Effect of Dicer knockdown on proliferation and migration of clear cell renal cell carcinoma cell line 769-P

Abstract

Objective To observe the effect of Dicer knockdown on proliferation and migration of clear cell renal cell carcinoma cell line 769-P.Methods Liposome-mediated method was used to transfect siRNA sequence targeting Dicer and mock sequence as control into 769-P cells.Dicer expression levels after transfection were detected by using real-time polymerase chain reaction (Real-time PCR) and Western blotting.3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and Transwell assays were used to examine the changes in proliferation and migration of 769-P cells.Results Compared to untransfected group and Mock group,the mRNA levels in siRNA group were significantly reduced at 24,48 and 72 h after transfection (0.001 562 ± 0.000 132,0.000 492 ±0.000 039 and 0.000 752 ±0.000 114,respectively,P ＜0.01),with the effect most obvious at 48 h after transfection.The protein levels in siRNA group were also significantly reduced (1.062 ± 0.207,P ＜0.01).MTS assay revealed the absorbance (A) value in siRNA group was significantly increased at 24,48,72 and 96 h after transfection (0.451 ±0.016,0.544 ±0.017,0.612 ±0.016 and 0.651 ±0.015,respectively,P ＜ 0.01),and Transwell assay showed the number of migratory cells in siRNA group was significantly increased at 48 h after transfection (93.100 ± 8.999,P ＜ 0.01).Conclusion Knockdown of Dicer significantly enhanced proliferation and migration of 769-P cells. Key words: Dicer; Clear cell renal cell carcinoma; Proliferation; Migration