human renal cancer cell lines a498  (ATCC)

Journal: bioRxiv

Article Title: Fibroblast-derived Collagen VI shapes the structure and function of the tumor-immune microenvironment in clear cell renal cell carcinoma

doi: 10.64898/2026.03.19.712351

Figure Lengend Snippet: (A) Schematic depicting cancer cell reseeding on TK173-synthesized CDMs and further functional workup (Created in BioRender. Schell, C. (2026) https://BioRender.com/x5sqb3y ). (B) Volcano plot of RNA sequencing analysis depicting differentially expressed genes of A498 cancer cells after reseeding on TK173-synthesized CDMs (COL6-containing CDM or depleted CDM; red dots indicate significantly regulated genes with adjusted p<0.05 and log 2 fold change (FC) > |0.5|). (C) Gene ontology (GO) gene set enrichment analysis of GO-terms of the molecular function category. (D) Volcano plot analysis for filtered matrisome genes. (E) IF staining for Nucleus (blue), F-actin (white), FN (green) and COL6 (violet) of A498 cells reseeded on shCTRL- and shCOL6-CDMs. (F) Scanning electron microscopy of A498 cells interacting with CDMs (yellow dotted boxes highlight areas of zoom in). (G&H) Morphometric analysis of reseeded cells (cell area and perimeter; Violin plots of individual cells of N=3 independent experiments, Mann-WhitneyLULtest). (I) Quantification of confluence of A498 cells growing on shCTRL- and shCOL6-CDMs (N=4 independent experiments, unpaired t test). (J&K) Representative phase-contrast images with DNA staining (blue) of A498 cells after cultivation for 3 days. Dot plot depicting quantification of A498 cells per cm² grown on CDMs (N=4 independent experiments, unpaired t test). (L&M) Representative IF staining of ccRCC patient samples for DNA (blue), PAX8 (red), KI-67 (green) and COL6 (violet) was used to classify and mask (bottom image) for proliferating tumor cells (orange), non-proliferative tumor cells (grey), as well as COL6 (violet). Violin plot depicting quantification of the distance of non-proliferative and proliferating tumor cells to COL6 (Mann-WhitneyLULtest). (N) Heatmap depicting Spearman’s correlation analysis of indicated proliferative cell state marker genes with COL6 , SERPINE1 and SERPINE2 genes in the ccRCC TCGA and CPTAC cohorts. (O&P) Spatial transcriptomics analysis of ccRCCs for spatial co-occurrence of COL6 , SERPINE1 , SERPINE2 and proliferative cell state marker genes. (O) Exemplary spatial mapping of the co-occurrence score on one sample (PD47512) and spatial mapping for the COL6, SERPINE and proliferative gene scores. (P) Correlation matrix of the spatial correlation between the COL6, SERPINE and proliferative gene sets (N=5 patients, Spearman’s correlation). Bars indicate mean and S.E.M in dot plots or median and quartiles in violin plots; ∗ – p < 0.05, ∗∗ – p < 0.01 and ∗∗∗∗ – p < 0.0001.

Article Snippet: Human renal cancer cell lines A498 (HTB-44, ATCC,) and 786-O (CRL-1932, ATCC) as well as human renal fibroblasts TK173 (RRID: CVCL_C8FA) and human renal proximal tubule epithelial cells (RPTEC/TERT1, ATCC, CRL-4031) were used and obtained as recently described .

