RNA interference targeting protein kinase C iota inhibits cell proliferation of cervical carcinoma cell line HeLa
Lihua Gao, Ya-ping Wen, Qizhi Luo, Ping Yu, Meiyu Yan, Ming Li
Abstract
Lihua Gao, Ya-ping Wen, Qizhi Luo, Ping Yu, Meiyu Yan, Ming Li
Abstract
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. Key words: Cervical carcinoma ; RNA interference ; Proliferation ; Apoptosis
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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. Key words: Cervical carcinoma ; RNA interference ; Proliferation ; Apoptosis
Key concepts: HeLa, Transfection, Small hairpin RNA, Molecular biology, Apoptosis, Cell growth, Cell culture, Cell cycle