partyka M, Bednarek-Rajewska K, Brązert M, Nowicki M, Zabel M, and Klejewski A
- Subjects
- Antineoplastic Agents pharmacology, Cell Line, Tumor, Computational Biology methods, Female, Gene Expression Profiling methods, Gene Expression Regulation, Neoplastic drug effects, Humans, Immunohistochemistry, Ovarian Neoplasms metabolism, Drug Resistance, Neoplasm genetics, Ovarian Neoplasms genetics
- Abstract
Purpose: The present study is to discover a new genes associated with drug resistance development in ovarian cancer., Methods: We used microarray analysis to determine alterations in the level of expression of genes in cisplatin- (CisPt), doxorubicin- (Dox), topotecan- (Top), and paclitaxel- (Pac) resistant variants of W1 and A2780 ovarian cancer cell lines. Immunohistochemistry assay was used to determine protein expression in ovarian cancer patients., Results: We observed alterations in the expression of 22 genes that were common to all three cell lines that were resistant to the same cytostatic drug. The level of expression of 13 genes was upregulated and that of nine genes was downregulated. In the CisPt-resistant cell line, we observed downregulated expression of ABCC6, BST2, ERAP2 and MCTP1; in the Pac-resistant cell line, we observe upregulated expression of ABCB1, EPHA7 and RUNDC3B and downregulated expression of LIPG, MCTP1, NSBP1, PCDH9, PTPRK and SEMA3A. The expression levels of three genes, ABCB1, ABCB4 and IFI16, were upregulated in the Dox-resistant cell lines. In the Top-resistant cell lines, we observed increased expression levels of ABCG2, HERC5, IFIH1, MYOT, S100A3, SAMD4A, SPP1 and TGFBI and decreased expression levels of MCTP1 and PTPRK. The expression of EPHA7, IFI16, SPP1 and TGFBI was confirmed at protein level in analyzed ovarian cancer patients.., Conclusions: The expression profiles of the investigated cell lines indicated that new candidate genes are related to the development of resistance to the cytostatic drugs that are used in first- and second-line chemotherapy of ovarian cancer.
- Published
- 2017
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21. 3'-hydroxy-3,4,5,4'-tetramethoxystilbene, the metabolite of resveratrol analogue DMU-212, inhibits ovarian cancer cell growth in vitro and in a mice xenograft model.
- Author
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Piotrowska-Kempisty H, Ruciński M, Borys S, Kucińska M, Kaczmarek M, Zawierucha P, Wierzchowski M, Łażewski D, Murias M, and Jodynis-Liebert J
- Subjects
- Animals, Antineoplastic Agents pharmacology, Apoptosis drug effects, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Female, Genome, Human, Humans, Metabolome, Mice, Mice, SCID, Oligonucleotide Array Sequence Analysis, Real-Time Polymerase Chain Reaction, Resveratrol, Signal Transduction drug effects, Tumor Burden drug effects, Tumor Suppressor Protein p53 metabolism, Ovarian Neoplasms pathology, Stilbenes chemistry, Stilbenes metabolism, Stilbenes pharmacology, Xenograft Model Antitumor Assays
- Abstract
In screening studies, the cytotoxic activity of four metabolites of resveratrol analogue 3,4,5,4'-tetramethoxystilbene (DMU-212) against A-2780 and SKOV-3 ovarian cancer cells was investigated. The most active metabolite, 3'-hydroxy-3,4,5,4'-tetramethoxystilbene (DMU-214), was chosen for further studies. The cytotoxicity of DMU-214 was shown to be higher than that of the parent compound, DMU-212, in both cell lines tested. Since DMU-212 was supposed to undergo metabolic activation through its conversion to DMU-214, an attempt was made to elucidate the mechanism of its anti-proliferative activity. We found that in SKOV-3 cells lacking p53, DMU-214 induced receptor-mediated apoptosis. In A-2780 cell line with expression of wild-type p53, DMU-214 modulated the expression pattern of p53-target genes driving intrinsic and extrinsic apoptosis pathways, as well as DNA repair and damage prevention. Regardless of the up-regulation of p48, p53R2, sestrins and Gaad45 genes involved in cancer cell DNA repair, we demonstrated the stronger anti-proliferative and pro-apoptotic effects of DMU-214 in A-2780 cells when compared to those in SKOV-3. Hence we verified DMU-214 activity in the xenograft model using SCID mice injected with A-2780 cells. The strong anti-proliferative activity of DMU-214 in the in vivo model allowed to suggest the tested compound as a potential therapeutic in ovarian cancer treatment.
