human ccrcc cell lines 786 o  (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: 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: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Identification and Validation of Hypoxia and Angiogenesis-Related Differentially Expressed Hub Genes (hHA-DEGs). ( A ) Correlation between hypoxia and angiogenesis gene sets in tumor samples from TCGA. ( B ) Expression scores of hypoxia and angiogenesis gene sets in ccRCC tumor tissues versus non-tumor tissues, based on single-cell dataset GSE14526 . ( C ) WGCNA of modules associated with tissue type, vascular proliferation, and hypoxia gene sets. Eighteen modules correlated with the three phenotypes were identified. ( D ) By intersecting WCGNA genes with differentially expressed genes of TCGA and hypoxia-angiogenesis gene set, 13 key differentially expressed genes (hHA-DEGs) were identified.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Biomarker Discovery, Expressing, Single Cell

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Construction of a prognostic model related to hypoxia-angiogenesis. ( A ) The CI index of the top 18 algorithms for both the training and validation sets. ( B ) ROC curve and Kaplan Meier curve for the prognostic model in the TCGA-KIRC training set. ( C ) ROC curve and Kaplan Meier curve for the prognostic model in the E-MTAB-1980 validation set. ( D ) Nome plot displaying the prognostic significance of the six selected hHA-DEGs within the TCGA ccRCC cohorts. ( E ) Prognostic calibration curve assessing the agreement between predicted and actual risks for the six hHA-DEGs within the TCGA ccRCC cohor.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Biomarker Discovery

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Single cell analysis and RT-PCR validation. ( A ) Violin plots depicting the expression levels of RPL36A, AK3, LGALS1, TIMP1, and VIM across various cell types within ccRCC tissues, based on the single-cell dataset GSE14526 . ( B ) Comparative analysis of RNA expression levels of RPL36A, AK3, LGALS1, TIMP1, and VIM between normal and tumor tissues from ccRCC patients. ( C ) Forest plots summarizing the univariate and multivariate Cox proportional hazards regression analyses of the prognostic value of RPL36A, AK3, LGALS1, TIMP1, and VIM in the TCGA ccRCC cohort. The red box highlights AK3, which was identified as an independent protective prognostic factor in the multivariate analysis. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Single-cell Analysis, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Expressing, Single Cell, RNA Expression

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Bioinformatics Analysis of AK3 Expression, Clinical Features, and Prognosis in ccRCC. ( A ) Boxplot analysis of AK3 mRNA expression levels across pan-cancers in TCGA. ( B ) Paired analysis of AK3 mRNA expression in normal versus tumor tissues in ccRCC. ( C ) Receiver Operating Characteristic (ROC) curve illustrating the diagnostic accuracy of AK3 expression for ccRCC. ( D ) Boxplot of AK3 protein expression levels in normal and primary tumor tissues from the CPTAC data. ( E ) Boxplots depicting AK3 expression correlation with clinical features, including pathologic M stage, histologic grade, and gender. ( F ) Kaplan-Meier survival curves showing the relationship between AK3 expression and OS, DSS and PFI in ccRCC patients. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Expressing, Diagnostic Assay

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Validation of AK3 Expression Levels in ccRCC Tissue Samples and Cell Lines. ( A ) Western blot analysis of AK3 protein expression in normal control (NC) and tumor tissues from ccRCC. ( B ) Immunohistochemical (IHC) analysis of AK3 expression in a tissue microarray containing normal (N) and tumor (T) samples from ccRCC patients. H-Score quantification of AK3 expression demonstrates significant downregulation in tumor tissues compared to normal tissues. ( C ) Western blot analysis of AK3 protein expression in ccRCC cell lines. ( D ) RT-PCR analysis of AK3 mRNA expression in ccRCC cell lines. ( E ) Immunofluorescence staining of AK3 protein in ccRCC cells. Labeling mitochondria with TOM20 antibody and AK3 protein with AK3 antibody. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Biomarker Discovery, Expressing, Western Blot, Control, Immunohistochemical staining, Microarray, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining, Labeling

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: Correlation Between AK3 Expression and Immune Infiltration in ccRCC. ( A ) Correlation analysis of AK3 expression with immune cell infiltration using the ssGSEA algorithm across 24 immune cell types. ( B ) Scatter plots depicting the correlation of AK3 expression with the enrichment scores of selected immune cell types: mast cells, T helper cells, Treg cells, and NK CD56bright cells. ( C ) Box plots comparing the expression levels of immune checkpoint genes (CD274, CTLA4, TIGIT, LAG3, PDCD1) between high AK3 (G1), low AK3 (G2), and normal groups. ( D ) Comparison of mutation frequencies of selected genes between high and low AK3 expression groups in the TCGA-KIRC cohort. ( E ) Correlation analysis of AK3 mRNA expression with drug sensitivity using the GSCA database. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Expressing, Comparison, Mutagenesis

