Effect of Dicer knockdown on proliferation and migration of clear cell renal cell carcinoma cell line 769-P
Fan Yang, Yu Gao, Qingbo Huang, Qing Ai, Xin Ma, Xu Zhang
Abstract
Fan Yang, Yu Gao, Qingbo Huang, Qing Ai, Xin Ma, Xu Zhang
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
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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
Key concepts: Dicer, Transfection, Gene knockdown, Molecular biology, Small interfering RNA, Cell growth, Cell culture, Chemistry