- Published
- 2016
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22. Microarray-based detection and expression analysis of extracellular matrix proteins in drug‑resistant ovarian cancer cell lines.
- Author
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Januchowski R, Zawierucha P, Ruciński M, and Zabel M
- Subjects
- Antineoplastic Agents pharmacology, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Humans, Ovarian Neoplasms drug therapy, Drug Resistance, Neoplasm, Extracellular Matrix Proteins genetics, Gene Expression Profiling methods, Oligonucleotide Array Sequence Analysis methods, Ovarian Neoplasms genetics
- Abstract
Ovarian cancer is the most lethal gynecological malignancy. Multiple drug resistance (MDR) development leads to resistance of cancer cells to chemotherapy. Microarray methods can provide information regarding new candidate genes that can play a role in resistance to cytostatic drugs. Extracellular matrix (ECM) can influence drug resistance by inhibiting the penetration of the drug into cancer tissue as well as increased apoptosis resistance. In the present study, we report changes in the ECM and related gene expression pattern in methotrexate-, cisplatin-, doxorubicin-, vincristine-, topotecan- and paclitaxel-resistant variants of the W1 ovarian cancer cell line. The resistant variants of the W1 cell line were generated by stepwise selection of cells with an increasing concentration of the indicated drugs. Affymetrix GeneChip® Human Genome U219 Array Strips were used for hybridizations. Independent t-tests were used to determinate the statistical significance of results. Genes whose expression levels were higher than the assumed threshold (upregulated, >5-fold and downregulated, <5-fold) were visualized using the scatter plot method, selected and listed in the tables. Among the investigated genes, expression of 24 genes increased, expression of 14 genes decreased and expression of three genes increased or decreased depending on the cell line. Among the increased genes, expression of 10 increased very significantly, >20-fold. These genes were: ITGB1BP3, COL3A1, COL5A2, COL15A1, TGFBI, DCN, LUM, MATN2, POSTN and EGFL6. The expression of seven genes decreased very significantly: ITGA1, COL1A2, LAMA2, GPC3, KRT23, VIT and HMCN1. The expression pattern of ECM and related genes provided the preliminary view into the role of ECM components in cytostatic drug resistance of cancer cells. The exact role of the investigated genes in drug resistance requires further investigation.
- Published
- 2014
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23. Drug transporter expression profiling in chemoresistant variants of the A2780 ovarian cancer cell line.
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Januchowski R, Zawierucha P, Ruciński M, Andrzejewska M, Wojtowicz K, Nowicki M, and Zabel M
- Subjects
- Biological Transport, Cell Line, Tumor, Down-Regulation, Drug Resistance, Neoplasm, Female, Gene Expression Regulation, Neoplastic, Humans, Oligonucleotide Array Sequence Analysis, Ovarian Neoplasms drug therapy, Up-Regulation, ATP-Binding Cassette Transporters genetics, Antineoplastic Agents pharmacology, Membrane Transport Proteins genetics, Ovarian Neoplasms genetics
- Abstract
Ovarian cancer is characterized by the higher mortality among gynecological cancers. In results of MDR development during chemotherapy cancer cells become resistant to further treatment. Microarray techniques can provide information about MDR development at gene expression level. ABC and SLC transporters are most important proteins responsible for this phenomenon. In this study changes of ABC and SLC genes expression pattern in drugs resistant sublines of the A2780 ovarian cancer cell line were demonstrated. The cytostatic resistant sublines were generated by culture of A2780 cell line with an increasing concentration of the indicated drugs. As screening methods, we used Affymetrix U219 Human Genome microarrays. Independent t-tests were used to determinate statistical significances of results. Genes that expression levels were higher than assumed threshold (upregulated above threefold and downregulated under -3 fold) were visualized using scatter plot method, selected and listed in table. Between the ABC genes increased expression of seven genes and decreased expression of three genes were observed. Expression of two genes was increased or decreased depending on the cell line. We observed significant (more than tenfold) increase in expression of four ABC genes: ABCA8, ABCB1, ABCB4 and ABCG2 and decreased expression of ABCA3 gene. We also observed changes in expression of 32 SLC genes. Between them we observe increased expression of 17 genes and decreased expression of 15 genes. Expression of four genes was increased or decreased dependent on cell line. The expression of nine SLC genes increased or decreased very significantly (more than tenfold). Five genes were significantly upregulated: SLC2A9, SLC16A3, SLC16A14, SLC38A4 and SLC39A8. Four additional genes were significantly downregulated: SLC2A14, SLC6A15, SLC8A1 and SLC27A2. Expression profiles of these genes give strong arguments for assumption of correlation between expression of ABC and SLC genes and drug resistance phenomenon. Identifying correlations between specific drug transporters and cytostatic drug resistance will require further investigation., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)
- Published
- 2014
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24. Extracellular matrix proteins expression profiling in chemoresistant variants of the A2780 ovarian cancer cell line.
