primary renal cortical epithelial cells hrce  (ATCC)

Journal: BMC Cancer

Article Title: Preclinical efficacy of dual mTORC1/2 inhibitor AZD8055 in renal cell carcinoma harboring a TFE3 gene fusion

doi: 10.1186/s12885-019-6096-0

Figure Lengend Snippet: Cell viability, cytotoxicity and cell cycle progression in TfRCC cell lines treated with mTOR inhibitors. a , b Cell viability, as measured by MTT assay for TfRCC cell lines and the benign renal epithelial cell line HRCE after 72 h of treatment with up to 1000 nM concentrations of the dual mTORC1/2 inhibitor, AZD8055 ( a ), or selective mTORC1 inhibitor, sirolimus ( b ). Viability in TfRCC cells was suppressed by approximately 80–90% with AZD8055 and 30–50% with sirolimus relative to the untreated (0 nM drug) condition. Both drugs inhibited growth to a greater degree in TfRCC cells than in benign renal cells. c , d Cell cytotoxicity, as measured by LDH release by UOK120 and UOK146 TfRCC cell lines after 48 h of treatment with 1 μM of AZD8055 ( c ) or sirolimus ( d ). Only slight cytotoxicity in UOK120 cells and no cytotoxicity in UOK146 cells was observed after AZD8055 treatment, while sirolimus treatment had no cytotoxic effect. Multi protein inhibitor LY294002 [100 μM] was used as a positive control. e , f Relative fraction of cells in S-phase of the cell cycle, as measured by BrdU incorporation in UOK120 ( e ) and UOK146 ( f ) cell lines treated for 24 h with low (50 nM) and high (500 nM) concentrations of AZD8055 or sirolimus. Dose-dependent reductions in S-phase in both cell lines with either drug mirror the magnitude of reductions observed in cell viability ( a , b ), supporting a predominantly cytostatic mechanism of growth inhibition for both drugs. * p < 0.05; ** p < 0.01; *** p < 0.001; NS = non-significant

Article Snippet: RCC4 was obtained from ECACC General Cell Collection (Salisbury, UK; Cat Nr. 03112702) and the human renal cortical epithelial (HRCE) cell line was obtained from ATCC (Manassas, VA; Cat Nr. PCS-400-011).

Techniques: MTT Assay, Positive Control, BrdU Incorporation Assay, Inhibition

Journal: Molecular and Cellular Biology

Article Title: RNF20 Suppresses Tumorigenesis by Inhibiting the SREBP1c-PTTG1 Axis in Kidney Cancer

doi: 10.1128/MCB.00265-17

Figure Lengend Snippet: RNF20 is downregulated in ccRCC. (A) qRT-PCR analysis of RNF20 mRNA expression in patient-matched normal kidney (n = 9) and ccRCC tumor (n = 9) samples. RNF20 mRNA levels were normalized to those in matched normal kidney samples. (B) Normalized RNA-seq reads of RNF20 in normal kidney (n = 72) and ccRCC tumor (n = 533) samples. RNA-seq data were obtained from TCGA. (C) RNF20 expression in ccRCC tumors was analyzed according to tumor stages. Significance versus normal kidney samples: ##, P < 0.01; ###, P < 0.001. (D) Representative ccRCC tissue microarray used for IHC with RNF20 antibody. (E) IHC staining of patient-matched adjacent normal kidney and ccRCC tumor tissues. Representative tissue sections stained for RNF20 are shown. Bar, 100 μm. (F) RNF20 protein expression levels in normal kidney cell lines, such as HRCE and HEK293, and ccRCC cell lines, including ACHN, A498, and Caki-2, were determined by Western blotting. (G) RNF20 mRNA expression in normal kidney and ccRCC cell lines was determined by qRT-PCR. Significance versus HRCE: ##, P < 0.01.

Article Snippet: ACHN, A498, HEK293, Caki-2, and human primary renal cortical epithelial (HRCE) cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Quantitative RT-PCR, Expressing, RNA Sequencing, Microarray, Immunohistochemistry, Staining, Western Blot

Journal: Molecular and Cellular Biology

Article Title: RNF20 Suppresses Tumorigenesis by Inhibiting the SREBP1c-PTTG1 Axis in Kidney Cancer

doi: 10.1128/MCB.00265-17

Figure Lengend Snippet: RNF20 suppresses cell growth in ccRCC but not in normal kidney cell lines. (A) ACHN and A498 ccRCC cells were infected with adenovirus expressing GFP alone (−) or Myc-RNF20(+). After infection for 24 h, total cell lysates were subjected to Western blotting. (B) ACHN and A498 ccRCC cells were infected with adenovirus expressing GFP alone (Mock) or RNF20, and proliferation was monitored using the Cell Counting Kit-8 (CCK-8) assay. (C) ACHN and A498 ccRCC cells were transfected with siControl or siRNF20, and RNF20 expression was determined by Western blotting. (D) ACHN and A498 ccRCC cells were transfected with siControl or siRNF20, and relative growth rates were determined using the CCK-8 assay. (E) HRCE and HEK293 normal kidney cells were infected with adenovirus expressing GFP alone (−) or Myc-RNF20(+), and cell lysates were examined using Western blotting. (F) HRCE and HEK293 cells were infected with adenoviral RNF20, and cell proliferation was monitored using the CCK-8 assay. (G) HRCE and HEK293 cells were transfected with siControl or siRNF20, and cell lysates were determined by Western blotting. (H) HRCE and HEK293 cells were transfected with siRNF20, and cell proliferation rates were monitored using the CCK-8 assay. Cell proliferation data are presented as the means ± SD from five individual samples. *, P < 0.05; **, P < 0.01; n.s., not significant.

Article Snippet: ACHN, A498, HEK293, Caki-2, and human primary renal cortical epithelial (HRCE) cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Infection, Expressing, Western Blot, Cell Counting, CCK-8 Assay, Transfection