tubular epithelial cell line hk2  (Procell Inc)

Journal: International Journal of Biological Sciences

Article Title: Suppression of ASNS expression by VHL-mediated ubiquitination hinders the progression of renal cell carcinoma through enhancing JUP expression and inhibiting PI3K-AKT and MAPK pathways

doi: 10.7150/ijbs.129332

Figure Lengend Snippet: L-Asparagine enhanced the growth and metastasis of RCC in vitro and in vivo . Cell plate cloning experiments assessing HK2, Caki-1 and 786-O cell lines treated with varying concentrations of L-Asparagine (L-Asn). (B) Cell plate cloning experiments evaluating HK2, Caki-1, and 786-O cell lines treated with L-ASNase (0.5 U/mL). (C and D) Experiments measuring cell migration and invasion in HK2, Caki-1, and 786-O cell lines treated with either L-Asn (0.5mM) or L-ASNase (0.5 U/mL). (E) In vivo imaging of mouse orthotopic renal tumors fed with either L-Asn-rich (3g L-Asn/kg feed) or deficient diets (Each group consists of 5 mice). (F) In vitro imaging of mouse lungs subjected to L-Asn-rich (3g/kg) or -deficient diets (Each group consists of 5 mice). Lung metastasis rate: CON: 5/5; L-Asn -: 4/5; L-Asn +: 5/5. (G) ELISA assay quantifying the levels of L-Asparagine synthase (ASNS) and L-ASNase in mouse kidney tumors. (H) ELISA assay quantifying the levels of ASNS and L-ASNase in plasma. (I) The relationship between the expression of ASNS in the plasma of these 63 VHL -mutant RCC patients and the disease-free survival (DFS) of patients. (J) The expression of ASNS in these 19 pairs of VHL -mutant RCC and their AN tissues was detected through immunohistochemical staining. (K) Immunohistochemical staining to detect ASNS protein in ccRCC tissues. (L) Western blotting analysis of ASNS protein in RCC tissues. (M) Western blotting analysis of ASNS protein in various RCC cell lines (A498, OSRC2, Caki-1, 769-P, 786-O, and RCC4).

Article Snippet: Six RCC cell lines A498, OSRC2, Caki-1 ( VHL wild-type, metastatic RCC cell line), 769-P, 786-O ( VHL mutant), RCC4 ( VHL mutant) and two control cell lines HK2 (Epithelial cells of human renal cortex proximal tubules), HEK-293 (Human embryonic kidney cells) were purchased from the ATCC (American Type Culture Collection) cell bank.

Techniques: In Vitro, In Vivo, Cloning, Migration, In Vivo Imaging, Imaging, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Expressing, Mutagenesis, Immunohistochemical staining, Staining, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Suppression of ASNS expression by VHL-mediated ubiquitination hinders the progression of renal cell carcinoma through enhancing JUP expression and inhibiting PI3K-AKT and MAPK pathways

doi: 10.7150/ijbs.129332

Figure Lengend Snippet: VHL interacted with ASNS protein and modulated its expression. (A) Western blotting analysis showing changes in ASNS protein expression in HK2, 786-O, RCC4, and Caki-1 cell lines after treatment with different concentrations of L-Asn. (B) Comparison of ASNS protein expression in HK2, 786-O, RCC4, and Caki-1 cell lines post L-Asn treatment. (C) ELISA analysis of L-Asn expression levels in 786-O and RCC4 cell lines following VHL overexpression. (D and E) Western blotting analysis of ASNS expression changes in 786-O and RCC4 cell lines after VHL overexpression. (F) Western blotting analysis of ASNS expression changes in HEK-293 and Caki-1 cell lines after VHL knockdown. (G) Western blotting analysis of ASNS expression levels in Caki-1 cells after VHL knockout. (H) Confirmation of the regulatory effects of L-Asn and VHL on ASNS expression in 786-O, RCC4, and Caki-1 cell lines.

Article Snippet: Six RCC cell lines A498, OSRC2, Caki-1 ( VHL wild-type, metastatic RCC cell line), 769-P, 786-O ( VHL mutant), RCC4 ( VHL mutant) and two control cell lines HK2 (Epithelial cells of human renal cortex proximal tubules), HEK-293 (Human embryonic kidney cells) were purchased from the ATCC (American Type Culture Collection) cell bank.

