murine rcc cell line renca  (Procell Inc)

Journal: Journal of Cancer

Article Title: Dual-Inhibition of mTOR and Bcl-2 Enhances the Anti-tumor Effect of Everolimus against Renal Cell Carcinoma In Vitro and In Vivo

doi: 10.7150/jca.29192

Figure Lengend Snippet: Effect of everolimus and/or ABT-737 on RenCa and RenCa res cells. ( A ) Everolimus response of parental RenCa cells was characterized by investigating the effect of increasing everolimus concentration on cell viability at 24, 48, and 72 hours. ( B ) RenCa cells were subjected to increasing concentrations of ABT-737 at indicated time points. (C) RenCa res cells were subjected to increasing doses of everolimus for 24 to 72 hours to test acquired drug resistance by performing cell viability assay. ( D ) The response of RenCa res cells to ABT-737 monotherapy was investigated by cell viability assay upon the treatment of cells with various ABT-737 concentrations for indicated time points. Cell viability of untreated parental and resistant control cells was considered as 100% for 24 hours and graphs show the mean ± S.D of three independent experiments, each performed in triplicate. *, P ≤ 0.05 ; **, P ≤ 0.01 ; ***, P ≤ 0.001 ; ****, P ≤ 0.0001 . ( E ) The effect of the acquired everolimus resistance on mTOR pathway was determined in cell lysates of RenCa and RenCa res cells by investigating the protein levels of p-mTOR (Ser2448), mTOR, p-p70S6K (Thr389), p70S6K, p-S6 (Ser235/236), p-S6 (Ser240/244), S6, p-4E-BP1 (Ser65), 4E-BP1. ( F ) Western blot analysis of Bcl-2, Bcl-xL, and Mcl-1 was performed with proteins isolated from RenCa and RenCa res cells. β-Actin was used as loading control. ( G ) RenCa res cells exposed to 20 µM ABT-737 were used for Annexin-V/PI staining to determine the effect of ABT-737 monotherapy for 72 hours. The graph shows the percentage of Annexin V-positive cells. Each data value represents the mean ± S.D. of three independent experiments, each performed in triplicate. *, P ≤ 0.05.

Article Snippet: The human RCC cell lines A-498, Caki-2, Caki-1, ACHN, HEK-293 and mouse murine RCC cell line RenCa were purchased from American Type Culture Collections (ATCC).

Techniques: Concentration Assay, Viability Assay, Control, Western Blot, Isolation, Staining

Journal: Journal of Cancer

Article Title: Dual-Inhibition of mTOR and Bcl-2 Enhances the Anti-tumor Effect of Everolimus against Renal Cell Carcinoma In Vitro and In Vivo

doi: 10.7150/jca.29192

Figure Lengend Snippet: Anti-tumor effect of everolimus and ABT-737 combination on RenCa res xenograft mice model. BALB/c mice were randomized into four groups (n = 8) and administered every other day with the vehicle control, 2 mg/kg everolimus, 75 mg/kg ABT-737 or the combination of single drug doses. ( A ) Representative pictures of tumors isolated from each group after 21 days of treatment were shown. ( B ) Average weight of tumors from each cohort subjected to everolimus and/or ABT-737 treatment or control was shown. Scatter blot represents the mean ± S.D. The significant analysis of tumor weights of control groups compared to treatment groups was performed by two-tailed Student's t-test. *, P ≤ 0.05 ; ****, P ≤ 0.0001 . ( C ) Images are the representatives for hematoxylin and eosin staining of tumor tissue sections. ( D ) The graph shows the change of mice body weight at the days of injection. Each data point in the graph represents the mean ± S.D. of body weights of eight mice in each group.

Article Snippet: The human RCC cell lines A-498, Caki-2, Caki-1, ACHN, HEK-293 and mouse murine RCC cell line RenCa were purchased from American Type Culture Collections (ATCC).

Techniques: Control, Isolation, Two Tailed Test, Staining, Injection