Effect of miR-103b on the growth of renal cell carcinoma by activating the expression of P21 protein

Abstract

Objective To investigate the effect of miR-103b on the expression of P21 protein in renal cell carcinoma cell line 769-P and ACHN cells, and its effect on the growth of renal cell carcinoma. Methods Renal cancer cells were divided into two groups according to the transfected RNA, miR-103b (experimental group) and dsControl (control group), respectively. Real-time PCR and Western blotting were used to detect the expression of P21, cell cycle-dependent kinase 6, Cyclin D1 mRNA and protein expression. Flow cytometry was used to detect the cell cycle distribution. MTT assay was used to detect cell viability and colony formation assay was used to detect cell proliferation. Measurement data were represented as ±s. Comparison between groups was analyed using t test. Results Real-time PCR results showed that the relative expression levels of P21, cell cycle-dependent kinase 6 and Cyclin D1 mRNA in 769-P and ACHN which belong to control group cells were 1.00±0.10 and 1.02±0.27, 1.00±0.08 and 1.01±0.17, 1.01±0.19 and 1.00±0.02. The experimental group was 2.36±0.51 and 2.03±0.49, 0.33±0.20 and 0.58±0.22, 0.48±0.11 and 0.60±0.23, respectively, and the difference was statistically significant (P<0.05). Western blotting results were consistent with Real-time PCR results. Flow cytometry results showed that compared with the control group, the proportion of cells located in G0/G1 phase in the experimental group increased (P<0.05), suggesting that the cells were arrested in G0/G1 phase. MTT assay showed that the viability of 769-P and ACHN cells in the experimental group was significantly lower than that in the control group. Colony formation experiments showed that the number of colony formation in the experimental group was significantly less, suggesting that the cell proliferation capacity decreased. Conclusion miR-103b can inhibit the growth of renal cell carcinoma cells by activating the expression of P21 protein and blocking the progression of the renal cell cycle, which provides a theoretical basis for the molecular targeted therapy of renal cell carcinoma. Key words: MicroRNA; Renal tumor; Cell proliferation; RNA activation