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: AK3 Overexpression Inhibits Growth and Migration of ccRCC Cells. ( A ) Western blot analysis showing the successful overexpression of AK3 protein in 786O and OS ccRCC cell lines. ( B ) Relative RNA expression levels of AK3 in 786O and OS cells following transfection with AK3 overexpression plasmids, as determined by RT-PCR. ( C ) Cell growth curves for 786O and OS cells, illustrating a significant decrease in proliferation rates in the AK3 overexpression group compared to controls over a 5-day period. ( D ) Representative images and quantification of colony formation assays in 786O and OS cell lines. ( E ) Wound healing assay images and quantification demonstrating reduced migration of 786O and OS cells following AK3 overexpression. ( F ) Transwell migration assay results showing a significant decrease in the number of migrating 786O and OS cells upon AK3 overexpression. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Over Expression, Migration, Western Blot, RNA Expression, Transfection, Reverse Transcription Polymerase Chain Reaction, Wound Healing Assay, Transwell Migration Assay

Journal: International Journal of General Medicine

Article Title: AK3 as a Hypoxia-Angiogenesis–Related Prognostic Biomarker and Therapeutic Target in Clear Cell Renal Cell Carcinoma

doi: 10.2147/IJGM.S552108

Figure Lengend Snippet: AK3 Overexpression Inhibits the PI3K-AKT/GSK3β Signaling Pathway in ccRCC Cells. ( A ) Volcano plot depicting differential gene expression in 786O cells following transfection with AK3 overexpression plasmids. ( B ) KEGG pathway enrichment analysis highlighting downregulated pathways upon AK3 overexpression. ( C ) KEGG enrichment analysis of upregulated pathways in AK3-overexpressing cells. ( D ) Western blot analysis of key signaling proteins in the PI3K/AKT pathway. ( E ) In vivo analysis of tumor growth volume and weight in nude mice. ( F ) Immunohistochemical analysis of AK3 and Ki67 expression in tumor tissues. Data were shown as mean ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: Human-derived ccRCC cell lines (Caki-2, OS-RC-2, 786-O) and a normal renal tubular epithelial cell line (HK-2) were used, all obtained from Servicebio.

Techniques: Over Expression, Gene Expression, Transfection, Western Blot, In Vivo, Immunohistochemical staining, Expressing

Journal: Frontiers in Immunology

Article Title: Sialylation-immune-related lncRNA LINC01605 promotes tumor-infiltrating CD8 + T cell exhaustion and malignancy of clear cell renal cell carcinoma

doi: 10.3389/fimmu.2025.1744278

Figure Lengend Snippet: The highly expressed sialylation-immune-related lncRNA LINC01605 in ccRCC correlates with poor prognosis. (A) 66 overlapping lncRNAs associated with SRGs, immune cells, phenotypes, and prognosis were identified in a Venn diagram for further investigation. (B) C-index of prognostic models constructed by multiple machine learning methods. (C) Forest plot displaying univariate Cox regression analysis results for 12 SIRLs in OS, PFI, and DSS. The HR values along with their corresponding 95% confidence intervals are shown. (D) Kaplan-Meier plots for overall survival (OS), progression-free interval (PFI), and disease-specific survival (DSS) were generated using the median expression of LINC01605 as the cut-off. (E) Relative RNA expression of LINC01605 in 10 pairs of human ccRCC tumors (n = 10 per group). (F) FISH-IF detected LINC01605 expression in tumor and adjacent non-tumor tissues of ccRCC patients, with CA9 as the ccRCC cell marker. (G) Expression levels of LINC01605 in HK-2 and different ccRCC cell lines were detected (n = 3 per group). (H) GSEA revealed high LINC01605 expression to be functionally associated with enriched cell proliferation, stemness, migration, and invasion pathways. All bioinformatics analyses in Figure 1 were based on the gene expression data (TPM values) and clinical data of 528 ccRCC samples from the TCGA-KIRC cohort. Values are presented as mean ± SD. P -values were calculated by log-rank (Mantel-Cox) test (D) , two-tailed paired Student’s t-test (E) , one-way ANOVA followed Tukey’s multiple comparisons (G) , or non-parametric permutation test (H) . *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. n represents the number of biological replicates in (E, G) .

