human ccrcc cell lines caki 2 htb 47  (ATCC)

Journal: Oncology Reports

Article Title: Mechanistic study of MUC3A in promoting progression of clear cell renal cell carcinoma via the JAK-STAT pathway

doi: 10.3892/or.2026.9119

Figure Lengend Snippet: MUC3A is aberrantly upregulated in ccRCC and associated with poor prognosis. (A) Pan-cancer analysis of MUC3A expression across multiple tumor types based on TCGA data, generated using the GEPIA2 platform. Gene expression values are presented as log2(TPM + 1). (B) Differential expression of MUC3A in tumor and normal samples from the TCGA-KIRC cohort (523 tumor samples vs. 72 normal samples). *P<0.05. (C) Western blotting of MUC3A protein expression in HK-2 and RPTEC/TERT1 non-malignant renal epithelial cell lines and ccRCC cell lines (CAKI-1, OSRC-2, 786-O and ACHN). GAPDH was used as a loading control. (D) Western blotting of MUC3A knockdown efficiency in 786-O and OSRC-2 cells following transient transfection with three independent siRNAs targeting MUC3A. si-2 was selected for subsequent functional experiments due to its superior knockdown efficiency. (E) Kaplan-Meier OS analysis of ccRCC patients stratified into high- and low-MUC3A expression groups using the GEPIA2 platform. (F) Kaplan-Meier DFS analysis of ccRCC patients based on MUC3A expression levels. Data are derived from TCGA unless otherwise indicated. MUC3A, mucin 3A; ccRCC, clear cell renal cell carcinoma; TCGA, The Cancer Genome Atlas; KIRC, kidney renal clear cell carcinoma; GEPIA2, Gene Expression Profiling Interactive Analysis 2; TPM, transcripts per million; KIRC, kidney renal clear cell carcinoma; OS, overall survival; DFS, disease-free survival; siRNA, small interfering RNA; si-MUC3A, MUC3A-targeting siRNA; si-Ctrl, non-targeting control siRNA; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; si-1/si-2/si-3, three independent siRNAs targeting MUC3A.

Article Snippet: Human ccRCC cell lines 786-O (cat. no. CL-0010), OSRC-2 (cat. no. CL-0177), Caki-1 (cat. no. CL-0052) and ACHN (cat. no. CL-0021), as well as non-malignant renal epithelial cells HK-2 (cat. no. CL-0109; all from Procell Life Science & Technology Co., Ltd.) and RPTEC/TERT1 (cat. no. CRL-4031; American Type Culture Collection), were used in the present study.

Techniques: Expressing, Generated, Gene Expression, Quantitative Proteomics, Western Blot, Control, Knockdown, Transfection, Functional Assay, Derivative Assay, Small Interfering RNA

Journal: Oncology Reports

Article Title: Mechanistic study of MUC3A in promoting progression of clear cell renal cell carcinoma via the JAK-STAT pathway

doi: 10.3892/or.2026.9119

Figure Lengend Snippet: MUC3A knockdown suppresses proliferation and promotes apoptosis in ccRCC cells. (A and B) Cell proliferation of 786-O and OSRC-2 cells following transfection with si-MUC3A or si-Ctrl, as assessed by CCK-8 assays at the indicated time points. (C and D) Colony formation assays showing the clonogenic capacity of 786-O and OSRC-2 cells following MUC3A knockdown. Representative images and quantitative analysis are shown. (E and F) Flow cytometric analysis of apoptosis in 786-O and OSRC-2 cells using Annexin V-FITC/PI staining following MUC3A silencing. Representative dot plots and corresponding quantitative results are presented. All experiments were performed with at least three independent biological replicates (n≥3). Data are presented as the mean ± SD. Statistical significance was determined using a two-tailed unpaired Student's t-test. *P<0.05, **P<0.01 and ***P<0.001. MUC3A, mucin 3A; ccRCC, clear cell renal cell carcinoma; CCK-8, Cell Counting Kit-8; si-MUC3A, MUC3A-targeting siRNA; si-Ctrl, non-targeting control siRNA; PI, propidium iodide; SD, standard deviation; ns, not significant.

