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Background: Colorectal cancer (CRC) presents significant treatment challenges due to its high heterogeneity and complex intercellular interactions. Further exploration of CRC subtypes and interactions among tumor-specific clusters will facilitate the development of personalized treatment strategies. Methods: Single-cell RNA sequencing and bulk RNA sequencing datasets were integrated to determine CRC metabolic subtypes by hierarchical clustering. The analysis was further extended to cellchat, pseudotime, immune infiltration, and clinicopathological relevance to explore the characteristics of secreted frizzled related protein 2 (SFRP2) + cancer-associated fibroblast (CAF) clusters, and validated by spatial transcriptomics (ST), in vivo experiments, and multiple immunohistochemistry (mIHC). Results: CRC samples were stably classified into three heterogeneous metabolic subtypes, each exhibiting different microenvironment and CAF heterogeneity, particularly in the distribution of SFRP2 + CAF, which was aligned with metabolic activity. SFRP2 + CAF exhibits high extracellular matrix (ECM) activity and is closely involved in cellular communication, not only promoting the malignant progression of cancer cells but also inducing the differentiation of Tregs. Compared to responders of chemotherapy, the proportion of SFRP2 + CAFs is significantly increased in non-responders. Importantly, mIHC and ST analyses confirm that cancer cells with low expression of agmatinase (AGMAT) can recruit SFRP2 + CAFs, and Treg infiltration surrounding SFRP2 + CAFs was observed. AGMAT combined with oxaliplatin showed the best efficacy in vivo, which may be associated with the inhibition of SFRP2 + CAF infiltration. Conclusions: Our study identified and described the potential protumor biological properties of SFRP2 + CAFs, and AGMAT may be a valuable target for disrupting their properties. [ABSTRACT FROM AUTHOR]

The epithelial and mesenchymal features of colorectal adenocarcinoma (CRAC) cell lines were compared in two-dimensional (2D) and three-dimensional (3D) cultures. In 2D cultures, the three CRAC cell lines exhibited epithelial characteristics with high E-cadherin and low vimentin levels, whereas two exhibited mesenchymal traits with opposite expression patterns. In 3D cultures using low-attachment plates, mesenchymal cells from 2D cultures showed reduced vimentin mRNA levels. Morphologically, the five CRAC cell lines appeared similarly shaped in 2D culture but formed different structures in 3D culture. Epithelial DLD-1 and mesenchymal COLO-320 cells produced large granular spheres, whereas epithelial HCT-15 cells formed small solid spheres. Tubular structures were observed in epithelial CACO-2 and mesenchymal SW480 spheres. Desmosome-like structures developed in epithelial CRAC cells, whereas entosis was observed in CACO-2, HCT-15, and SW480 cells. The Ki-67-positive proliferating cell count varied in 2D and 3D cultures of epithelial cells but remained high and unchanged in mesenchymal cells. These findings suggest that while CRAC cells display distinct epithelial and mesenchymal properties in 2D cultures, they form diverse 3D structures, irrespective of these traits. [ABSTRACT FROM AUTHOR]

Dysregulated pH is now recognised as a hallmark of cancer. Recent evidence has revealed that the endosomal pH regulator Na+/H+ exchanger NHE9 is upregulated in colorectal cancer to impose a pseudo-starvation state associated with invasion, highlighting an underexplored mechanistic link between adaptive endosomal reprogramming and malignant transformation. In this study, we use a model that quantitatively captures the dynamics of the core regulatory network governing epithelial mesenchymal plasticity. The model recapitulated NHE9-induced calcium signalling and the emergence of migratory phenotypes in colorectal cancer cells. Model predictions were compared with patient data and experimental results from RNA sequencing analysis of colorectal cancer cells with stable NHE9 expression. Mathematical analyses identified that tumours leverage elevated NHE9 levels to delay the transition of cells to a mesenchymal state and allow for metastatic progression. Ectopic expression of NHE9 is sufficient to induce loss of epithelial nature but does not fully couple with gain of mesenchymal state, resulting in a hybrid epithelial-mesenchymal population with increased aggressiveness and metastatic competence. Higher NHE9 expression is associated with cancer cell migration, and the effect appears to be independent of hypoxia status. Our data suggests that alterations in endosomal pH, an evolutionarily conserved starvation response, may be hijacked by colorectal cancer cells to drive phenotypic plasticity and invasion. We propose that cancer cells rewire their endosomal pH not only to meet the demands of rapid cell proliferation, but also to enable invasion, metastasis, and cell survival. Endosomal pH may be an attractive therapeutic target for halting tumour progression. [ABSTRACT FROM AUTHOR]

Recent research has shown that Doublecortin-like kinase 1 (DCLK1) is overexpressed in different types of cancer. It has recently been described as a cancer stem cells (CSCs) marker, is associated with carcinogenesis, and positively correlates with infiltration of multiple immune cell types in some cancers. However, studies focused on assessing DCLK1 expression in HCC are limited, and the role of DCLK1 in HCC tumor immunity remains to be determined. In this study, we used a modified model of the resistant hepatocyte (MRHM) to evaluate DCLK1 expression in HCC. Furthermore, DCLK1 expression in HCC was analyzed using TIMER 2.0, UALCAN, GEPIA, GEO, and HPA web-based tools. Correlations between DCLK1 expression and clinicopathological factors in patients were analyzed using the UALCAN web-based tool. Finally, correlations between DCLK1 and immune infiltrates were investigated using the TIMER 2.0 and TISIDB web-based tools. The results showed that DCLK1 is significantly overexpressed during progression of the HCC carcinogenic process in the MRHM. DCLK1 is overexpressed in HCC according to multiple publics web-based tools, and its overexpression is associated with cancer stage. Furthermore, DCLK1 expression was correlated with infiltration levels of multiple immune cells, immunomodulatory factors, immunoinhibitors, MHC molecules, chemokines, receptors, and immune cell-specific markers. These results suggest that DCLK1 is a potential prognostic biomarker that determines cancer progression and correlates with immune cell infiltration in HCC. [ABSTRACT FROM AUTHOR]

Objective: Colorectal cancer (CRC) is a type of cancer with high mortality and widespread metastasis. The aim was to determine the expression levels of apoptosis-related genes in CRC cells treated with Allium polyanthum Schult plant extract and the combination of this plant extract with docetaxel. Methods: Expression analyses of caspase-2 (CASP2), nuclear factor NF-kappa-B1 (NFKB1), apoptosis regulator BAX and proto-oncogene MYC genes that play a role in apoptosis in the CRC cell line HT-29 and the healthy colon cell line CCD-18Co, which were treated with only Allium polyanthum Schult plant extract and docetaxel together with this plant extract, were performed using the real-time polymerase chain reaction (RT-PCR) method. Bioinformatics analysis of relevant genes was performed using various databases. Results: CASP2, MYC, NFKB1 and BAX gene expression was significantly decreased in CRC cells treated with the combination of Allium polyanthum Schult extract and docetaxel compared to healthy cells. Accordingly, only the extract of Allium polyanthum Schult plant significantly reduced the expression of apoptosis-related genes in HT-29 cells compared to the extract combined with docetaxel. As a result of bioinformatics analysis, it was found that CASP2, MYC, NFKB1 and BAX proteins interact with each other and the expression levels of their genes are associated with survival. In addition, the methylation status of CASP2 and NFKB1 has the potential to change protein levels by affecting epigenetic mechanisms in CRC. Conclusion: According to the information in the literature, it has been reported that Allium species affect genes in apoptotic pathways. Accordingly, Allium polyanthum Schult alone and in combination with docetaxel should be supported by further studies. [ABSTRACT FROM AUTHOR]

The aim of the study was to assess the survival rates and surgery-related toxicity in patients with locally advanced squamous cell vulvar cancer (LAVC) managed by upfront chemoradiation (CRT) with/without following by surgery. CRT is the primary treatment for patients with unresectable locally advanced squamous cell vulvar carcinoma (LAVC), followed by surgery in case of residual tumor. Patients with AJCC stage II-IV squamous cell vulvar carcinoma referred to Gynecologic Oncology Unit at Fondazione Policlinico Universitario Agostino Gemelli I.R.C.C.S. from January 2016 to February 2023, managed by upfront CRT, were included. 63 patients were included, 21 (33 %) had complete response (cCR) to CRT, 26 (41 %) had partial response (cPR), 1 (2 %) stable disease (cSD), 15 (24 %) had disease progression (cPD). In the whole population, cPR/SD and cPD were associated with reduced PFS (p < 0.001) and overall survival (OS) (p < 0.001), p16 expression was associated with improved PFS (p < 0.001) and OS (p = 0.001). Among patients with clinical residual disease after CRT, 23 patients undergoing surgery experienced improved PFS (p = 0.003) and OS (p = 0.003) compared to those receiving other treatments. Eight (35 %) patients experienced severe (grade ≥ III) postoperative complications; vulvar and groin wound dehiscence/infection were the most common complications; one (4 %) patient died in the postoperative. Patients with pathological residual disease experienced worse PFS (p = 0.013) and OS (p = 0.034). Clinical response to CRT and p16 expression strongly predict survival in LAVC. Surgery for residual disease might be associated with improved survival but is burdened by high rates of complications. Pathologic residual disease correlates with high recurrence rates and poor survival. • Response to chemoradiation and p16 expression predict survival in locally advanced squamous cell vulvar cancer. • Among patients with residual disease, radical surgery is associated with improved survival. • Radical surgery after chemoradiation is associated with high rates of postoperative complications. • Pathologic residual disease is associated with high rates of recurrence and death from disease. [ABSTRACT FROM AUTHOR]