- Author
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Januchowski R, Zawierucha P, Ruciński M, Nowicki M, and Zabel M
- Subjects
- Cell Line, Tumor, Extracellular Matrix Proteins genetics, Female, Gene Expression Profiling, Humans, Neoplasm Proteins genetics, Oligonucleotide Array Sequence Analysis, Ovarian Neoplasms genetics, Ovarian Neoplasms pathology, Drug Resistance, Neoplasm, Extracellular Matrix Proteins biosynthesis, Gene Expression Regulation, Neoplastic, Neoplasm Proteins biosynthesis, Ovarian Neoplasms metabolism
- Abstract
Ovarian cancer is the leading cause of death among gynaecological malignancies. Extracellular matrix (ECM) can affect drug resistance by preventing the penetration of the drug into cancer cells and increased resistance to apoptosis. This study demonstrates alterations in the expression levels of ECM components and related genes in cisplatin-, doxorubicin-, topotecan-, and paclitaxel-resistant variants of the A2780 ovarian cancer cell line. Affymetrix Gene Chip Human Genome Array Strips were used for hybridisations. The genes that had altered expression levels in drug-resistant sublines were selected and filtered by scatter plots. The genes that were up- or downregulated more than fivefold were selected and listed. Among the investigated genes, 28 genes were upregulated, 10 genes were downregulated, and two genes were down- or upregulated depending on the cell line. Between upregulated genes 12 were upregulated very significantly--over 20-fold. These genes included COL1A2, COL12A1, COL21A1, LOX, TGFBI, LAMB1, EFEMP1, GPC3, SDC2, MGP, MMP3, and TIMP3. Four genes were very significantly downregulated: COL11A1, LAMA2, GPC6, and LUM. The expression profiles of investigated genes provide a preliminary insight into the relationship between drug resistance and the expression of ECM components. Identifying correlations between investigated genes and drug resistance will require further analysis.
- Published
- 2014
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25. Microarray-based detection and expression analysis of ABC and SLC transporters in drug-resistant ovarian cancer cell lines.
- Author
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Januchowski R, Zawierucha P, Andrzejewska M, Ruciński M, and Zabel M
- Subjects
- ATP-Binding Cassette Transporters genetics, Animals, Cell Line, Tumor, Female, GABA Plasma Membrane Transport Proteins biosynthesis, Humans, Ovarian Neoplasms genetics, ATP-Binding Cassette Transporters biosynthesis, Drug Resistance, Neoplasm genetics, GABA Plasma Membrane Transport Proteins genetics, Gene Expression Regulation, Neoplastic, Ovarian Neoplasms metabolism, Protein Array Analysis methods
- Abstract
Multiple drug resistance of cancer cells is multifactorial. A microarray technique may provide information about new candidate genes playing a role in drug resistance. Drug membrane transporters from ABC and SLC families play a main role in this phenomenon. This study demonstrates alterations in ABC and SLC gene expression levels in methotrexate, cisplatin, doxorubicin, vincristine, topotecan and paclitaxel-resistant variant of W1 ovarian cancer cell line. Resistant W1 cell lines were derived by stepwise selection of cells in increasing concentration of drugs. Affymetrix GeneChip(®) Human Genome U219 Array Strip was used for hybridizations. Statistical significance was determined by independent sample t-test. The genes having altered expression levels in drug-resistant sublines were selected and filtered by scater plot. Genes up/downregulated more than threefolds were selected and listed. Among ABC genes, seven were upregulated and three were downregulated. Three genes: ABCB1, ABCB4 and ABCG2 were upregulated very significantly (over tenfold). One ABCA8 was significantly downregulated. Among 38 SLC genes, 18 were upregulated, 16 were downregulated and four were up- or downregulated dependent on the cell line. Expression of 10 SLC genes was changed very significantly (over tenfold). Four genes were significantly increased: SLC6A1, SLC9A2, SLC12A1, SLC16A6 and six genes were significantly decreased: SLC2A14, SLC7A3, SLC7A8, SLC7A11, SLC16A14, SLC38A9. Based on the expression profiles, our results provide a preliminary insight into the relationship between drug resistance and expression of membrane transporters involved in drug resistance. Correlation of specific drug transporter with drug resistance requires further analysis., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)
- Published
- 2013
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26. Genetically modified human myoblasts with eNOS may improve regenerative ability of myogenic stem cells to infarcted heart.