Article Snippet: Human ccRCC cell lines 786-O (cat. no. CL-0010), OSRC-2 (cat. no. CL-0177), Caki-1 (cat. no. CL-0052) and ACHN (cat. no. CL-0021), as well as non-malignant renal epithelial cells HK-2 (cat. no. CL-0109; all from Procell Life Science & Technology Co., Ltd.) and RPTEC/TERT1 (cat. no. CRL-4031; American Type Culture Collection), were used in the present study.

Techniques: Knockdown, Transfection, CCK-8 Assay, Staining, Two Tailed Test, Cell Counting, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Mechanistic study of MUC3A in promoting progression of clear cell renal cell carcinoma via the JAK-STAT pathway

doi: 10.3892/or.2026.9119

Figure Lengend Snippet: MUC3A knockdown inhibits the migration and invasion of ccRCC cells. (A) Transwell migration assays showing the migratory capacity of 786-O and OSRC-2 cells following MUC3A knockdown. Quantitative analysis is shown on the right. (B) Transwell invasion assays performed using Matrigel ® -coated chambers to assess the invasive potential of ccRCC cells after MUC3A silencing. (C) Wound healing assays demonstrating delayed wound closure in 786-O and OSRC-2 cells transfected with si-MUC3A compared with si-Ctrl at 24 h. Representative images and quantitative analyses are shown. Scale bar, 100 µm. All experiments were conducted with at least three independent biological replicates. Data are expressed as the mean ± SD. **P<0.01 and ***P<0.001. MUC3A, mucin 3A; ccRCC, clear cell renal cell carcinoma; si-MUC3A, MUC3A-targeting siRNA; si-Ctrl, non-targeting control siRNA; SD, standard deviation.

Article Snippet: Human ccRCC cell lines 786-O (cat. no. CL-0010), OSRC-2 (cat. no. CL-0177), Caki-1 (cat. no. CL-0052) and ACHN (cat. no. CL-0021), as well as non-malignant renal epithelial cells HK-2 (cat. no. CL-0109; all from Procell Life Science & Technology Co., Ltd.) and RPTEC/TERT1 (cat. no. CRL-4031; American Type Culture Collection), were used in the present study.

Techniques: Knockdown, Migration, Transfection, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Mechanistic study of MUC3A in promoting progression of clear cell renal cell carcinoma via the JAK-STAT pathway

doi: 10.3892/or.2026.9119

Figure Lengend Snippet: MUC3A is associated with the activation of the JAK-STAT signaling pathway in ccRCC. (A) KEGG pathway enrichment analysis of genes associated with MUC3A expression based on TCGA-KIRC transcriptomic data. (B) GSEA showing significant enrichment of the JAK-STAT signaling pathway in ccRCC samples with a high MUC3A expression. (C and D) Western blotting of total and phosphorylated JAK1, JAK2 and STAT3 in 786-O and OSRC-2 cells following MUC3A knockdown. (E) Western blotting showing that STAT3 activation by Colivelin TFA restores p-STAT3 levels in si-MUC3A-transfected 786-O and OSRC-2 cells, accompanied by increased Bcl-2 and decreased cleaved caspase-3 expression. (F) Western blotting of apoptosis-related proteins Bcl-2 and cleaved caspase-3 following MUC3A silencing. GAPDH served as a loading control. All western blotting experiments were repeated independently at least three times. MUC3A, mucin 3A; JAK, Janus kinase; STAT, signal transducer and activator of transcription; ccRCC, clear cell renal cell carcinoma; KEGG, Kyoto Encyclopedia of Genes and Genomes; TCGA, The Cancer Genome Atlas; KIRC, kidney renal clear cell carcinoma; GSEA, gene set enrichment analysis; TFA, trifluoroacetate; p-, phosphorylated.

