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: 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: Discover Oncology

Article Title: Comprehensive analysis of PANoptosis-related molecular subtypes and prognostic model development of clear cell renal cell carcinoma

doi: 10.1007/s12672-026-04561-9

Figure Lengend Snippet: Knockdown of RBCK1 significantly inhibits ccRCC cell proliferation and migration. A Differential expression analysis of RBCK1 between normal and tumor groups. B–D DSS, PFI, and OS survival curves for low- and high-RBCK1 expression subgroups. E Protein expression levels of RBCK1 in normal and tumor groups based on the CPTAC database. F Protein expression levels of RBCK1 in HK2 cells and 786-O cells with quantitative analysis (n = 3). G Protein expression levels of RBCK1 in HK2 cells and 769-P cells with quantitative analysis (n = 3). H, I Colony formation assay and Transwell migration assay in 786-O cells (n = 3). J, K Colony formation assay and Transwell migration assay in 769-P cells (n = 3). L, M CCK-8 assay showing cell viability at different time points in 786-O and 769-P cells (n = 3). Data are presented as mean ± SD; significance: * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human renal proximal tubular epithelial cells HK2, human ccRCC 786-O cells, and human ccRCC 769-P cells were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Knockdown, Migration, Quantitative Proteomics, Expressing, Colony Assay, Transwell Migration Assay, CCK-8 Assay