- Author
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Janeczek A, Zimna A, Rozwadowska N, Fraczek M, Kucharzewska P, Ruciński M, Mietkiewski T, Kolanowski T, Malcher A, and Kurpisz M
- Subjects
- Animals, Apoptosis genetics, Cell Cycle genetics, Cell Proliferation, Cells, Cultured, Electroporation, Endothelial Cells cytology, Humans, Mice, Myoblasts, Skeletal cytology, Myoblasts, Skeletal metabolism, Myoblasts, Smooth Muscle cytology, Myoblasts, Smooth Muscle metabolism, Neovascularization, Physiologic genetics, Oxidative Stress genetics, Regeneration genetics, Stem Cell Transplantation, Transfection, Umbilical Veins cytology, Vascular Endothelial Growth Factor A, Genetic Therapy, Myoblasts, Skeletal transplantation, Myoblasts, Smooth Muscle transplantation, Myocardial Infarction therapy, Nitric Oxide Synthase Type III genetics, Stem Cells cytology
- Abstract
Background: Modern therapies of post infarcted heart failure are focused on perfusion improvement of the injured myocardium. This effect can be achieved by, among other means, implanting stem cells which could be genetically modified with factors inducing the formation of new blood vessels in the post infarction scar area. Combined stem cell and gene therapy seems to be a promising strategy to heal an impaired myocardium. The creation of new blood vessels can be indirectly stimulated via factors inducing vascular endothelial growth factor synthesis, for example endothelial nitric oxide synthase (eNOS). The product of this enzyme, nitric oxide, is a molecule that can influence numerous physiological activities; it can contribute to vasodilation, stimulation of endothelial cell growth, prevention of platelet aggregation and leukocyte adhesion to the endothelium., Aim: To verify the pro-angiogenic and regenerative potential of human primary myoblasts and murine myoblast cell line C2C12 transiently transfected with eNOS gene., Methods: Stem cells (either human or murine) were maintained in standard in vitro conditions. Next, both types of myoblasts were modified using electroporation and lipofection (human and murine cells), respectively. The efficacy of the transfection method was evaluated using flow cytometry. The concentration of eNOS protein was measured by ELISA immunoassay. The biological properties of modified cells were assessed using an MTT proliferation test and DAPI cell cycle analysis. To verify the influence of oxidative stress on myoblasts, cytometric tests using Annexin V and propidium iodide were applied. To check possible alterations in myogenic gene expression of stem cells transduced by genetic modification, the myogenic regulatory factors were evaluated by real-time PCR. The function of genetic modification was confirmed by a HUVEC capillary sprouting test using myoblasts supernatants., Results: Electroporation turned out to be an efficient transfection method. High amounts of secreted protein were obtained (in the range 2,000 pg/mL) in both cell types studied. Moreover, the functionality of gene overexpression product was confirmed in capillary development assay. Human myoblasts did not exhibit any changes in cell cycle; however, eNOS transfected murine myoblasts revealed a statistically significant reduction in cell cycle ratio compared to controls (p < 0.001). In the case of myogenic gene expression, a decrease in Myogenin level was only detected in the human transfected myoblast population (p < 0.05)., Conclusions: The results of our study may suggest that transplantation of myoblasts overexpressing eNOS could be promising for cell therapy in regenerating the post infarction heart.
- Published
- 2013
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27. Expression of ghrelin receptor (GHSR-1a) in rat epididymal spermatozoa and the effects of its activation.