Molecular analysis of breast cancer cells by niosomal encapsulated Withania somnifera (WS) extract is the focus of this investigation. Characterization of ASH-loaded niosomes produced by a thin film technique showed that they have an appropriate zeta potential, small particle size, and low polydispersity index. The concentration-dependent suppression of cell proliferation was seen in MCF-7 breast cancer cells treated with ASH-NIO, with an IC50 of 24.13 µM, and olaparib at values of 10.45 µM. Molecular investigation showed that ASH-NIO treated cells had lower levels of B-cell lymphoma 2 (Bcl2) mRNA expression compared to untreated cells, and upregulated levels of Tumor protein 53 (P53), Bcl-2-associated X protein (Bax), caspase-9, and caspase-3. These results provide more evidence that ASH-NIO has potential anti-cancer effects and may be useful as an adjunctive treatment for breast cancer. [ABSTRACT FROM AUTHOR]

This study aims to investigate the role and prognostic significance of long non-coding RNAs (lncRNAs) associated with disulfidptosis in colon adenocarcinoma (COAD). The TCGA database's clinical data and transcriptome profiles were employed. Analysis of previous studies identified 10 disulfidptosis-related genes (DRGs). We used these genes to construct a signature that could independently and accurately predict the prognosis of patients with COAD. The Kaplan-Meier (K-M) curve analysis showed that the lower-risk group had a better prognosis. With the help of multivariate Cox regression analysis, the risk score produced from the patient's signature might independently predict the outcomes. Utilizing a nomogram, the receiver operating characteristic (ROC) curve, and principal component analysis (PCA), the signature's predictive ability was also confirmed. It's interesting to note that immunotherapy, especially PD-1 immune checkpoint suppression, was more likely to benefit low-risk patients. The IC50 levels for certain anticancer agents were lower in the high-risk group. Finally, qRT-PCR analyses in colon cancer cell lines revealed elevated levels of lncRNAs CASC9, ZEB1-AS1, ATP2A1-AS1, SNHG7, AL683813.1, and AP003555.1, and reduced levels of FAM160A1-DT and AC112220.2, compared to normal cell lines. This signature offers insights into prognosis, tumor microenvironment, and options for immunotherapy and antitumor drugs in patients with COAD. [ABSTRACT FROM AUTHOR]

The brown alga Padina boryana Thivy has been studied for its bioactive properties. This study aimed to evaluate the antioxidant, anticancer, alpha-glucosidase inhibitory, anti-inflammatory, antimicrobial, and hepatoprotective activities of Padina boryana extracts. The antioxidant activity was investigated using DPPH and ABTS radical scavenging, and ferric reducing power assays. The anticancer activity was determined via cytotoxic activity on human lung carcinoma (A549) and human embryonic kidney (HEK 293) cancer cell lines. Alpha-glucosidase inhibitory activity was determined using standard method, anti-inflammatory activity was assessed via nitric oxide (NO) inhibition assay, antimicrobial activity was assessed using the broth microdilution assay, and the hepatoprotective activity was evaluated using CCl4-induced hepatotoxicity in HepG2 cells. Results showed that the algal extracts have low DPPH scavenging activity, but effective ABTS scavenging activity with EC50 value of 52.09 µg/mL for the ethyl acetate (EA) extract. The EA extract had the highest ferric-reducing power with EC50 of 61.7 µg/mL. The petroleum ether (PE) extract had the highest cytotoxic effect with EC50 values of 38.19 µg/mL and 33.28 µg/mL against A549, and HEK 293 cells, respectively. All extracts showed significant alpha-glucosidase inhibitory activity, with EA being the most potent (IC50 = 3.01 µg/mL). The PE extract showed notable NO inhibitory activity with IC50 of 50.91 µg/mL. No significant antimicrobial activity was observed except for EtOH and PE extracts, which inhibited Staphylococcus aureus and Bacillus subtilis. The extracts did not demonstrate significant hepatoprotective effect. These findings highlight the potential of Padina boryana as a valuable source of natural therapeutic agents. [ABSTRACT FROM AUTHOR]

The detection and analysis of cancer cell exosomes with high sensitivity and precision are pivotal for the early diagnosis and treatment strategies of prostate cancer. To this end, a microfluidic chip, equipped with a cactus-like array substrate (CAS) based on surface-enhanced Raman spectroscopy (SERS) was designed and fabricated for the detection of exosome concentrations in Lymph Node Carcinoma of the Prostate (LNCaP). Double layers of polystyrene (PS) microspheres were self-assembled onto a polyethylene terephthalate (PET) film to form an ordered cactus-like nanoarray for detection and analysis. By combining EpCAM aptamer-labeled SERS nanoprobes and a CD63 aptamer-labeled CAS, a 'sandwich' structure was formed and applied to the microfluidic chips, further enhancing the Raman scattering signal of Raman reporter molecules. The results indicate that the integrated microfluidic sensor exhibits a good linear response within the detection concentration range of 105 particles μL−1 to 1 particle μL−1. The detection limit of exosomes in cancer cells can reach 1 particle μL−1. Therefore, we believed that the CAS integrated microfluidic sensor offers a superior solution for the early diagnosis and therapeutic intervention of prostate cancer. [ABSTRACT FROM AUTHOR]

Abstract Circulating tumor cell separation has been the focus of numerous studies owing to its importance in the diagnosis, prognosis, and therapy of cancer. This study reports a highly efficient microfluidic device that integrates a specialized dielectrophoresis configuration, namely the facing-electrode configuration dielectrophoresis (FEC-DEP) structure, to isolate circulating tumor cells (CTCs) from various blood components, including red blood cells, white blood cells, and platelets. The FEC-DEP design features a bottom-slanted electrode array positioned parallel to a basic rectangular top electrode. A non-homogeneous electric field is produced between these parallel electrodes, generating dielectrophoretic forces acting on cells. Consequently, when the FEC-DEP is integrated into a flow, it can direct various biological objects in the flow along separate trajectories. As a result, cells with comparable characteristics might move together within a similar path. This configuration may simplify the microfabrication process and lessen dependency on particle position within the microchannel. The separation process was numerically analyzed using the finite element method, and device parameters were optimized to obtain high-efficiency and high-purity cell separation. The simulations show that the microfluidic device may effectively enrich tumor cells in a label-free and non-invasive manner, with a high-efficiency rate of almost 80%.

Serum and glucocorticoid-regulated kinase 1 (SGK1) plays a multifaceted role in cancer progression and treatment resistance. Its importance stems from its role in several cellular functions essential to cancer development, including metabolism, apoptosis, cell survival, and proliferation. In cancer, SGK1 expression is often dysregulated, leading to its overactivation or overexpression in many malignancies. This dysregulation can promote tumor growth, invasion, and metastasis through several mechanisms. We perform a bibliometric analysis to explore the role of SGK1 in cancer in the last ten years. All publications related to this topic were retrieved from the Scopus database. Microsoft Office Excel 2021 and VOSviewer (version 1.6.20) were used for the bibliometric analysis. The filtered search identified 5322 articles published between 2013 and 2023. Regarding research and scientific collaboration in this field, China is in the lead, followed by the USA, Germany, the UK, Japan, and Italy. In summary, SGK1 plays a critical role in cancer by regulating cell survival, proliferation, metabolism, and stemness. Its dysregulation promotes tumor growth, metastasis, and resistance to therapy. Targeting SGK1 is promising for enhancing cancer treatment efficacy, but further research is needed to fully understand its mechanisms and develop effective therapeutic strategies. Our study offers significant contributions to the field of cancer research by providing a comprehensive understanding of SGK1's role in cancer biology and highlighting its potential as a therapeutic target. These insights can inform future studies and guide the development of novel cancer prevention, diagnosis, and treatment approaches. [ABSTRACT FROM AUTHOR]

Neuroendocrine tumor (NET) is a rare malignant tumor, notably small cell lung cancer (SCLC), a type of lung neuroendocrine tumor, which has a survival rate of less than 7%. Although various biomarkers including CHGA (Chromogranin A), INSM1 (Insulinoma-associated protein 1), and SYP (Synaptophysin) are extensively used for the diagnostic testing of NET, their diverse specificities and sensitivities are acknowledged as limitations. Here, we demonstrate that RepID (Replication initiation determinant protein), a component of CRL4 (Cullin-RING ubiquitin E3 ligase 4), holds promise as a biomarker for identifying NET and SCLC. Analysis of the Cancer Cell Line Encyclopedia (CCLE) via the CellMinerCDB portal reveals a high correlation between RepID transcript levels and mRNA expression of NE signature genes. Additionally, RepID protein is highly expressed in SCLC patient tissues and a subset of SCLC cell lines. Viability analysis following treatment with pevonedistat and SZL-P1-41 in SCLC cell lines and human SCLC-organoid models indicates that RepID expression determines the sensitivity to CRL-targeting anti-cancer drugs. These findings suggest that RepID represents a novel biomarker for NET and SCLC, and insights from RepID research in these cancers could lead to innovative therapeutic strategies. [ABSTRACT FROM AUTHOR]