Article Snippet: Human ccRCC cell lines 786-O (cat. no. CL-0010), OSRC-2 (cat. no. CL-0177), Caki-1 (cat. no. CL-0052) and ACHN (cat. no. CL-0021), as well as non-malignant renal epithelial cells HK-2 (cat. no. CL-0109; all from Procell Life Science & Technology Co., Ltd.) and RPTEC/TERT1 (cat. no. CRL-4031; American Type Culture Collection), were used in the present study.

Techniques: Activation Assay, Expressing, Western Blot, Knockdown, Transfection, Control

Journal: Oncology Reports

Article Title: Mechanistic study of MUC3A in promoting progression of clear cell renal cell carcinoma via the JAK-STAT pathway

doi: 10.3892/or.2026.9119

Figure Lengend Snippet: STAT3 activation partially rescues the effects of MUC3A knockdown in ccRCC cells. (A and B) CCK-8 assays showing that treatment with the STAT3 agonist Colivelin TFA partially restored the proliferation of 786-O and OSRC-2 cells following MUC3A knockdown. (C-F) Flow cytometric analysis demonstrating that Colivelin TFA treatment reverses the apoptosis-promoting effect induced by MUC3A silencing in ccRCC cells. Data are presented as the mean ± SD from at least three independent biological replicates. Statistical significance was assessed using Student's t-test or one-way ANOVA as appropriate. *P<0.05, **P<0.01 and ***P<0.001. STAT3, signal transducer and activator of transcription 3; TFA, trifluoroacetate; MUC3A, mucin 3A; CCK-8, Cell Counting Kit-8; si-MUC3A, MUC3A-targeting siRNA; si-Ctrl, non-targeting control siRNA; SD, standard deviation; ANOVA, analysis of variance.

Article Snippet: Human ccRCC cell lines 786-O (cat. no. CL-0010), OSRC-2 (cat. no. CL-0177), Caki-1 (cat. no. CL-0052) and ACHN (cat. no. CL-0021), as well as non-malignant renal epithelial cells HK-2 (cat. no. CL-0109; all from Procell Life Science & Technology Co., Ltd.) and RPTEC/TERT1 (cat. no. CRL-4031; American Type Culture Collection), were used in the present study.

Techniques: Activation Assay, Knockdown, CCK-8 Assay, Cell Counting, Control, Standard Deviation

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: The process of identifying differentially expressed IMRGs and molecular subtypes in ccRCC: ( A ) IMRGs that were differentially expressed were denoted by red dots for upregulation and blue dots for downregulation. ( B ) Heatmaps were used to visually represent the top differentially expressed genes. ( C ) A heatmap of the nsNMF consensus matrix was generated to classify ccRCC into two molecular subtypes. ( D ) A PCA plot was applied to show significant differences between clusters. ( E ) The gene expression heatmap shows how the identified IMRGs were expressed across the two molecular subtypes. ( F , G ) In order to make a comparison between the two molecular subtypes, the researcher employed the Kaplan–Meier curve to assess and contrast the OS and PFS.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Generated, Gene Expression, Comparison

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Comparison of genomic alteration landscapes between the two molecular subtypes. ( A ) Oncoplot demonstrated the 30 most frequently mutated genes in Cluster 1. ( B ) Oncoplot demonstrated the 30 most frequently mutated genes in Cluster 2. ( C ) Heatmap illustrating the co-mutated states of the commonly mutated genes in Cluster 1. ( D ) Heatmap illustrating the co-mutated states of the commonly mutated genes in Cluster 2. ( E ) The boxplot illustrates the distinct tumor mutation frequencies between Cluster 1 and Cluster 2. ( F ) The Kaplan–Meier curve shows the overall survival rates of patients with high and low tumor mutation burdens. ( G ) Multivariate Cox regression analysis of tumor mutation burden (TMB) and immunometabolic clusters. ( H ) Kaplan–Meier survival curves for ccRCC patients stratified by both TMB status (high vs. low) and immunometabolic clusters (C1 vs. C2).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Comparison, Mutagenesis

