Mechanism and influence of silencing TGM1 gene by siRNA on cell cycle distribution of keratinocytes].

To explore the effects of cell cycle distribution and expression of cell cycle related proteins before and after transglutaminase 1 (TGM1) gene silencing by small interfering RNA (siRNA) in immortalized human keratinocytes (HaCaT cells). Before and after RNA interference (RNAi) silencing TGM1 gene expression in HaCaT cells, the changes of cell cycle status were detected by flow cytometry. And the expressions of cell cycle related proteins cyclin D1, cyclin B1 and CDK4 were detected by immunocytochemistry and Western blot. After TGM1 silencing, the HaCaT cells increased in G(0)/G(1) phase (78.27% ± 1.83% vs 56.84 % ± 2.72% and 57.19% ± 3.72%, both P < 0.05) while decreased in S phases (32.78% ± 5.48% vs 57.32% ± 2.91% and 55.71 % ± 2.84%, both P < 0.05) and G(2)/M phases (3.66% ± 0.30% vs 9.39% ± 0.68% and 9.77% ± 0.52%, both P < 0.05) . Moreover, arrest of cell cycle was induced in S phase. After TGM1 silencing, cell proliferation was significantly inhibited while wild-type TGM1 displayed an accelerated growth rate. Inmmnohistochemistry indicated that cyclin D1, cyclin B1 and CDK4 had a strongly negative expression after transfecting with TGM1 siRNA. Western blot indicated that a very low endogenous expression of cyclin D1, cyclin B1 and CDK4 proteins was observed after transfecting with TGM1 siRNA. TGM1 siRNA may block the cell cycle of HaCaT cells. It suggests that TGM1 expression in HaCaT cells are closely related with cell cycle.

Citation

Sanquan Zhang, Xinjing Gao, Wenyuan Qiu, Quan Chen, Xin Zhou, Xin Tian, Zhiping Tang, Tian Zhao, Fang Zhang, Wei Li, Xibao Zhang. Mechanism and influence of silencing TGM1 gene by siRNA on cell cycle distribution of keratinocytes]. Zhonghua yi xue za zhi. 2014 Dec 2;94(44):3478-82

Mesh Tags

Substances

PMID: 25622737

search → result