- Author
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Łukaszyk A, Kotwicka M, Jankowska A, Kasprzak A, Ruciński M, Sterzyńska K, Ziółkowska A, Sawiński P, and Ruchala M
- Subjects
- Acrosome, Animals, Biological Transport, Caspase 3 metabolism, Epididymis cytology, Ghrelin pharmacology, Male, Phosphatidylserines genetics, Phosphatidylserines metabolism, Rats, Receptors, Ghrelin genetics, Seminiferous Epithelium metabolism, Sperm Motility physiology, Epididymis physiology, Gene Expression Regulation physiology, Receptors, Ghrelin metabolism, Spermatozoa physiology
- Abstract
In this study we demonstrated the expression of the ghrelin receptor GHSR-1a in rat spermatids and epididymal spermatozoa, as well as some effects of ghrelin on the spermatozoa in vitro. For the demonstration of GHSR-1a the immunocytochemical, immunofluorescence and Western blotting techniques were applied using three different types of antibodies. The response of spermatozoa to ghrelin was tested in a series of in vitro experiments and their effects were evaluated using confocal microscopy and flow cytometry. GHSR-1a protein was found as expressed in the Golgi and acrosomes of spermatids and acrosome regions or the head cell membrane of epididymal spermatozoa. The GHSR-1a expression in spermatozoa was also confirmed by Western blot. No differences were found in percentage of spermatozoa showing annexin-V binding and expression of active form caspase-3 between control and ghrelin-treated spermatozoa. This result may indicate no pro-apoptotic effects of ghrelin neither at 10(-9) nor 10(-6)mol/L concentration. Ghrelin (10(-6)mol/L) increased free intracellular calcium ion concentration in the rat spermatozoa. Moreover, stimulation with 10(-6)mol/L ghrelin increased, while 10(-4)mol/L ghrelin decreased the number of spermatozoa showing progressive motility. In conclusion, the expression of the GHSR-1a receptor in spermatozoa, as well as ghrelin influences on sperm motility and intracellular calcium ion concentration suggest that such biological effects of ghrelin may be produced under in vivo conditions., (Copyright © 2012 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.)
- Published
- 2012
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28. Neuropeptide B and W regulate leptin and resistin secretion, and stimulate lipolysis in isolated rat adipocytes.
- Author
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Skrzypski M, Pruszyńska-Oszmałek E, Ruciński M, Szczepankiewicz D, Sassek M, Wojciechowicz T, Kaczmarek P, Kołodziejski PA, Strowski MZ, Malendowicz LK, and Nowak KW
- Subjects
- Adipocytes drug effects, Animals, Male, Neuropeptides pharmacology, Rats, Rats, Wistar, Adipocytes metabolism, Leptin metabolism, Lipolysis drug effects, Neuropeptides metabolism, Resistin metabolism
- Abstract
Neuropeptide B (NPB) and W (NPW) regulate food intake and energy homeostasis in humans via two G-protein-coupled receptor subtypes, termed as GPR7 and GPR8. Rodents express GPR7 only. In animals, NPW decreases insulin and leptin levels, whereas the deletion of either NPB or GPR7 leads to obesity and hyperphagia. Metabolic and endocrine in vitro activities of NPW/NPB in adipocytes are unknown. We therefore characterize the effects of NPB and NPW on the secretion and expression of leptin and resistin, and on lipolysis, using rat adipocytes. Isolated rat adipocytes express GPR7 mRNA. NPB and NPW are expressed in macrophages and preadipocytes but are absent in mature adipocytes. Both, NPB and NPW reduce the secretion and expression of leptin from isolated rat adipocytes. NPB stimulates the secretion and expression of resistin, whereas both, NPB and NPW increase lipolysis. Our study demonstrates for the first time that NPB and NPW regulate the expression and secretion of leptin and resistin, and increase lipolysis in isolated rat adipocytes. These effects are presumably mediated via GPR7. The increase of resistin secretion, stimulation of lipolysis and the decrease of leptin secretion may represent mechanisms, through which NPB and NPW can affect glucose and lipid homeostasis, and food intake in rodents., (Copyright © 2012 Elsevier B.V. All rights reserved.)
- Published
- 2012
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29. Immunohistochemical and hybridocytochemical study on ghrelin signalling in the rat seminiferous epithelium.