Metabolic dysregulation is a key driver of cellular senescence, contributing to the progression of systemic aging. The heterogeneity of senescent cells and their metabolic shifts are complex and unexplored. A microfluidic SlipChip integrated with surface‐enhanced Raman spectroscopy (SERS), termed SlipChip‐SERS, is developed for single‐cell metabolism analysis. This SlipChip‐SERS enables compartmentalization of single cells, parallel delivery of saponin and nanoparticles to release intracellular metabolites and to realize SERS detection with simple slipping operations. Analysis of different cancer cell lines using SlipChip‐SERS demonstrated its capability for sensitive and multiplexed metabolic profiling of individual cells. When applied to human primary fibroblasts of different ages, it identified 12 differential metabolites, with spermine validated as a potent inducer of cellular senescence. Prolonged exposure to spermine can induce a classic senescence phenotype, such as increased senescence‐associated β‐glactosidase activity, elevated expression of senescence‐related genes and reduced LMNB1 levels. Additionally, the senescence‐inducing capacity of spermine in HUVECs and WRL‐68 cells is confirmed, and exogenous spermine treatment increased the accumulation and release of H2O2. Overall, a novel SlipChip‐SERS system is developed for single‐cell metabolic analysis, revealing spermine as a potential inducer of senescence across multiple cell types, which may offer new strategies for addressing ageing and ageing‐related diseases. [ABSTRACT FROM AUTHOR]

The tumor microenvironment (TME) encompasses the complex and diverse surroundings in which tumors arise. Emerging insights highlight the TME's critical role in tumor development, progression, metastasis, and treatment response. Consequently, the TME has attracted significant research and clinical interest, leading to the identification of numerous novel therapeutic targets. Advances in molecular technologies now enable detailed genomic and transcriptional analysis of cancer cells and the TME and the integration of microenvironmental data to the tumor genomic landscape. This comprehensive review discusses current progress in targeting the TME for drug development, addressing associated challenges, strategies for modulating the pro-tumor microenvironment, and the discovery of new targets. [ABSTRACT FROM AUTHOR]

Lipid extraction of complex biological samples is essential for high‐quality data in liquid chromatography‐mass spectrometry (LC‐MS)‐based lipidomics. This study introduces a three‐phase liquid extraction (3PLE)‐ultra‐high‐performance LC coupled with quadrupole time‐of‐flight tandem MS method. This method was successfully applied to lipidomics analysis of breast cancer cells, including highly metastatic MDA‐MB‐231 and slightly metastatic MCF7 cells. The 3PLE method employed an n‐hexane/methyl tert‐butyl ether/acetonitrile/water solvent system that formed one aqueous and two organic phases. Neutral and polar lipids were enriched in the upper and middle organic phases, respectively, and combined for detection, thereby reducing analysis time. Compared with the Bligh and Dyer method, 3PLE achieved higher sensitivity and detected more features, with over a 50% increase in the relative abundance of nearly 50% of the differential lipids. In total, 21 differential lipids were identified in the MDA‐MB‐231 group and 22 in the MCF7 group compared to normal breast epithelial cells (MCF10A). Pathway analysis suggested that lipid changes in breast cancer cells were associated with glycerophospholipid metabolism, arachidonic acid metabolism, sphingolipid metabolism, and linoleic acid metabolism. The study presents a highly efficient lipidomics method, providing a scientific foundation for understanding breast cancer pathogenesis and aiding in diagnosis. [ABSTRACT FROM AUTHOR]

Introduction: Fetal microchimerism could be involved in the regulation of breast cancer oncogenesis. CD34+ cells could be of a particular interest as up to 12% of the CD34+ population in maternal blood are of fetal origin. The aim of this research was to analyze the impact of umbilical cord blood (UCB) CD34+ on MCF-7 and MDA-MB-231 breast cancer cell lines, in order to uncover novel biological mechanisms and suggest novel treatment options for breast cancer. Methods: UCB CD34+ cells were obtained from healthy women at full-term delivery. Direct cultures were grown with MCF-7 and MDA-MB-231 cells. Proliferation, migration, invasion, and transcriptomic analysis of breast cancer cell lines were compared between cultures exposed and nonexposed to UCB CD34+ cells. Interactions between UCB CD34+ and breast cancer cells were analyzed under fluorescent microscopy. Functional analyses were generated with QIAGEN’s Ingenuity Pathway Analysis (IPA) and Gene Set Enrichment Analysis (GSEA). Results: Direct contact between UCB CD34+ and breast cancer cell lines induced a reduction in the proliferative capacities of MCF-7 and MDA-MB-231 and diminished the migration abilities of MDA-MB-231 cells. In 3D coculture, UCB CD34+ cells were attracted by tumor spheroids and incorporated into tumor cells. These cell-to-cell interactions were responsible for transcriptome modifications coherent with observed functional modifications. Among the cytokines secreted by UCB CD34+, IFN-γ was identified as a potential upstream regulator responsible for the molecular modifications observed in transcriptomic analysis of MCF-7 breast cancer cells exposed to UCB CD34+ cells, as was IL-17A in MDA-MB-231 cells. Conclusion: Direct cell-to-cell contact induced functional modifications in breast cancer cells. Interactions between UCB CD34+ and breast cancer cells could induce cell fusion and signal transmission via cytokines. Further analysis of direct cell-to-cell interactions should be performed at a molecular level to further understand the potential role of fetal CD34+ cells in breast cancer. [ABSTRACT FROM AUTHOR]

Introduction: Real-world data on programmed death-ligand 1 (PD-L1) testing and biomarker-guided treatment in patients with advanced/metastatic non-small cell lung cancer (mNSCLC) are lacking. Methods: This non-interventional, retrospective study used patient data from the lung cancer registry of the Sotiria Hospital in Greece. Included patients had mNSCLC and initiated first line (L) treatment from August 2016 (when pembrolizumab gained reimbursement) to August 2019; patient follow-up ended in August 2020. The study mainly assessed the PD-L1 testing rates and the treatment(s) prescribed by epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) status, and PD-L1 tumor proportion score (TPS). Data were descriptively analyzed. Kaplan-Meier estimates of timeto- event outcomes were generated. Results: Of 705 patients, 304 (43.1%) were tested for PD-L1 expression during observation. PD-L1 testing rates reached 73.8% across quarters Q1– Q3 of 2019; most patients were tested at 1L. At 1L, 59.3% of patients with EGFR/ALK negative or unknown status and PD-L1 TPS ≥50% received PD-1/ PD-L1 inhibitors; 90.6% of these patients received pembrolizumab. Regardless of PD-L1 expression levels, most patients with EGFR/ALK negative or unknown status received PD-1/PD-L1 inhibitors at 2L. The numerically highest 1L median overall survival for patients with EGFR/ALK negative or unknown status was attained with PD-1/PD-L1 inhibitors, with overall survival (OS) of 26.7 months. Conclusions: PD-L1 testing rates increased rapidly following the reimbursement of pembrolizumab in the 1L treatment of patients with mNSCLC. Patients with EGFR/ALK negative or unknown status and PD-L1 TPS ≥50% most frequently received PD-1/PD-L1 inhibitors at 1L, and attained longer OS compared with other therapies. [ABSTRACT FROM AUTHOR]

Pregnancy‐associated breast cancer has been increasing. In this study, we analyzed patients with breast cancer that occurred during pregnancy (PrBC) and compared their genetic profiles with those of patients with breast cancer that did not occur during pregnancy, within 1 year after childbirth nor during lactation (non‐PrBC). We performed gene expression analyses of patients with PrBC and non‐PrBC using microarrays and qRT‐PCR. Microarray analysis showed that 355 genes were upregulated in the luminal‐type PrBC group compared to those in the non‐PrBC group. The C‐X‐C motif chemokine ligand 13 (CXCL13) gene was the most upregulated in the PrBC group compared to that in the non‐PrBC group, especially in the luminal A‐type (p = 0.016). This result was corroborated by the qRT‐PCR analysis of microdissected cancer cells (p < 0.001). A negative correlation was observed between CXCL13 and estrogen receptor 1 (ESR1) mRNA expression levels in luminal A‐type breast carcinoma (p < 0.001). Our results provide clues for a better understanding of breast cancer pathogenesis during pregnancy. [ABSTRACT FROM AUTHOR]

Aims: Nanoparticles due to their wide applications in medicine,industry,and biotechnology, have attracted many scientists' attentions. Recently, nanoparticles especially selenium nanoparticles are widely used to diagnosis and cancer treatment. The aim of this study was to evaluate the cytotoxic and anticancer effects of selenium nanoparticles on colon cancer cell line and analysis of CAD (Caspase Activated DNase) gene expression. Material and methods: In this study, colon cancer HT29 and normal HEK293 cell lines were purchased from the Pasteur Institute Cell Bank of Tehran and treated with selenium nanoparticles overnight. The cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) (Gibco, Scotland) medium with 10% FBS serum and 1% streptomycin antibiotic (Gibco, Scotland). The cells were then stored at 37°C. In this study, cytotoxic effect of Selenium NPs was evaluated on HT29 and HEK293 cells using MTT (3-(4, 5-Dimethyltetrazollium Bromide) assay. Subsequently, they were treated with selenium nanoparticles in different concentrations (0, 7.81, 15.62, 31.25, 62.5, 125, 250 and 500 µg/mL) for 24 hours. To solubilize the viable cells formazan crystals production, we added 100 μl/well of dimethyl sulfoxide (DMSO) to them. After treatment of HT29 cells with IC50 concentration, the total RNA was extracted and cDNA synthesized. Moreover, CAD gene expression was evaluated using Real Time PCR method. The data was evaluated by ABI StepOne utilizing the Applied Biosystems qRT-PCR (ABI 7300 system, Applied Biosystems). The quantification of the mode of Selenium NPs-induced cell death in the HT29 cells were ascertained using flow cytometry followed by staining with fluorescein isothiocyanate (FITC)-Annexin V and propidium iodide (PI) staining. Finally, the study of apoptosis and necrosis of Selenium NPs was evaluated using flow cytometry method. Data analysis was statistically determined by using One-way analysis of variance (ANOVA) with SPSS/22 software followed by a Tukey test. [ABSTRACT FROM AUTHOR]

