小分子干扰RNA降低蛋白激酶Cl表达抑制HeLa细胞增殖  

RNA interference targeting protein kinase C iota inhibits cell proliferation of cervical carcinoma cell line HeLa

作　　者：高丽华[1] 文亚平[1] 罗奇志[1] 余平[1] 燕美玉[1] 黎明[1] 

机构地区：[1]中南大学湘雅基础医学院免疫学教研室,长沙410078

出　　处：《中华实验外科杂志》2011年第7期1175-1178,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察小分子干扰RNA（siRNA）沉默蛋白激酶Cl（PKCL）表达对HeLa增殖与凋亡的影响。方法实验分为转染组、阴性对照和空白对照组。将PKCL基因的siRNA转染HeLa细胞，噻唑蓝（MTF）比色法检测3组细胞第1～5天的吸光度；培养细胞于第21天计数细胞克隆数；流式细胞仪检测沉默PKCl基因72h的细胞凋亡及细胞周期分布。结果转染组细胞增殖率低于两个对照组。转染组的平板集落形成率（64．00±d．54）％低于阴性（87．00±1．22）％和空白组（82．00±2．10）％（P〈0．05）。转染组细胞凋亡率（9．54±0．55）％明显高于阴性对照组（1．31±0．10）％和空门对照组（2．75±0．24）％。瞬时转染PKCL siRNA 72h后，G1期细胞增多，G2＋S期细胞减少。结论沉默PKCl可抑制宫颈癌细胞增殖，促进细胞凋亡。Objective To explore the effect of RNA interference （RNAi） targeting protein kinase C iota （PKCl） on proliferation and apoptosis of cervical carcinoma cell line HeLa. Methods Stable cell lines which were transfected with PKCl shRNA or vector-mock plasmids were established. The cell lines, psi-PKCiota-HeLa, psi-mock-HeLa and HeLa, were used to detect methyl thiazol tetrazolium （MTT） absorbance at 1 st-5th day. The cells were cultured in 6-well plates and stained by crystal violent at 21 st day, and the stained cells colonies were counted. Flow cytometry was used to detect apoptosis and cell cycle distribution after HeLa cells were transiently transfected with PKC iota shRNA at 72nd h. Results Compared with the psi-mock and blank controls, the transfected cells stably expressing PKCl shRNA showed markedly decrease of proliferation assayed by MTT and plate colony formation. The apoptosis rate of cells transiently transfected with PKCL shRNA （9. 54 ± 0. 55 ） % was higher than that of cells transiently transfected with psi-mock （ 1.31 ±0. 10） % or HeLa blank control calls （2. 75 ±0. 24） % （ P 〈 0. 05 ）. The cells transiently transfected by PKCl shRNA were arrested in G0/G1 phase. Conclusion PKCl is essential for maintaining of HeLa cell proliferation. Silencing PKCl can inhibit the growth of HeLa cells and induce apoptosis.

关 键 词：宫颈癌 RNA干扰 增殖 脱噬作用 

分 类 号：R730.2[医药卫生—肿瘤]