- Author
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Łukaszyk A, Rafińska L, Sawiński P, Kasprzak A, Olejniczak K, Ruciński M, Ruchała M, and Sowiński J
- Subjects
- Animals, Ghrelin biosynthesis, Immunohistochemistry, Male, Rats, Rats, Wistar, Receptors, Ghrelin biosynthesis, Receptors, Ghrelin metabolism, Spermatogenesis, Ghrelin metabolism, Seminiferous Epithelium metabolism, Signal Transduction
- Abstract
The results of presented study demonstrate expression of ghrelin, its functional receptor GHSR-1a and their genes in spermatogenic cells of rat testis suggesting their functioning within seminiferous epithelium. The immunohistochemical and hybrydocytochemical expression, of proteins and transcripts, was estimated taking into account the cycle of seminiferous epithelium and phases of spermatogenesis. Both transcripts and ghrelin was found to show nuclear expression and scarcely cytoplasmic. Expression of genes for ghrelin and GHSR-1a was shown in early spermatocytes and round spermatids representing transcriptional phases of meiosis and spermiogenesis. Ghrelin was evidenced to show nuclear expression in two stage-specific windows, in late spermatogonia, in spermatocytes up to early pachytenes, and again in spermatids of acrosome and early maturation phase of spermiogenesis. In late pachytenes, secondary spermatocytes, round spermatids, maturing spermatids and spermatozoa the reaction is lacking. With two types of antibodies against the GHSR-1a used the two different patterns of immunostaining was evidenced suggesting two isoforms of GHSR-1a. The first evidenced GHSR-1a in cytoplasm of spermatocytes, cell membrane and acrosomes of spermatids, Sertoli cell processes and heads of spermatozoa. With second type of antibodies the immunostaining marks all steps of evolution of acrosome in spermatids. It is believed that site of ghrelin expression in seminiferous epithelium may indicate its role in local regulations, not excepting the intracellular signalling. Immunostaining pattern for GHSR-1a seems to suggest both its participation in the cross-talk among the cells and also process of furnishing gametes with GHSR-1a for its response to ghrelin in seminal plasma or female reproductive tract.
- Published
- 2009
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30. Expression of the beacon gene in the rat pancreatic islets: opposite effects of beacon (47-73) protein (ubiquitin-like protein 5) on insulin secretion in vivo and insulin release by isolated islets.
- Author
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Nowak KW, Ruciński M, Kaczmarek P, Szkudelski T, and Malendowicz LK
- Subjects
- Animals, Blood Glucose analysis, Cells, Cultured drug effects, Cells, Cultured metabolism, Depression, Chemical, Dose-Response Relationship, Drug, Female, Gene Expression Regulation, Glucose pharmacology, In Vitro Techniques, Injections, Subcutaneous, Insulin Secretion, Islets of Langerhans drug effects, Nerve Tissue Proteins genetics, Nerve Tissue Proteins pharmacology, Nerve Tissue Proteins physiology, Peptide Fragments pharmacology, RNA, Messenger biosynthesis, Rats, Rats, Wistar, Reverse Transcriptase Polymerase Chain Reaction, Ubiquitins pharmacology, Insulin metabolism, Islets of Langerhans metabolism, Nerve Tissue Proteins biosynthesis
- Abstract
Objective: Beacon gene expression is elevated in the hypothalamus of the Israeli sand rat (Psammomys obesus), an animal that is used as a polygenic animal model of obesity and NIDDM. We performed studies aimed at investigating the expression of beacon mRNA and protein in pancreatic islets of the rat and the possible beacon protein effects on insulin secretion., Methods: Rat pancreatic islets were isolated by the collagenase digestion technique. Beacon mRNA expression was demonstrated in isolated islets using RT-PCR and beacon-like immunoreactivity using immunocytochemistry (ICC) on a sections of Bouin-fixed pancreas. Isolated islets were incubated with 1-100 nmol/L beacon (47-73) protein in normoglycemic medium. Adult female rats were subcutaneously injected with beacon (47-73) at doses 0.35 or 0.7 nmol/100 g body weight and killed after 30 and 60 minutes., Results: RT-PCR results indicate the presence of beacon mRNA in isolated rat pancreatic islets. Beacon-like immunoreactivity is present in all cell types of the Langerhans islet. Beacon inhibits insulin secretion from isolated islets. In contrast, a bolus administration of beacon at a lower dose notably stimulates blood insulin concentration at 30 and 60 minutes of the experiment while the higher dose does not change insulinemia. None of the treatment had an effect on blood glucose concentration., Conclusion: This study demonstrated the presence of beacon mRNA in isolated rat islets as well as a direct inhibitory effect of beacon protein on insulin secretion by isolated rat pancreatic islets. The data obtained suggest that beacon may be involved in physiologic regulation of insulin secretion.