Article Snippet: The human embryonic kidney 293T cell line (293T), immortalized renal epithelial cell line (HK-2), and the human ccRCC cell lines (Caki-1, 769-P, Caki-2, 786-O, A-498 and RCCJF) were obtained from the American Type Culture Collection (ATCC).

Techniques: Construct, Generated, Expressing, RNA Expression, Marker, Migration, Gene Expression, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Sialylation-immune-related lncRNA LINC01605 promotes tumor-infiltrating CD8 + T cell exhaustion and malignancy of clear cell renal cell carcinoma

doi: 10.3389/fimmu.2025.1744278

Figure Lengend Snippet: LINC01605 is associated with the malignant progression of ccRCC cells. (A) LINC01605 was silenced in A498 and 786-O cell lines by two different shRNAs (n = 3 per group). (B, C) CCK8 and colony formation assays were performed in LINC01605 -knockdown and counterpart control groups (n = 3 per group). (D, E) Transwell migration/invasion and wound-healing assays showed that knockdown of LINC01605 impaired mobility and invasiveness of ccRCC cell lines (n = 3 per group). (F) EdU assay showed that cell proliferation and DNA synthesis decreased in LINC01605 -silenced ccRCC cell lines (n = 3 per group). (G) Knockdown of LINC01605 in A498 cells inhibited the growth of A498-derived xenograft in vivo (n = 5 per group). Tumor growth curves, tumor weights and statistical analysis are shown. Values are presented as mean ± SD. P -values were determined by one-way ANOVA followed Tukey’s multiple comparisons [ (A-G) one comparison per time point for (B , G) ]. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. n represents the number of biological replicates in (A-G) .

Article Snippet: The human embryonic kidney 293T cell line (293T), immortalized renal epithelial cell line (HK-2), and the human ccRCC cell lines (Caki-1, 769-P, Caki-2, 786-O, A-498 and RCCJF) were obtained from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Control, Migration, EdU Assay, DNA Synthesis, Derivative Assay, In Vivo, Comparison

Journal: Frontiers in Immunology

Article Title: Sialylation-immune-related lncRNA LINC01605 promotes tumor-infiltrating CD8 + T cell exhaustion and malignancy of clear cell renal cell carcinoma

doi: 10.3389/fimmu.2025.1744278

Figure Lengend Snippet: LINC01605 is associated with sialic acid levels and involved in JAK3/STAT3 signaling. (A, B) Uniform manifold approximation and projection (UMAP) plots of ccRCC samples and tumor-infiltrating CD8 + T cell subpopulation. (C) ccRCC samples with highly sialylated tumor cells had higher exhaustion scores and lower effector and cytotoxicity scores in the corresponding CD8 + T cell subpopulation. (D) Flow cytometry results indicated that LINC01605 silencing decreased sialic acid levels on the cell membrane of A498 and 786-O cell lines (n = 5 per group). (E) qRT-PCR and western blotting showed that LINC01605 expression was correlated with mRNA and protein expression of ST6GALNAC5 (n = 3 per group). (F) GSEA analysis in the TCGA-KIRC cohort (n=528) revealed that high LINC01605 expression was related to the IL6/JAK/STAT3 and MYC target pathways. (G) The mRNA expression of JAK3 and STAT3 was detected using qRT-PCR in LINC01605 -silenced ccRCC cell lines (n = 3 per group). (H) Western blotting was performed to detect the expression of JAK3, STAT3, phosphorylated STAT3, and downstream genes of the pathway after LINC01605 knockdown in ccRCC cell lines. (I) Binding sites of STAT3 at the ST6GALNAC5 promoter were predicted by JASPAR. (J) ChIP assays demonstrated that STAT3 bound to the ST6GALNAC5 promoter in A498 and 786-O cells (n = 3 per group). Values are presented as mean ± SD. P -values were calculated by two-tailed unpaired Student’s t-test (C, J) , one-way ANOVA followed Tukey’s multiple comparisons (D, E, G) , or non-parametric permutation test (F) . *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. n represents the number of samples in (F) and the number of biological replicates in (D-E, G, J) .

Article Snippet: The human embryonic kidney 293T cell line (293T), immortalized renal epithelial cell line (HK-2), and the human ccRCC cell lines (Caki-1, 769-P, Caki-2, 786-O, A-498 and RCCJF) were obtained from the American Type Culture Collection (ATCC).

Techniques: Flow Cytometry, Membrane, Quantitative RT-PCR, Western Blot, Expressing, Knockdown, Binding Assay, Two Tailed Test