Techniques: Synthesized, Functional Assay, RNA Sequencing, Staining, Electron Microscopy, Marker, Spatial Transcriptomics

Journal: Molecular Oncology

Article Title: Obesity alters the fitness of peritumoral adipose tissue, exacerbating tumor invasiveness in renal cancer through the induction of ADAM12 and CYP1B1

doi: 10.1002/1878-0261.13782

Figure Lengend Snippet: Adipose peritumoral tissue affects the expression of inflammatory genes by renal cell carcinoma (RCC) cells. (A) Relative mRNA expression of the different tumor suppressor genes in the healthy and tumor kidney tissues of RCC patients ( n = 10), including VHL , PBRM1 , SETD2 , and BAP1 , was measured by real‐time PCR. Cyclophilin‐A was used as an internal control. Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by Student's t ‐test. (B) Schematic figure illustrating the protocol for differentiation of adipocytes from peritumoral adipose tissue (AT) of lean, overweight, and obese RCC patients. (C) Relative mRNA expression of the leptin and adiponectin of the differentiated adipocytes among the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. (D) Schematic figure of the conditioned medium (CM) experiment, showing RCC cells treated with conditioned media (CMs) of lean, overweight, and obese differentiated adipocytes. (E) Relative mRNA expression of the IL6 , CXCR4 , SDF1 , and BAFFR of the A498 cells treated with CMs collected from differentiated adipocytes derived from the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The human renal cancer cell line (A498) was grown in DMEM (Microgem); supplemented with 10% FBS, 1% Pen/Strep, and 1% L‐Glutamine, at 37 °C in 5% CO 2 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Derivative Assay

Journal: American Journal of Cancer Research

Article Title: Evaluation of the role of downregulation of SNF5/INI1 core subunit of SWI/SNF complex in clear cell renal cell carcinoma development

doi:

Figure Lengend Snippet: INI1 influences CXCL12/CXCR4/CXCR7 chemokine axis. A. INI1-FLAG overexpression in A498 cells, which correspond with ccRCC grade 2. As a loading control membrane stained with TCE (stain free) was used. B. Western blot with anti-INI1 antibody demonstrating increase of INI1 abundance in A498 cell line overexpressing INI1 protein. As a loading control membrane stained with TCE (stain free) was used. C. qRT-PCR analysis for CXCR4, CXCR7 and SMARCB1 (endogenous transcript) genes expression in A498 cell lines overexpressing INI1 protein. D. Model describing self-regulation of the SMARCB1 gene by INI1 protein and the interdependence of INI1, AR pathway and CXCL12/CXCR4/CXCR7 axis.

Article Snippet: A498 human renal cancer cell line (obtained from ATCC) was grown in RPMI 1640 (Biowest) with 10% Fetal Bovine Serum (FBS) (Biowest) and 1% penicillin/streptomycin (Gibco) in humidifier incubator (37°C and 5% CO 2 ) according to manufacturer instruction.

Techniques: Over Expression, Control, Membrane, Staining, Western Blot, Quantitative RT-PCR, Expressing

Journal: American Journal of Cancer Research

Article Title: Evaluation of the role of downregulation of SNF5/INI1 core subunit of SWI/SNF complex in clear cell renal cell carcinoma development

doi:

Figure Lengend Snippet: INI1 influences CXCL12/CXCR4/CXCR7 chemokine axis. A. INI1-FLAG overexpression in A498 cells, which correspond with ccRCC grade 2. As a loading control membrane stained with TCE (stain free) was used. B. Western blot with anti-INI1 antibody demonstrating increase of INI1 abundance in A498 cell line overexpressing INI1 protein. As a loading control membrane stained with TCE (stain free) was used. C. qRT-PCR analysis for CXCR4, CXCR7 and SMARCB1 (endogenous transcript) genes expression in A498 cell lines overexpressing INI1 protein. D. Model describing self-regulation of the SMARCB1 gene by INI1 protein and the interdependence of INI1, AR pathway and CXCL12/CXCR4/CXCR7 axis.

Article Snippet: Cell culture and transfection A498 human renal cancer cell line (obtained from ATCC) was grown in RPMI 1640 (Biowest) with 10% Fetal Bovine Serum (FBS) (Biowest) and 1% penicillin/streptomycin (Gibco) in humidifier incubator (37°C and 5% CO 2 ) according to manufacturer instruction.

Techniques: Over Expression, Control, Membrane, Staining, Western Blot, Quantitative RT-PCR, Expressing