Techniques: Expressing, Western Blot, Comparison, Enzyme-linked Immunosorbent Assay, Over Expression, Knockdown, Knock-Out

Journal: International Journal of Biological Sciences

Article Title: Suppression of ASNS expression by VHL-mediated ubiquitination hinders the progression of renal cell carcinoma through enhancing JUP expression and inhibiting PI3K-AKT and MAPK pathways

doi: 10.7150/ijbs.129332

Figure Lengend Snippet: Silencing ASNS and using an ASNS inhibitor significantly reduced RCC growth and metastasis in vitro and in vivo . The impact of ASNS overexpression and knockdown on the growth of HK2, Caki-1 and 786-O cell lines was evaluated using a colony formation assay. (B and C) Transwell migration and invasion assays were performed to assess the effects of ASNS overexpression and knockdown on the migration and invasion capabilities of HK2, Caki-1, and 786-O cell lines. (D) The influence of ASNS overexpression and knockdown on the growth and spontaneous lung metastasis of orthotopic renal tumors in mice was analyzed. (E) The effects of ASNS overexpression and knockdown on EdU, TUNEL, and Cleaved-Caspase 3 expression in mouse renal tumors were examined. (F) The impact of ASNS overexpression and knockdown on E-cadherin and N-cadherin expression in mouse lung metastases was assessed. (G) The effect of the ASNS inhibitor (Bisabosqual A) on the growth of Caki-1 and 786-O cell lines was evaluated through cell plate cloning experiments. (H and I) The influence of ASNS inhibitor on the migration and invasion of Caki-1 and 786-O cell lines was analyzed through migration and invasion assays. (J) The effect of ASNS inhibitor on the growth and spontaneous lung metastasis of mouse renal tumors was assessed. (K) The effects of ASNS inhibitor on EdU, TUNEL, and Cleaved-Caspase 3 expression in mouse renal tumors were examined. (L) The impact of ASNS inhibitor on E-cadherin and N-cadherin expression in mouse lung metastases was analyzed.

Article Snippet: Six RCC cell lines A498, OSRC2, Caki-1 ( VHL wild-type, metastatic RCC cell line), 769-P, 786-O ( VHL mutant), RCC4 ( VHL mutant) and two control cell lines HK2 (Epithelial cells of human renal cortex proximal tubules), HEK-293 (Human embryonic kidney cells) were purchased from the ATCC (American Type Culture Collection) cell bank.

Techniques: In Vitro, In Vivo, Over Expression, Knockdown, Colony Assay, Migration, TUNEL Assay, Expressing, Cloning

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Upregulated NNMT sustains angiogenesis in clear cell renal cell carcinoma through activation of the SDF-1/CXCR4–ERK–CCL20 axis

doi: 10.1007/s13402-025-01115-z

Figure Lengend Snippet: Increased expression of NNMT is observed in ccRCC and predicts an unfavorable prognosis for patients. ( A ) Heatmap of DEGs in normal samples and RCC samples. ( B ) Heatmap of methylation levels of methylation-driven genes (MDGs). ( C ) Venn diagram revealed that identification of 11 DEMDGs in RCC. ( D ) The expression levels and relative methylation levels of 11DEMDGs. ( E ) Kaplan-Meier survival curve showed the correlation of NNMT expression with survival rate of ccRCC in TCGA database. ( F ) The relative methylation level of NNMT. ( G ) Analysis of NNMT mRNA expression in TCGA-KIRC and GEO ( GSE66272 , GSE105261 ) datasets. ( H ) Scatterplot showing the fold change in NNMT mRNA levels in ccRCC and corresponding paracarcinoma specimens ( n = 20). ( I ) Western blot of NNMT expression in 9 pairs of ccRCC and corresponding paracarcinoma specimens. ( J ) IHC of NNMT expression in ccRCC and corresponding paracarcinoma specimens. The data are presented as a representative image. Scale bar, 50 μm. K . Correlation between overall survival (OS) and the relative expression level of NNMT in patients with ccRCC. L . Western blot of NNMT expression in the HK2 and RCC cell lines. Data were analyzed using Student’s t-test or Mantel-Cox. Significant results were presented as * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The ccRCC cell lines 769-P, 786-O, OS-RC2, and ACHN, along with the human immortalized proximal tubule epithelial cell line HK2, human umbilical vein endothelial cells (HUVEC), and HEK293T cells, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Methylation, Western Blot