Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: Inhibitory effects of miR‐33a on cell proliferation and cell cycle in renal cell cancer (RCC) cell lines. (a) Expression of miR‐33a in Caki‐1 and 786‐O cells after transfection with miR‐33a mimics and miR‐33a inhibitor or NC. NC represents negative control of miRNA. Results were expressed as of three independent experiments, with at least three replicates in each independent experiment, ** p < 0.01 versus NC. (b) CCK‐8 assays indicated that the effects of miR‐33a on the growth of RCC cell lines. Results were expressed as of three independent experiments, with at least three replicates in each independent experiment, * p < 0.05 versus NC. (c, d) The results of flow cytometry showed that upregulation of miR‐33a significantly increased the percentages of cells in the G0/G1 phase, which showed that the upregulation of miR‐33a could suppress RCC cells proliferation. * p < 0.05 versus Control

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Expressing, Transfection, Negative Control, CCK-8 Assay, Flow Cytometry, Control

Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: miR‐33a directly targets Mouse double minute 4 (MDM4). (a) The predicted miR‐33a binding site within MDM4 3′ UTR and miR‐33a mutated version by site mutagenesis; (b) Caki‐1 cells were transfected with reporter constructs containing either wild type (WT) MDM4 , or MDM 4 3′ UTR with mutation (MUT), along with miR‐33a mimics, or negative control, respectively. Relative luciferase activity was measured. (c) qPCR analysis of MDM4 expression in renal cell cancer (RCC) Caki‐1 cells after overexpression or knockdown of miR‐33a . (d, e) Western blotting analysis of MDM4 protein expression in RCC Caki‐1 cells after overexpression or knockdown of miR‐33a . (f) Western blot analysis revealed that transfection of MDM4 siRNA into Caki‐1 cells resulted in decreased MDM4 expression compared to the cells transfected with scrambled siRNA. These effects of siRNA were attenuated by anti‐ miR‐33a inhibitor transfection. NC represents normal control, * p < 0.05, ** p < 0.01 versus NC

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Binding Assay, Mutagenesis, Transfection, Construct, Negative Control, Luciferase, Activity Assay, Expressing, Over Expression, Knockdown, Western Blot, Control

Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: The effect of mouse double minute 4 ( MDM4 ) on cell growth in renal cell cancer (RCC). (a, b, c) RT‐qPCR and western blot analysis of MDM4 expression in RCC cells. (d) Kaplan–Meier analysis of overall survival in 30 RCC patients with low median ( n = 15) and high median ( n = 15) expression levels of MDM4 . (e) CCK‐8 assays indicated that the effects of MDM4 on growth of RCC cell lines. Results were expressed as ± x - s of three independent experiments, with at least three replicates in each independent experiment, * p < 0.05 versus NC. (f) Western blot analysis of p53 expression after miR‐33a mimics and ‐inhibitor transfection in RCC cells

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Transfection