抑制Dicer基因对肾透明细胞癌细胞株769-P细胞增殖和迁移的影响  

Effect of Dicer knockdown on proliferation and migration of clear cell renal cell carcinoma cell line769-P

作　　者：范阳[1] 高宇[1] 黄庆波[1] 艾青[1] 马鑫[1] 张旭[1] 

机构地区：[1]解放军总医院泌尿外科,北京100853

出　　处：《中华实验外科杂志》2013年第2期336-338,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30972982）

摘　　要：目的观察抑制Dicer基因后对肾透明细胞癌细胞株769一P细胞增殖和迁移的影响。方法选取肾透明细胞癌细胞株769．P，应用脂质体介导方法转染DicersiRNA序列和对照DicerMock序列细胞，采用实时定量聚合酶链反应（Real—timePCR）和Westernblot方法检测转染后Dicer的表达，并通过MTS和Transwell方法检测细胞增殖和细胞迁移能力的变化。结果与未转染组和转染DicerMock序列组比较，转染DicersiRNA序列组的DicermRNA水平在转染后24、48、72h明显降低（分别为0．001562±0．000132、0．000492±0．000039、0．000752±0．0001lg，P〈0．01），且在转染后48h最明显，转染DicersiRNA序列组的蛋白水平在转染后48h也明显降低（1．062±0．207，P〈0．01）。转染DicersiRNA序列组的MTS吸光度（A）值在转染后24、48、72、96h明显增高（分别为0．451-4-0．016、0．544-4-0．017、0．612±0．016、0．651-4-0．015，P〈0．01），Transwell穿膜细胞数在转染后48h也明显增多（93．100±8．999，P〈0．01）。结论抑制Dicer基因在肾透明细胞癌细胞株769一P中表达后，其细胞增殖能力和细胞迁移能力明显增强。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 af- ter 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 sig- nificantly 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.

关 键 词：DICER 肾透明细胞癌 增殖 迁移 

分 类 号：R737[医药卫生—肿瘤]