Context: Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is the central system in epigenomic exploration. Chromatin immunoprecipitation coupled with sequencing (ChIP-seq) is an important technology to identify the genome-wide location of DNA-binding proteins such as histones proteins, transcription factors, RNA polymerase, or any protein of interest. ChIP-seq has been used to study the binding sites and efficacy of drugs in cancer cell lines etc. Aims: In current research, breast cancer cell line data have been used to study the effect PADI2 (peptidyl arginine deiminase) gene in the progression of breast cancer. Further, this ChIP-seq data have also been used to study the binding site of Amanitin drug in breast cancer. Settings and Design: Breast cancer ChIP-seq data have been retrieved from the European Nucleotide Archive database with project Id PRJNA415426 short read archive. Four samples of FASTQ files were used and analyzed for the genome-wide analysis. Materials and Methods: Galaxy server (https://usegalaxy.org/) was used for complete ChIP-seq data analysis; different tools such as fast-quality control (QC), multi-QC, Bowtie2, model-based analysis of ChIP-sequencing, and ChIPseeker tools were used for motif enrichment and functional analysis. Motif analysis was done through the Multiple Expectation maximizations for Motif Elicitation database (https://meme-suite.org/meme/db/motifs). Results: Computational investigation demonstrates the binding sequences of the T47-D breast cancer cell line as TTTTGTATTTTTAGT, and this motif occurs 2123 times in the Homo Sapiens reference genome that is hg19. Conclusions: This research classifies the binding site and affinity of the T47-D human breast cancer cell line. Further, wet laboratory studies are required to verify the function of the predicted motifs and their importance in drug development or research in breast cancer. [ABSTRACT FROM AUTHOR]

We reported previously that in preclinical models, BMP4 is a potent inhibitor of breast cancer metastasis and that high BMP4 protein levels predict favourable patient outcomes. Here, we analysed a breast cancer xenograft with or without enforced expression of BMP4 to gain insight into the mechanisms by which BMP4 suppresses metastasis. Transcriptomic analysis of cancer cells recovered from primary tumours and phosphoproteomic analyses of cancer cells exposed to recombinant BMP4 revealed that BMP4 inhibits cholesterol biosynthesis, with many genes in this biosynthetic pathway being downregulated by BMP4. The treatment of mice bearing low-BMP4 xenografts with a cholesterol-lowering statin partially mimicked the anti-metastatic activity of BMP4. Analysis of a cohort of primary breast cancers revealed a reduced relapse rate for patients on statin therapy if their tumours exhibited low BMP4 levels. These findings indicate that BMP4 may represent a predictive biomarker for the benefit of additional statin therapy in breast cancer patients. [ABSTRACT FROM AUTHOR]

This paper introduces an innovative optical biosensor designed for multimode refractive index (MMRI) detection, employing split ring resonator (SRR) technology for enhanced sensitivity in cancer cell analysis. The proposed biosensor comprises two concentric SRRs, one resembling a cube and the other a cylinder, each incorporating an air gap. Rigorous analysis and optimization of dimensional parameters were conducted using the finite difference time domain (FDTD) numerical solution method. The simulation results demonstrated that the biosensor achieves exceptional sensitivity and an optimal figure of merit (FOM). Specifically, the first mode of the biosensor exhibited a sensitivity exceeding 80 nm/RIU and an FOM surpassing 20 RIU⁻1, while the second mode demonstrated a sensitivity of 570 nm/RIU and an FOM exceeding 4 RIU⁻1. These results indicate that the first mode benefits from a higher FOM, enhancing precision and accuracy, while the second mode benefits from high sensitivity, enabling the detection of minute refractive index changes. With a primary focus on early cancer cell detection and timely treatment initiation to address the global cancer mortality burden, the biosensor's high-quality factor (Q-factor) and FOM promise significant advancements in cancer diagnostics and therapeutic interventions. [ABSTRACT FROM AUTHOR]

Neuroendocrine tumors (NETs) of the pancreas are rare neoplasms that present complex challenges to diagnosis and treatment due to their indolent course. The incidence of pancreatic neuroendocrine tumors has increased significantly over the past two decades. A limited number of pancreatic neuroendocrine cell lines are currently available for the research. Here, we present 3D-iNET ORION, a novel 3-dimensional (spheroid) cell line, isolated from human pancreatic neuroendocrine tumor liver metastasis. Three-dimensionally grown (3D) cancer cell lines have gained interest over the past years as 3D cancer cell lines better recapitulate the in vivo structure of tumors, and are more suitable for in vitro and in vivo experiments. 3D-iNET ORION cancer cell line showed high potential to form tumorspheres when embedded in Matrigel matrix and expresses synaptophysin and EpCAM. Electron microscopy analysis of cancer cell line proved the presence of dense neurosecretory granules. When xenografted into athymic mice, 3D-iNET ORION cells produce slow-growing tumors, positive for chromogranin and synaptophysin. Human Core Exome Panel Analysis has shown that 3DiNET ORION cell line retains the genetic aberration profile detected in the original tumor. In conclusion, our newly developed neuroendocrine cancer cell line can be considered as a new research tool for in vitro and in vivo experiments. [ABSTRACT FROM AUTHOR]

Lung cancer stands as the primary cause of cancer-related death across the globe. The standard therapeutic approach for lung cancer involves concurrent chemoradiotherapy, with consideration of prophylactic cranial irradiation for younger or well-performing patients. In this study, we aimed to investigate prognostic factors and the impacts of different treatment methods on overall survival for stage IIIA small cell lung cancer in Taiwan. We obtained data from the Taiwan Cancer Registry, which included clinical and pathology data of 579 stage IIIA small cell lung cancer patients from January 2010 to December 2018, for this retrospective study. The enrolled patients had data on age, sex, Charlson Comorbidity Index score, histologic grading, clinical T, clinical N, clinical stage, treatment modality, and overall survival time. We compared overall survival among different subgroups to assess the impacts of these prognostic factors. The five-year survival rate for all patients was 20.57%, with a median survival time of 15.79 months. The data suggest that Charlson Comorbidity Index score, histologic grade, and clinical stage subgroups did not reach statistically significant differences. During the multivariate analysis, age over 70 years, sex, and treatment method were determined to be statistically significant independent prognostic factors. Patients who underwent surgical intervention exhibited significantly better outcomes compared to those who did not undergo operation.. In conclusion, stage IIIA small cell lung cancer is a highly heterogeneous disease. Operation should be considered as one of the alternative treatments in stage IIIA Small cell lung cancer patients. [ABSTRACT FROM AUTHOR]

Background: Deletions and partial losses of chromosome 7 (chr7) are frequent in acute myeloid leukemia (AML) and are linked to dismal outcome. However, the genomic landscape and prognostic impact of concomitant genetic aberrations remain incompletely understood. Methods: To discover genetic lesions in adult AML patients with aberrations of chromosome 7 [abn(7)], 60 paired diagnostic/remission samples were investigated by whole-exome sequencing in the exploration cohort. Subsequently, a gene panel including 66 genes and a SNP backbone for copy-number variation detection was designed and applied to the remaining samples of the validation cohort. In total, 519 patients were investigated, of which 415 received intensive induction treatment, typically containing a combination of cytarabine and anthracyclines. Results: In the exploration cohort, the most frequently mutated gene was TP53 (33%), followed by epigenetic regulators (DNMT3A, KMT2C, IDH2) and signaling genes (NRAS, PTPN11). Thirty percent of 519 patients harbored ≥ 1 mutation in genes located in commonly deleted regions of chr7—most frequently affecting KMT2C (16%) and EZH2 (10%). KMT2C mutations were often subclonal and enriched in patients with del(7q), de novo or core-binding factor AML (45%). Cancer cell fraction analysis and reconstruction of mutation acquisition identified TP53 mutations as mainly disease-initiating events, while del(7q) or −7 appeared as subclonal events in one-third of cases. Multivariable analysis identified five genetic lesions with significant prognostic impact in intensively treated AML patients with abn(7). Mutations in TP53 and PTPN11 (11%) showed the strongest association with worse overall survival (OS, TP53: hazard ratio [HR], 2.53 [95% CI 1.66–3.86]; P < 0.001; PTPN11: HR, 2.24 [95% CI 1.56–3.22]; P < 0.001) and relapse-free survival (RFS, TP53: HR, 2.3 [95% CI 1.25–4.26]; P = 0.008; PTPN11: HR, 2.32 [95% CI 1.33–4.04]; P = 0.003). By contrast, IDH2-mutated patients (9%) displayed prolonged OS (HR, 0.51 [95% CI 0.30–0.88]; P = 0.0015) and durable responses (RFS: HR, 0.5 [95% CI 0.26–0.96]; P = 0.036). Conclusion: This work unraveled formerly underestimated genetic lesions and provides a comprehensive overview of the spectrum of recurrent gene mutations and their clinical relevance in AML with abn(7). KMT2C mutations are among the most frequent gene mutations in this heterogeneous AML subgroup and warrant further functional investigation. [ABSTRACT FROM AUTHOR]