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Assessment and confirmation of the predictive performance of the signature in ccRCC. ( A – C ) Scatter plots illustrating the survival status and IMI scores of ccRCC patients in the TCGA training group ( A ), the TCGA testing group ( B ), and the E-MATB-1980 external validation group ( C ). ( D – F ) Kaplan–Meier curves displaying the overall survival situation per IMI scores of the high-IMI group and low-IMI group in the TCGA training group ( D ), the TCGA testing group ( E ), and the E-MATB-1980 external validation group ( F ). ( G – I ) ROC curves demonstrating the predictive performance of IMI with AUC values for 1-year, 3-year, and 5-year OS in ccRCC patients from the TCGA training group ( G ), the TCGA testing group ( H ), and the E-MATB-1980 external validation group ( I ).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Biomarker Discovery

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Identification of expression trends of nine IMRGs. ( A ) Differences in signature gene expression between high and low IMI groups in the TCGA database. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ( B ) Differences in signature gene expression between normal kidney tissue samples and ccRCC samples in the TCGA database. ( C – K ) The relative expression levels of signature genes between three ccRCC cell lines (786-O, A498, ACHN) and normal renal tubular epithelial cells, HK2. ( L ) The IHC images compared the expression levels of four signature genes between normal renal tissue samples and ccRCC samples in the HPA database ( https://www.proteinatlas.org , accessed on 1 January 2024).

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Expressing, Gene Expression

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: Verification of UCN promoting proliferation, migration, and invasion of ccRCC. ( A ) Knockdown of the UCN gene in 786-O and ACHN cells, relative mRNA levels in the negative control (NC) group and three siRNA knockdown groups, respectively. **** p < 0.0001 ( B ) The knockdown effect of three siRNAs on the UCN gene at the protein level in two cell lines. The uncropped blots are shown in . ( C ) The proliferation curves of CCK8 in the control group and the knockdown groups of the two cell lines. Any siRNA group has significant statistical differences from the NC group. ( D , E ) Wound-healing assays in control and knockdown groups of the two cell lines. ( F , G ) Transwell invasion assays in control and knockdown groups of the two cell lines.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Migration, Knockdown, Negative Control, Control

Journal: Cancers

Article Title: An Integrated Immunometabolic Signature Predicts Prognosis and Immunotherapy Response in ccRCC and Identifies UCN -Mediated Immune Evasion as a Therapeutic Vulnerability: Evidence from In Vitro and In Vivo Studies

doi: 10.3390/cancers18091373

Figure Lengend Snippet: UCN regulates the immune microenvironment and promotes ccRCC progression. ( A ) Schematic illustration of the mouse xenograft tumor model experimental design. ( B – E ) Tumor growth analyses demonstrate reduced tumor volume and weight across different experimental groups, with notable suppression in sh UCN +IgG2a and sh UCN +PD-1 groups. ( F ) Gating strategy for tumor-infiltrating lymphocytes. Representative flow plots showing the identification of Live/CD45+ cells, T cells (CD3+), CD4+ and CD8+ subsets, as well as Tregs and PD-1+ cells. ( G ) Flow cytometry analysis unveils substantial alterations in immune cell subsets in the tumor immune microenvironment. ( H , I ) Representative mIHC staining of tumors (green: CD8, red: Foxp3, blue: DAPI; scale bar, 50 μm.) ( I ) The column diagram showing the counts of spots with CD8+ T cells and Tregs in tumor slides. Data presented as Mean ± SEM. One-way ANOVA was used in ( E , G , I ). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human ccRCC cell line 786-O (Accession Number: CVCL_1051) and mouse ccRCC cell line Renca (CVCL_2174) were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia) and cultured in RPMI 1640 medium (Procell, Wuhan, China) containing 10% fetal bovine serum (Procell, China) and Penicillin–Streptomycin (Procell, China).

Techniques: Flow Cytometry, Staining