title: [Mechanism and influence of silencing TGM1 gene by siRNA on cell cycle distribution of keratinocytes].
abstract: OBJECTIVE
  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).
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  TGM1 siRNA may block the cell cycle of HaCaT cells. It suggests that TGM1 expression in HaCaT cells are closely related with cell cycle.
authors.affiliation: Guangzhou Institute of Dermatology, Guangzhou 510095, China.
authors.forename: Sanquan
authors.initials: S
authors.lastname: Zhang
authors.forename: Xinjing
authors.initials: X
authors.lastname: Gao
authors.forename: Wenyuan
authors.initials: W
authors.lastname: Qiu
authors.forename: Quan
authors.initials: Q
authors.lastname: Chen
authors.forename: Xin
authors.initials: X
authors.lastname: Zhou
authors.forename: Xin
authors.initials: X
authors.lastname: Tian
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Tang
authors.forename: Tian
authors.initials: T
authors.lastname: Zhao
authors.forename: Fang
authors.initials: F
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Email: zxibao@126.com.
authors.forename: Xibao
authors.initials: X
authors.lastname: Zhang
mesh_terms: D015153:Blotting, Western; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002455:Cell Division; D002460:Cell Line; D049109:Cell Proliferation; D051358:Cyclin-Dependent Kinase 4; D005434:Flow Cytometry; D020868:Gene Silencing; D006801:Humans; D015603:Keratinocytes; D034622:RNA Interference; D034741:RNA, Small Interfering; D014162:Transfection; D011503:Transglutaminases
chemical_list: D018797:Cell Cycle Proteins; D034741:RNA, Small Interfering; D011503:Transglutaminases; C105804:transglutaminase 1; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4