The aetiology of bone metastasis in prostate cancer (PCa) remains unclear. This study aims to identify hub genes involved in this process. We utilized machine learning, GO, KEGG, GSEA, Single‐cell analysis, ROC methods to identify hub genes for bone metastasis in PCa using the TCGA and GEO databases. Potential drugs targeting these genes were identified. We validated these results using 16 specimens from patients with PCa and analysed the relationship between the hub genes and clinical features. The impact of APOC1 on PCa was assessed through in vitro experiments. Seven hub genes with AUC values of 0.727–0.926 were identified. APOC1, CFH, NUSAP1 and LGALS1 were highly expressed in bone metastasis tissues, while NR4A2, ADRB2 and ZNF331 exhibited an opposite trend. Immunohistochemistry further confirmed these results. The oxidative phosphorylation pathway was significantly enriched by the identified genes. Aflatoxin B1, benzo(a)pyrene, cyclosporine were identified as potential drugs. APOC1 expression was correlated with clinical features of PCa metastasis. Silencing APOC1 significantly inhibited PCa cell proliferation, clonality, and migration in vitro. This study identified 7 hub genes that potentially facilitate bone metastasis in PCa through mitochondrial metabolic reprogramming. APOC1 emerged as a promising therapeutic target and prognostic marker for PCa with bone metastasis. [ABSTRACT FROM AUTHOR]

Analysis of single cancer cells is critical to obtain accurate patient diagnosis and prognosis. In this work, we report the selective dielectrophoretic capture and electrochemical analysis of single melanoma cells at an array of interlocked spiral bipolar electrodes (iBPEs). Following dielectrophoretic capture, individual melanoma cells are hydrodynamically transferred into picoliter‐scale chambers for subsequent analysis. The interlocked spiral end of the iBPE (the sensing pole) is utilized to read out an electrochemical enzyme‐linked immunosorbent assay (eELISA), which quantifies the expression of a cell surface antigen, melanoma cell adhesion marker (MCAM). The opposite pole of each BPE is located in a fluidically isolated compartment containing reagents for electrogenerated chemiluminescence (ECL), such that luminescence reports iBPE current. In a preliminary device design, the ECL intensity was insufficient to detect MCAM expression on single cells. To achieve single‐cell analysis, we decreased the gap size between the interlocked spirals tenfold (5.0 μm to 0.5 μm), thereby creating a more sensitive biosensor by enhanced redox cycling of the product of eELISA. This work is significant because it allows for the selective isolation and sensitive analysis of individual melanoma cells in a device amenable to point‐of‐care (POC) application by combining dielectrophoresis (DEP) with interdigitated bipolar electrodes (IDBPEs). [ABSTRACT FROM AUTHOR]

Human papillomaviruses (HPVs) cause most cervical cancers and an increasing number of anogenital and oral carcinomas, with most cases caused by HPV16 or HPV18. HPV hijacks host signalling pathways to promote carcinogenesis. Understanding these interactions could permit identification of much-needed therapeutics for HPV-driven malignancies. The Hippo signalling pathway is important in HPV+ cancers, with the downstream effector YAP playing a pro-oncogenic role. In contrast, the significance of its paralogue TAZ remains largely uncharacterised in these cancers. We demonstrate that TAZ is dysregulated in a HPV-type dependent manner by a distinct mechanism to that of YAP and controls proliferation via alternative cellular targets. Analysis of cervical cancer cell lines and patient biopsies revealed that TAZ expression was only significantly increased in HPV18+ and HPV18-like cells and TAZ knockdown reduced proliferation, migration and invasion only in HPV18+ cells. RNA-sequencing of HPV18+ cervical cells revealed that YAP and TAZ have distinct targets, suggesting they promote carcinogenesis by different mechanisms. Thus, in HPV18+ cancers, YAP and TAZ play non-redundant roles. This analysis identified TOGARAM2 as a previously uncharacterised TAZ target and demonstrates its role as a key effector of TAZ-mediated proliferation, migration and invasion in HPV18+ cancers. The Hippo pathway transcription factors YAP and TAZ are often thought to play redundant roles in cancer progression. Here, Patterson et al demonstrate that TAZ is specifically required in HPV18+ cervical cancer, a subtype associated with worse prognosis. [ABSTRACT FROM AUTHOR]

Breast cancer (BC) cells characteristics have played a crucial role in clinical strategies decisionmaking and patient outcome prediction. Although there have been several studies examining this, there are still relatively few that analyze stem cell subpopulations or protein biomarkers. The conducted research sought to elucidate breast cancer subtype characteristics based on immunohistochemical analysis and cancer stem cell presence based on flow cytometry analysis in three breast cancer samples from Bandung, Indonesia. A flow cytometry analysis was conducted to determine how much CD44/CD24 was expressed as a marker of cancer stem cell in isolated BC cells using various cell references, such as cell lines of SKBR-3, MDA-MB-468, and Hs587T. Immunohistochemistry analysis was carried out to investigate the expression of p53, HER2, PGR, Ki67, and ER in sample tissue sections. According to histological analysis, 2 samples were solid mammary carcinoma, and 1 sample was mammary fibroadenoma. Immunohistochemistry and histological analysis showed that all BC tissues are classified as Luminal B. In addition, a large percentage of CD44+/CD24- subpopulation was found in isolated BC cells (> 90 % in patients 1 and 3; > 60 % in patient 2). All samples showed similarity to Hs587T cell line characteristic. This study has successfully characterized solid mammary tumors and mammary fibroadenoma with luminal B characteristics. All specimens contained a high proportion of cancer stem cells. [ABSTRACT FROM AUTHOR]

Natural killer cell‐derived extracellular vesicles (NK‐EVs) are candidate biotherapeutics against various cancers. However, standardised potency assays are necessary for a reliable assessment of NK‐EVs' cytotoxicity. This study aims to thoroughly evaluate a highly sensitive resazurin phenoxazine‐based cell viability potency assay (measurement of the cellular redox metabolism) for quantifying the cytotoxicity of NK‐EVs against leukaemia K562 cells (suspension model) and breast cancer MDA‐MB‐231 cells (adherent model) in vitro. The assay was evaluated based on common analytical parameters setforth by regulatory guidelines, including specificity, selectivity,accuracy, precision, linearity, range and stability. Our results revealed that this resazurin‐based cell viability potency assay reliably and reproducibly measured a dose‐response of NK‐EVs' cytotoxic activity against both cancer models. The assay showed precision with 5% and 20% variation for intra‐run and inter‐run variability. The assay signal showed specificity and selectivity of NK‐EVs against cancer target cells, as evidenced by the diminished viability of cancer cells following a 5‐hour treatment with NK‐EVs, without any detectable interference or background. The linearity analysis of target cancer cells revealed strong linearity for densities of 5000 K562 and 1000 MDA‐MB‐231 cells per test with a consistent range. Importantly, NK‐EVs' dose‐response for cytotoxicity showed a strong correlation (|ρ| ∼ 0.8) with the levels of known cytotoxic factors associated with the NK‐EVs' corona (FasL, GNLY, GzmB, PFN and IFN‐γ), thereby validating the accuracy of the assay. The assay also distinguished cytotoxicity changes in degraded NK‐EVs, indicating the ability of the assay to detect the potential loss of sample integrity. Compared to other commonly reported bioassays (i.e., flow cytometry, cell counting, lactate dehydrogenase release assay, DNA‐binding reporter assay and confluence assay), our results support this highly sensitive resazurin‐based viability potency assay as a high‐throughput and quantitative method for assessing NK‐EVs' cytotoxicity against both suspension and adherent cancer models for evaluating NK‐EVs' biotherapeutics. [ABSTRACT FROM AUTHOR]

Prostate cancer is a prevalent carcinoma among males, and conventional treatment options are often limited. Cytotoxic chemotherapy, despite its drawbacks, remains a mainstay. We propose a targeted co‐delivery approach using nanoscale delivery units for Oncolytic measles virus (OMV) and vincristine (VC) to enhance treatment efficacy. The HA‐coated OMV + VC‐loaded TCs nanoformulation is designed for targeted oncolytic activity in prostate cancer. The CD44 expression analysis in prostate cancer cell lines indicates a significantly high expression in PC3 cells. The optimization of nanoformulations using Design of Expert (DOE) is performed, and the preparation and characterization of HA‐coated OMV + VC‐loaded TCs nanoformulations are detailed showing average particle size 397.2 ± 0.01 nm and polydispersity index 0.122 with zeta potential 19.7 + 0.01 mV. Results demonstrate successful encapsulation efficiency with 2.4 × 106 TCID50/Ml and sustained release of OMV and VC from the nanoformulation for up to 72 h. In vitro, assays reveal potent anticancer activity at 10 ± 0.71% cell viability in PC3 cells compared to 73 ± 0.66% in HPrEC and significant morphological changes at 90 µg/ml in dose and time‐dependent manner. The co‐formulation showed positive cell death 49.5 ± 0.02% at 50 µg PI/ml in PBS and 54.3% cell cycle arrest at the G2/M phase, 8.1% G0/G1 and 5.7% at S phase, with significant mitochondrial membrane potential (MMP) at 50 µg/ml, as assessed by flow cytometry (FACS). The surface‐integrating ligand approach enhances the targeted delivery of the oncolytic virus and chemotherapeutic drug, presenting a potential alternative for prostate cancer treatment and suggested that co‐administering VC and OMV in a nanoformulation could improve therapeutic outcomes while reducing chemotherapeutic drug doses. [ABSTRACT FROM AUTHOR]

