Effect of E2F1 Knockdown on proliferation and invasion of clear cell renal cell carcinoma cell line Caki-2

Abstract

Objective To study the effect of E2F1 knockdown on proliferation,migration and invasion of clear cell renal cell carcinoma cell line Caki-2.Methods Small interfering RNA (siRNA)-E2F1and negative control sequences were transfected into Caki-2 cells by Lipo2000.The E2F1 expression levels were detected by using real-time quantitative polymerase chain reaction (PCR) and Western blotting respectively after transfection.MTS and Transwell assays were used to detect variations in cell proliferation,migration and invasion.Results Compared to untransfected group (130.100 ± 6.301) × 10-5 and negative control group (134.600 ± 7.163) × 10-5,the mRNA levels in siRNA-E2F1 group (37.770 ± 3.782) × 10-5were significantly reduced (P ＜0.01).The protein levels were consistent with mRNA levels.MTS assay demonstrated the absorbance in siRNA-E2F1 group was significantly decreased at the time points of 24,48,72 and 96 h after transfection (P ＜ 0.05),and Transwell assay revealed that the number of migrating cells in siRNA-E2F1 group (38.330 ±4.041) was significantly reduced at 48 h after transfection as compared with untransfected group (83.000 ± 8.888) and negative control group (87.330 ± 6.429) (P ＜0.01),and the number of invasive cells in siRNA-E2F1 group (78.330 ±7.572) was strongly reduced at 48 h after transfection as compared with untransfected group (143.30 ± 10.066) and negative control group (147.000 ± 14.000) (P ＜0.01).Conclusion Knockdown of E2F1 significantly decreased proliferation,migration and invasion of Caki-2 cells. Key words: Clear cell renal cell carcinoma; E2F1 gene ; Proliferation; Migration; Invasion