The correlation of E2F3, miR-17-5p and miR-20a in transitional cell carcinoma of bladder

Abstract

Objective To explore the correlation and role of E2F3 gene,miR-17-5p and miR-20a in the cell lines of transitional cell carcinoma of bladder. Methods The plasmids of pcDNA3.1-HA-E2F3 and pAAV-siRNA-E2F3 were used to overexpress and knockdown E2F3.The mimics of miR-17-5p,miR-20a and their anti-miRNA oligonucleotides were used to overexpress and screen miR-17-5p and miR-20a.The expression levels of E2F3 gene,miR-17-5p and miR-20a were detected by quantitative real-time PCR,and E2F3 protein were detected by Western blot. Results When E2F3 was overexpressed,the 2- △△Ct of miR-17-5p and miR-20a were 2.26 ± 0.30 and 4.04 ± 0.51,it was statistically significant to compared with control (P ＜ 0.05) ; when E2F3 was knockdown,the 2 △△Ct of miR-17-5p and miR-20a were 0.49 ± 0.02and 0.65 ± 0.04 (P ＜ 0.05) ; when miR-17-5p and miR-20a were overexpressed simultaneously,the level of E2F3 mRNA was significantly decreased,the average E2F3 protein gray scale was 55.31 ± 7.89,the control was 103.67 ± 13.61 (P ＜ 0.05 ) ; when miR-17-5p and miR-20a were knockdown simultaneously,the E2F3 mRNA was significantly increased,the E2F3 protein gray scale was 295.68 ± 19.25,the control was 103.67 ± 13.61 ( P ＜ 0.05 ). Conclusions miR-17-5p and miR-20a could be up-regulated by E2F3 gene,and the E2F3 gene could be down-regulated by miR-17-5p and miR-20a.The regulatory feedback loop of E2F3 gene,miR-17-5p and miR-20a exists in transitional cell carcinoma of bladder. The loop maybe plays a key role in the development of bladder cancer. Key words: E2F3 gene; Micro RNA; Bladder neoplasms; Gene regulation