Background: Acute myeloid leukemia (AML) is a hematopoietic malignancy with poor outcomes, especially in older AML patients. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered a promising anticancer drug because it selectively induces the extrinsic apoptosis of tumor cells without affecting normal cells. However, clinical trials have shown that the responses of patients to TRAIL are significantly heterogeneous. It is necessary to explore predictable biomarkers for the preselection of AML patients with better responsiveness to TRAIL. Here, we investigated the critical role of tumor protein p53 inducible nuclear protein 2 (TP53INP2) in the AML cell response to TRAIL treatment. Methods: First, the relationship between TP53INP2 and the sensitivity of AML cells to TRAIL was determined by bioinformatics analysis of Cancer Cell Line Encyclopedia datasets, Cell Counting Kit-8 assays, flow cytometry (FCM) and cell line-derived xenograft (CDX) mouse models. Second, the mechanisms by which TP53INP2 participates in the response to TRAIL were analyzed by Western blot, ubiquitination, coimmunoprecipitation and immunofluorescence assays. Finally, the effect of TRAIL alone or in combination with the BCL-2 inhibitor venetoclax (VEN) on cell survival was explored using colony formation and FCM assays, and the effect on leukemogenesis was further investigated in a patient-derived xenograft (PDX) mouse model. Results: AML cells with high TP53INP2 expression were more sensitive to TRAIL in vitro and in vivo. Gain- and loss-of-function studies demonstrated that TP53INP2 significantly enhanced TRAIL-induced apoptosis, especially in AML cells with nucleophosmin 1 (NPM1) mutations. Mechanistically, cytoplasmic TP53INP2 maintained by mutant NPM1 functions as a scaffold bridging the ubiquitin ligase TRAF6 to caspase-8 (CASP 8), thereby promoting the ubiquitination and activation of the CASP 8 pathway. More importantly, simultaneously stimulating extrinsic and intrinsic apoptosis signaling pathways with TRAIL and VEN showed strong synergistic antileukemic activity in AML cells with high levels of TP53INP2. Conclusion: Our findings revealed that TP53INP2 is a predictor of responsiveness to TRAIL treatment and supported a potentially individualized therapeutic strategy for TP53INP2-positive AML patients. [ABSTRACT FROM AUTHOR]

This article, titled "Role of PSMD5 Protein-Protein Interactions in Proteasome Assembly and Cancer," explores the relationship between proteotoxic stress and the levels of PSMD5, a chaperone involved in proteasome assembly. The study found that many cancer cells have decreased levels of PSMD5, potentially as a response to proteotoxic stress. Overexpression of PSMD5 in breast cancer cells was shown to decrease proteasome levels and activity. The authors suggest that inhibiting PSMD5 levels and interactions could help regulate homeostasis and overcome proteotoxicity. Further research using affinity purification coupled mass-spectrometry (AP-MS) is needed to identify other interacting partners of PSMD5 and their role in proteostasis regulation. [Extracted from the article]

We aimed to determine whether monitoring tumor-derived exosomal microRNAs (miRNAs) could be used to assess radiotherapeutic sensitivity in patients with locally advanced esophageal squamous cell carcinoma (ESCC). RNA sequencing was employed to conduct a comparative analysis of miRNA expression levels during radiotherapy, focusing on identifying miRNAs associated with progression. Electron microscopy confirmed the existence of exosomes, and co-cultivation assays and immunofluorescence validated their capacity to infiltrate macrophages. To determine the mechanism by which exosomal miR-143-3p regulates the interplay between ESCC cells and M2 macrophages, ESCC cell-derived exosomes were co-cultured with macrophages. Serum miR-143-3p and miR-223-3p were elevated during radiotherapy, suggesting resistance to radiation and an unfavorable prognosis for ESCC. Increased levels of both miRNAs independently predicted shorter progression-free survival (p = 0.015). We developed a diagnostic model for ESCC using serum microRNAs, resulting in an area under the curve of 0.751. Radiotherapy enhanced the release of miR-143-3p from ESCC cell-derived exosomes. Immune cell infiltration analysis at the Cancer Genome Atlas (TCGA) database revealed that ESCC cell-derived miR-143-3p triggered M2 macrophage polarization. Mechanistically, miR-143-3p upregulation affected chemokine activity and cytokine signaling pathways. Furthermore, ESCC cell exosomal miR-143-3p could be transferred to macrophages, thereby promoting their polarization. Serum miR-143-3p and miR-223-3p could represent diagnostic and prognostic markers for patients with ESCC undergoing radiotherapy. Unfavorable prognosis could be linked to the increased levels of ESCC cell-derived exosomal miR-143-3p, which might promote tumor progression by interacting with macrophages. [ABSTRACT FROM AUTHOR]

The molecular system regulating cellular mechanical properties remains unexplored at single-cell resolution mainly due to a limited ability to combine mechanophenotyping with unbiased transcriptional screening. Here, we describe an electroporation-based lipid-bilayer assay for cell surface tension and transcriptomics (ELASTomics), a method in which oligonucleotide-labelled macromolecules are imported into cells via nanopore electroporation to assess the mechanical state of the cell surface and are enumerated by sequencing. ELASTomics can be readily integrated with existing single-cell sequencing approaches and enables the joint study of cell surface mechanics and underlying transcriptional regulation at an unprecedented resolution. We validate ELASTomics via analysis of cancer cell lines from various malignancies and show that the method can accurately identify cell types and assess cell surface tension. ELASTomics enables exploration of the relationships between cell surface tension, surface proteins, and transcripts along cell lineages differentiating from the haematopoietic progenitor cells of mice. We study the surface mechanics of cellular senescence and demonstrate that RRAD regulates cell surface tension in senescent TIG-1 cells. ELASTomics provides a unique opportunity to profile the mechanical and molecular phenotypes of single cells and can dissect the interplay among these in a range of biological contexts. The molecular system regulating cell surface mechanics remains largely unexplored at single-cell resolution. Here, the authors report a high-throughput single-cell assay, ELASTomics, which integrates mechanical phenotyping with unbiased transcriptomics. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) shows promise in tumor treatment, particularly when combined with nanotechnology. This study examines the impact of deep learning, particularly the Cellpose algorithm, on the comprehension of cancer cell responses to PDT. The Cellpose algorithm enables robust morphological analysis of cancer cells, while logistic growth modelling predicts cellular behavior post-PDT. Rigorous model validation ensures the accuracy of the findings. Cellpose demonstrates significant morphological changes after PDT, affecting cellular proliferation and survival. The reliability of the findings is confirmed by model validation. This deep learning tool enhances our understanding of cancer cell dynamics after PDT. Advanced analytical techniques, such as morphological analysis and growth modeling, provide insights into the effects of PDT on hepatocellular carcinoma (HCC) cells, which could potentially improve cancer treatment efficacy. In summary, the research examines the role of deep learning in optimizing PDT parameters to personalize oncology treatment and improve efficacy. [ABSTRACT FROM AUTHOR]

Discovering effective anti-cancer agents poses a formidable challenge given the limited efficacy of current therapeutic modalities against various cancer types due to intrinsic resistance mechanisms. Cancer immunochemotherapy is an alternative strategy for breast cancer treatment and overcoming cancer resistance. Human Indoleamine 2,3-dioxygenase (hIDO1) and human Tryptophan 2,3-dioxygenase 2 (hTDO2) play pivotal roles in tryptophan metabolism, leading to the generation of kynurenine and other bioactive metabolites. This process facilitates the de novo synthesis of Nicotinamide Dinucleotide (NAD), promoting cancer resistance. This study identified a new dual hIDO1/hTDO2 inhibitor using a drug repurposing strategy of FDA-approved drugs. Herein, we delineate the development of a ligand-based pharmacophore model based on a training set of 12 compounds with reported hIDO1/hTDO2 inhibitory activity. We conducted a pharmacophore search followed by high-throughput virtual screening of 2568 FDA-approved drugs against both enzymes, resulting in ten hits, four of them with high potential of dual inhibitory activity. For further in silico and in vitro biological investigation, the anti-hypercholesterolemic drug Pitavastatin deemed the drug of choice in this study. Molecular dynamics (MD) simulations demonstrated that Pitavastatin forms stable complexes with both hIDO1 and hTDO2 receptors, providing a structural basis for its potential therapeutic efficacy. At nanomolar (nM) concentration, it exhibited remarkable in vitro enzyme inhibitory activity against both examined enzymes. Additionally, Pitavastatin demonstrated potent cytotoxic activity against BT-549, MCF-7, and HepG2 cell lines (IC50 = 16.82, 9.52, and 1.84 µM, respectively). Its anticancer activity was primarily due to the induction of G1/S phase arrest as discovered through cell cycle analysis of HepG2 cancer cells. Ultimately, treating HepG2 cancer cells with Pitavastatin affected significant activation of caspase-3 accompanied by down-regulation of cellular apoptotic biomarkers such as IDO, TDO, STAT3, P21, P27, IL-6, and AhR. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSC) attract an increasing amount of attention due to their unique therapeutic properties. Yet, MSC can undergo undesirable genetic and epigenetic changes during their propagation in vitro. In this study, we investigated whether polyploidy can compromise MSC oncological safety and therapeutic properties. For this purpose, we compared the impact of polyploidy on the transcriptome of cancer cells and MSC of various origins (bone marrow, placenta, and heart). First, we identified genes that are consistently ploidy-induced or ploidy-repressed through all comparisons. Then, we selected the master regulators using the protein interaction enrichment analysis (PIEA). The obtained ploidy-related gene signatures were verified using the data gained from polyploid and diploid populations of early cardiomyocytes (CARD) originating from iPSC. The multistep bioinformatic analysis applied to the cancer cells, MSC, and CARD indicated that polyploidy plays a pivotal role in driving the cell into hypertranscription. It was evident from the upregulation of gene modules implicated in housekeeping functions, stemness, unicellularity, DNA repair, and chromatin opening by means of histone acetylation operating via DNA damage associated with the NUA4/TIP60 complex. These features were complemented by the activation of the pathways implicated in centrosome maintenance and ciliogenesis and by the impairment of the pathways related to apoptosis, the circadian clock, and immunity. Overall, our findings suggest that, although polyploidy does not induce oncologic transformation of MSC, it might compromise their therapeutic properties because of global epigenetic changes and alterations in fundamental biological processes. The obtained results can contribute to the development and implementation of approaches enhancing the therapeutic properties of MSC by removing polyploid cells from the cell population. [ABSTRACT FROM AUTHOR]