- Published
- 2004
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31. [Perioperative sedation and analgesia for complications from arthroscopy and arthrotomy of the knee joint].
- Author
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Kozierowski T, Wołowicka L, Małysz A, Ruciński M, and Rzymski S
- Subjects
- Administration, Oral, Alprazolam administration & dosage, Bupivacaine administration & dosage, Epinephrine administration & dosage, Female, Fentanyl administration & dosage, Humans, Lidocaine administration & dosage, Male, Midazolam administration & dosage, Pain Measurement, Pain, Postoperative etiology, Patient Satisfaction, Preanesthetic Medication, Premedication, Analgesia methods, Arthroscopy adverse effects, Conscious Sedation methods, Knee Joint surgery, Pain, Postoperative prevention & control
- Abstract
Estimation of the quality of the epidural anaesthesia of the patients sedated with Alprazolam and Midazolam in premedication before arthroscopy or arthrotomy of the knee was the goal of our study. Forty six (34 men and 12 women) ASA physical status 1-2 patients were divided into groups depending on the drugs orally applied in premedication (Alprazolam 0.5 mg, n = 29 or Midazolam 15 mg, n = 17) and of the kind of analgesia. The patients subjected to arthroscopy were treated with a single-shot epidural analgesia (n = 38), while those subjected to arthrotomy--with a continuous epidural analgesia (n = 8). 2% Lignocain with addition of Epinephrine and Fentanyl was used in the perioperative analgesia, while 0.25% Bupivacain with addition of Morphine was used in the postoperative period when continuous epidural analgesia was applied. The ISAS, VAS and Ramsey scales were used and the data were analysed with the Kormogolov test. The perioperative sedation in arthroscopy and arthrotomy of the knee is good without any significant differences associated with a kind of the drugs applied. The single-shot epidural anaesthesia is inadequate only during a prolonged arthroscopy of the knee. The postoperative continuous epidural analgesia, expressed in the VAS scale, was inadequate. A level of general satisfaction of the patients of the sedation and analgesia, expressed in the points of the ISAS scale, was satisfactorily good.
- Published
- 2001
32. Comparative morphology of the spinal ganglia in the different segments of the spinal cord in sheep.
- Author
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Hereć S, Ruciński M, and Milart Z
- Subjects
- Animals, Ganglia, Spinal anatomy & histology, Sheep anatomy & histology, Spinal Cord anatomy & histology
- Published
- 1996
33. [Occurrence of risk factors for atherosclerosis in young women with myocardial infarction].
- Author
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Smielak-Korombel W, Franaszek J, Kanas P, Kulon I, Pytel T, and Ruciński M
- Subjects
- Adult, Female, Humans, Risk Factors, Arteriosclerosis etiology, Myocardial Infarction
- Abstract
Occurrence of atherosclerosis risk factors was analyzed in 16 women aged 26-45 with myocardial infarction. The control group consisted of women of the same age and profession. Stated risk factors were in order: hypertension, diabetes mellitus, positive family history, fat metabolism disturbances, stress and cigarette smoking. The most hazardous association was fat metabolism disturbances connected with hypertension, positive family history or hypertension or excess sensibility to stress were also frequently observed. Authors stated three, four, five and even six risk factors together in 64.3% of women after myocardial infarction.
- Published
- 1989
34. [Effect of glucagon on bile lipids].
- Author
-
Dura K, Wedrychowicz A, and Ruciński M
- Subjects
- Adult, Aged, Female, Humans, Male, Middle Aged, Stimulation, Chemical, Bile analysis, Glucagon pharmacology, Lipids analysis
- Published
- 1976
35. [Therapy of diabetes mellitus with Silubin-retard].
- Author
-
Huczek-Glebocki J, Piotrowska R, and Ruciński M
- Subjects
- Adult, Aged, Blood Glucose analysis, Drug Synergism, Female, Glucose Tolerance Test, Humans, Insulin therapeutic use, Male, Middle Aged, Biguanides therapeutic use, Diabetes Mellitus drug therapy
- Published
- 1970
Catalog
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