CAJ | 학술논문

目的观察小分子干扰RNA(siRNA)沉默蛋白激酶Cι (PKCι)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PKCι基因的siRNA转染HeLa细胞,噻唑蓝(MTT)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PKCι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±4.54)%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)%和空白对照组(2.75±0.24)%.瞬时转染PKCι siRNA 72h后,G1期细胞增多,G2+S期细胞减少.结论沉默PKCι可抑制宫颈癌细胞增殖,促进细胞凋亡.
목적 관찰소분자간우RNA(siRNA)침묵단백격매Cι (PKCι)표체대HeLa증식여조망적영향.방법 실험분위전염조、음성대조화공백대조조.장PKCι기인적siRNA전염HeLa세포,새서람(MTT)비색법검측3조세포제1～5천적흡광도;배양세포우제21천계수세포극륭수;류식세포의검측침묵PKCι기인72h적세포조망급세포주기분포.결과 전염조세포증식솔저우량개대조조.전염조적평판집락형성솔(64.00±4.54)%저우음성(87.00±1.22)%화공백조(82.00±2.10)%(P<0.05).전염조세포조망솔(9.54±0.55)%명현고우음성대조조(1.31±0.10)%화공백대조조(2.75±0.24)%.순시전염PKCι siRNA 72h후,G1기세포증다,G2+S기세포감소.결론 침묵PKCι가억제궁경암세포증식,촉진세포조망.
Objective To explore the effect of RNA interference (RNAi) targeting protein kinase C iota (PKCι) on proliferation and apoptosis of cervical carcinoma cell line HeLa. Methods Stable cell lines which were transfected with PKCι 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 1st-5th day. The cells were cultured in 6-well plates and stained by crystal violent at 21st 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 PKCι shRNA showed markedly decrease of proliferation assayed by MTT and plate colony formation. The apoptosis rate of cells transiently transfected with PKCι shRNA (9.54±0.55)% was higher than that of cells transiently transfected with psi-mock (1.31±0.10)% or HeLa blank control cells (2.75±0.24)% (P<0.05). The cells transiently transfected by PKCι shRNA were arrested in G0/G1 phase. Conclusion PKCι is essential for maintaining of HeLa cell proliferation. Silencing PKCι can inhibit the growth of HeLa cells and induce apoptosis.