Background: Cingulin (CGN) is a pivotal cytoskeletal adaptor protein located at tight junctions. This study investigates the link between CGN mutation and increased cancer susceptibility through genetic and mechanistic analyses and proposes a potential targeted therapeutic approach. Methods: In a high-cancer-density family without known pathogenic variants, we performed tumor-targeted and germline whole-genome sequencing to identify novel cancer-associated variants. Subsequently, these variants were validated in a 222 cancer patient cohort, and CGN c.3560C > T was identified as a potential cancer-risk allele. Both wild-type (WT) (c.3560C > C) and variant (c.3560C > T) were transfected into cancer cell lines and incorporated into orthotopic xenograft mice model for evaluating their effects on cancer progression. Western blot, immunofluorescence analysis, migration and invasion assays, two-dimensional gel electrophoresis with mass spectrometry, immunoprecipitation assays, and siRNA applications were used to explore the biological consequence of CGN c.3560C > T. Results: In cancer cell lines and orthotopic animal models, CGN c.3560C > T enhanced tumor progression with reduced sensitivity to oxaliplatin compared to the CGN WT. The variant induced downregulation of epithelial marker, upregulation of mesenchymal marker and transcription factor, which converged to initiate epithelial-mesenchymal transition (EMT). Proteomic analysis was conducted to investigate the elements driving EMT in CGN c.3560C > T. This exploration unveiled overexpression of IQGAP1 induced by the variant, contrasting the levels observed in CGN WT. Immunoprecipitation assay confirmed a direct interaction between CGN and IQGAP1. IQGAP1 functions as a regulator of multiple GTPases, particularly the Rho family. This overexpressed IQGAP1 was consistently associated with the activation of Rac1, as evidenced by the analysis of the cancer cell line and clinical sample harboring CGN c.3560C > T. Notably, activated Rac1 was suppressed following the downregulation of IQGAP1 by siRNA. Treatment with NSC23766, a selective inhibitor for Rac1-GEF interaction, resulted in the inactivation of Rac1. This intervention mitigated the EMT program in cancer cells carrying CGN c.3560C > T. Consistently, xenograft tumors with WT CGN showed no sensitivity to NSC23766 treatment, but NSC23766 demonstrated the capacity to attenuate tumor growth harboring c.3560C > T. Conclusions: CGN c.3560C > T leads to IQGAP1 overexpression, subsequently triggering Rac1-dependent EMT. Targeting activated Rac1 is a strategy to impede the advancement of cancers carrying this specific variant. [ABSTRACT FROM AUTHOR]

• Lamiaceae species are rich in EOs, aromatic compounds and active phytochemicals. • Phytochemical profile, antioxidant and anticancer activities of the EOs were determined in this work. • A total of thirty-seven different phytochemicals were identified by GC/MS analysis. • Anticancer activities of the EOs against NCI-N87, HepG2, MCF-7, and LNCaP cells were analyzed for the first time. • Some Lamiaceae species are potentially candidates for therapeutic purposes. The present study was aimed to determine phytochemical characterization, anticancer and antioxidant activities of essential oils obtained from 11 Lamiaceae species, including Clinopodium serpyllifolium subsp. barbatum (P.H.Davis) Bräuchler, Lavandula angustifolia Miller, Mentha × piperita L., Mentha pulegium L. , Ocimum basilicum L., Rosmarinus officinalis L., Salvia aramiensis Rech.f., Salvia fruticosa Miller, Salvia officinalis L., Satureja thymbra L., and Thymbra spicata L. taxa from Türkiye. The chemical composition of the essential oils (EOs) was identified using GC–MS, and antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′- azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) free radical scavenging methods. Furthermore, the anticancer potential of the EOs was evaluated using MTT method against human cell lines, including hepatocellular carcinoma (HepG2), gastric carcinoma (NCI-N87), breast adenocarcinoma (MCF-7), human prostate carcinoma (LNCaP clone FGC-Luc2) cancer cells, and non-cancerous human umbilical vein endothelial cells (HUVECs). The antioxidant activities of the extracts varied from 31.96 to 74.96%. Consistent with antioxidant activities, the anticancer activities of the EOs of S. officinalis, L. angustifolia , and R. officinalis were significantly higher than others. However, almost all EOs were found to inhibit cell viability and induce apoptosis of cancer cells, while the EOs at different concentrations exhibited the highest anticancer activity against NCI-N87, followed by HepG2, MCF-7, and LNCaP cancer cells, with the IC 50 value ranging from 10.98 ± 0.12 to 78.08 ± 1.21 µg/mL, respectively. Variability in phytochemical components, free radical scavenging and anticancer activities of EOs was revealed by principal component analysis (PCA) and agglomerative hierarchical clustering (AHC). The major components of the EOs distilled from Lamiaceae species were found as eucalyptol, thujone, linalool, linalyl acetate, pulegone, carvone and carvacrol at different concentrations in the EOs. Overall, it can be clearly concluded that the EO samples distilled from some medicinal plants belonging to the Lamiaceae family contain valuable phytochemical compounds and accordingly exhibit remarkable biological activities. [ABSTRACT FROM AUTHOR]

This article discusses the concept of personalized medicine in oncology, which recognizes that cancers are not all the same and that individuals may respond differently to treatments based on their genetic, environmental, and lifestyle characteristics. Precision medicine has revolutionized cancer treatment by enabling more personalized, effective, and targeted care. The article highlights specific research studies that explore personalized approaches to cancer prevention, detection, and treatment, such as correlating chemo-resistance with biological variables in endometrial cancer patients and utilizing measurable biomarkers to evaluate treatment effectiveness. The article also mentions the use of new technologies, omics sciences, and artificial intelligence in precision oncology. Overall, precision medicine has shown great potential in improving cancer treatments and providing hope for patients. [Extracted from the article]

Background: As a key enzyme in ceramide synthesis, longevity assurance homologue 2 (LASS2) has been indicated to act as a tumour suppressor in a variety of cancers. Ferroptosis is involved in a variety of tumour processes; however, the role of LASS2 in regulating ferroptosis has yet to be explored. This article explores the potential underlying mechanisms involved. Methods: Bioinformatics tools and immunohistochemical staining were used to evaluate LASS2 expression, and the results were analysed in relation to overall survival and clinical association in multiple cancers. Coimmunoprecipitation-coupled liquid chromatography-mass spectrometry (co-IP LC-MS) was performed to identify potential LASS2-interacting proteins in thyroid, breast, and liver cancer cell lines. Transcriptomics, proteomics and metabolomics analyses of multiple cancer cell types were performed using MS or LC–MS to further explore the underlying mechanisms involved. Among these tumour cells, the common LASS2 interaction partner transferrin receptor (TFRC) was analysed by protein–protein docking and validated by coimmunoprecipitation western blot, immunofluorescence, and proximity ligation assays. Then, we performed experiments in which tumour cells were treated with Fer-1 or erastin or left untreated, with or without inducing LASS2 overexpression, and assessed the molecular biological and cellular functions by corresponding analyses. Results: Low LASS2 expression is correlated with adverse clinical characteristic and poor prognosis in patients with thyroid cancer, breast cancer or HCC. Multiomics analyses revealed significant changes in the ferroptosis signalling pathway, iron ion transport and iron homeostasis. Our in vitro experiments revealed that LASS2 overexpression regulated ferroptosis status in these tumour cells by affecting iron homeostasis, which in turn inhibited tumour migration, invasion and EMT. In addition, LASS2 overexpression reversed the changes in tumour cell metastasis induced by either Fer-1 or erastin. Mechanistically, LASS2 interacts directly with TFRC to regulate iron homeostasis in these tumour cells. Conclusions: In summary, our study reveals for the first time that LASS2 can inhibit tumour cell metastasis by interacting with TFRC to regulate iron metabolism and influence ferroptosis status in thyroid, breast, and liver cancer cells, these results suggest potential universal therapeutic targets for the treatment of these cancers. [ABSTRACT FROM AUTHOR]

Background: AlkB homolog 1, histone H2A dioxygenase (ALKBH1), a crucial enzyme involved in RNA demethylation in humans, plays a significant role in various cellular processes. While its role in tumor progression is well-established, its specific contribution to stomach adenocarcinoma (STAD) remains elusive. This study seeks to explore the clinical and pathological relevance of ALKBH1, its impact on the tumor immune microenvironment, and its potential for precision oncology in STAD. Methods: We adopted a comprehensive multi-omics approach to identify ALKBH1 as an potential diagnostic biomarker for STAD, demonstrating its association with advanced clinical stages and reduced overall survival rates. Our analysis involved the utilization of publicly available datasets from GEO and TCGA. We identified differentially expressed genes in STAD and scrutinized their relationships with immune gene expression, overall survival, tumor stage, gene mutation profiles, and infiltrating immune cells. Moreover, we employed spatial transcriptomics to investigate ALKBH1 expression across distinct regions of STAD. Additionally, we conducted spatial transcriptomic and single-cell RNA-sequencing analyses to elucidate the correlation between ALKBH1 expression and immune cell populations. Our findings were validated through immunohistochemistry and bioinformatics on 60 STAD patient samples. Results: Our study unveiled crucial gene regulators in STAD linked with genetic variations, deletions, and the tumor microenvironment. Mutations in these regulators demonstrated a positive association with distinct immune cell populations across six immune datasets, exerting a substantial influence on immune cell infiltration in STAD. Furthermore, we established a connection between elevated ALKBH1 expression and macrophage infiltration in STAD. Pharmacogenomic analysis of gastric cancer cell lines further indicated that ALKBH1 inactivation correlated with heightened sensitivity to specific small-molecule drugs. Conclusion: In conclusion, our study highlights the potential role of ALKBH1 alterations in the advancement of STAD, shedding light on novel diagnostic and prognostic applications of ALKBH1 in this context. We underscore the significance of ALKBH1 within the tumor immune microenvironment, suggesting its utility as a precision medicine tool and for drug screening in the management of STAD. [ABSTRACT FROM AUTHOR]

The overexpression of the immunoinhibitory receptor programmed death-1 (PD1) on T-cells is involved in immune evasion in cancer. The use of anti-PD-1/PDL-1 strategy has deeply changed the therapies of cancers and patient survival. However, their efficacy diverges greatly along with tumor type and patient populations. Thereby, novel treatments are needed to interfere with the anti-tumoral immune responses and propose an adjunct therapy. In the current study, we found that the antifungal drug Sulconazole (SCZ) inhibits PD-1 expression on activated PBMCs and T cells at the RNA and protein levels. SCZ repressed NF-κB and calcium signaling, both, involved in the induction of PD-1. Further analysis revealed cancer cells treatment with SCZ inhibited their proliferation, and migration and ability to mediate tumor growth in zebrafish embryos. SCZ found also to inhibit calcium mobilization in cancer cells. These results suggest the SCZ therapeutic potential used alone or as adjunct strategy to prevent T-cell exhaustion and promotes cancer cell malignant phenotype repression in order to improve tumor eradication. [ABSTRACT FROM AUTHOR]

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced, and 599 candidate genes exhibited differential expression between DSG3-overexpressing and matched control lines, with 12 genes highly significantly differentially expressed, including 9 upregulated and 3 downregulated. Genes with known implications in cancer, such as MMP-13, KRT84, OLFM4, GJA1, AMOT and ADAMTS1, were strongly linked to DSG3 overexpression. Gene ontology analysis indicated that the DSG3-associated candidate gene products participate in crucial cellular processes such as junction assembly, focal adhesion, extracellular matrix formation, intermediate filament organisation and keratinocyte differentiation. Validation of RNA-Seq was performed through RT-qPCR, Western blotting and immunofluorescence analyses. Furthermore, using transmission electron microscopy, we meticulously examined desmosome morphology and revealed a slightly immature desmosome structure in DSG3-overexpressing cells compared to controls. No changes in desmosome frequency and diameter were observed between the two conditions. This study underscores intricate and multifaceted alterations associated with DSG3 in oral squamous carcinoma cells, implying a potential oncogenic role of this gene in biological processes that enable cell communication, motility and survival. [ABSTRACT FROM AUTHOR]

Single cell computational analysis has emerged as a powerful tool in the field of oncology, enabling researchers to decipher the complex cellular heterogeneity that characterizes cancer. By leveraging computational algorithms and bioinformatics approaches, this methodology provides insights into the underlying genetic, epigenetic and transcriptomic variations among individual cancer cells. In this paper, we present a comprehensive overview of single cell computational analysis in oncology, discussing the key computational techniques employed for data processing, analysis, and interpretation. We explore the challenges associated with single cell data, including data quality control, normalization, dimensionality reduction, clustering, and trajectory inference. Furthermore, we highlight the applications of single cell computational analysis, including the identification of novel cell states, the characterization of tumor subtypes, the discovery of biomarkers, and the prediction of therapy response. Finally, we address the future directions and potential advancements in the field, including the development of machine learning and deep learning approaches for single cell analysis. Overall, this paper aims to provide a roadmap for researchers interested in leveraging computational methods to unlock the full potential of single cell analysis in understanding cancer biology with the goal of advancing precision oncology. For this purpose, we also include a notebook that instructs on how to apply the recommended tools in the Preprocessing and Quality Control section. [ABSTRACT FROM AUTHOR]

Extracted from the roots of Plumbago zeylanica L., plumbagin is a natural naphthoquinone with potential as an anticancer compound. However, no studies have investigated its impact on LoVo (colon cancer) cells, and the specific mechanisms by which plumbagin exerts its anticancer effects remain to be established. The anticancer potential of plumbagin against LoVo cells was evaluated using a battery of assays, including MTT assay, clone formation assay, transwell chamber invasion assay, and wound‐curing assay. Cell cycle analysis and cell apoptosis analysis were conducted to break down the anticancer impact of plumbagin on LoVo cells. A label‐free proteomics technology was employed to investigate alterations in protein expression in LoVo cells treated with plumbagin. Our investigation indicated that plumbagin markedly inhibited the LoVo cells proliferation, and induced the apoptosis in LoVo cells, simultaneously induced G0/G1 phase cell cycle arrest. The LC‐MS/MS proteomics assay revealed 78 proteins that were differentially expressed upon treatment with plumbagin. Bioinformatics and functional analyses indicated that these proteins were predominantly involved in protein synthesis and translation. Our findings revealed that multiple mechanisms are involved in the anticancer activity of plumbagin against LoVo cells, resulting in decreased cell viability. Proteomic analysis suggests that plumbagin may impede protein synthesis by reducing the expression of eukaryotic initiation factors. Our findings demonstrate that plumbagin exerts its anticancer activity against LoVo cells through multiple mechanisms, including inhibition of cell proliferation, induction of apoptosis, cell cycle arrest, and disruption of protein synthesis. These results provide new insights into the therapeutic potential of plumbagin for colon cancer treatment. [ABSTRACT FROM AUTHOR]

Recently, research incorporating the benefits of deep learning in the application of cancer cell classification and analysis has been actively conducted. In this paper, we investigated examples of binary-class classification and multi-class classification of cancer cell image data of commonly occurring types of cancer worldwide, such as cervical cancer, breast cancer, lung cancer, and colon cancer, using convolutional neural networks (CNNs) models. For instance, some studies explored the utilization of transfer learning, leveraging a pre-trained CNN model is used as a starting point for additional training on a specific cancer cell dataset. Cancer cells have irregular and abnormal growth, making accurate classification challenging. The application of deep learning techniques, such as CNN, for cancer cell classification has been able to solve these complex analysis problems and enable fast cancer cell classification results, leading to early detection of cancer. Indeed, most of the studies in this paper achieved high performance using CNN models, and this approach enables faster and more accurate confirmation of cancer cell classification results, leading to early detection of cancer. This shows the current trend of applying deep learning in the application of cancer cell classification and demonstrates the significant potential of deep learning to contribute to cancer research. Overall, we provide an overview of the current trend of applying deep learning in the field of cancer cell classification and expect that deep learning will open the way for more effective cancer diagnosis and treatment in the future. [ABSTRACT FROM AUTHOR]

Objective(s): The present study evaluated the protein-based analysis to unravel the role and mechanism behind the Dendropthae falcata plant extract treatment in breast cancer cells. Materials and Methods: The protein sample was extracted from the cancer cells after treatment with the plant extract and subjected to two-dimensional electrophoresis for protein separation. Further, the proteins that were differentially regulated among the samples which were treated and non-treated were selected and processed further for protein identification using a tandem mass spectrometry approach. Results: Using these strategies, we identified 16 potential candidates which were showing remarkable changes in treated samples. All the candidates were analyzed further for gene ontology analysis, and it was observed that all proteins were involved in multiple pathways pertaining to the carcinogenesis process. Specifically, apoptotic pathway proteins including BAD, BIK, BID, CASP8, MCL1, BCL2, and BAK1 were highly impacted by treatment with D. falcata plant extract. All these protein hits were further taken for validation experiments using RT PCR analysis. Conclusion: Initiation of these apoptotic proteins by D. falcata plant extract treatment in breast cancer cells shows a positive direction toward nature-based alternative medicine. [ABSTRACT FROM AUTHOR]