实用肝脏病杂志Issue(5)：523-526,4.DOI:10.3969/j.issn.1672-5069.2014.05.019
Rac1对 L02肝细胞株上皮细胞间质转化及增殖和凋亡的影响
Effects of Rac1 on epithelial-mesenchymal transition,and proliferation and apoptosis of L02 cells in vitro
摘要
Abstract
Objective To investigate the effects of Rac1 on epithelial mesenchymal transition(EMT) induced by transforming growth factor β1(TGF-β1),and on cell proliferation and apoptosis of L02 cells in vitro. Methods Rac1 plasmids with different activity including pExRed-NLS Flag(vector),pExRed-NLS Flag Rac1(wild type),pExRed-NLS Flag Rac1T17N (dominant negative mutant) and pExRed-NLS Flag Rac1G12V(constitutively active mutant)were transiently transfected into L02 cells,followed by stimulation of exogenous TGF-β1 at dose of 5ng/ml;Exogenous Flag -Rac1 fusion protein was determined by Western blot analysis;Immunofluorescence and Western blot were used to evaluate epithelial markers of Ck8 and mesenchymal markers of vimentin;Cell motility was assessed by transwell assay and wound healing assay;L02 cells were treated with different concentrations of Rac1 inhibitor(NSC23766),and the influence of Rac1 on cell proliferation and apoptosis were detected by CCK-8 assay or Annexin V -FITC/PI double staining,respectively. Result All kinds of plasmids were successfully transiently transfected into L02 cells;Compared with the vector group and the wild type group,constitutively active mutant pExRed-NLS Flag Rac1G12V significantly increased the expression of vimentin,decreased the expression of CK8 and enhanced cell motility,whereas the effects of dominant negative mutant pExRed-NLS Flag Rac1T17N were just the opposite of above results (P ﹤0.05);Disruption of Rac1 activity with NSC23766 inhibited cell proliferation (P﹤0.05) without increasing cell apoptosis. Conclusions Rac1 promotes the EMT process and cell proliferation induced by TGF-β1 in L02 cells without affecting cell apoptosis in vitro.关键词
L02细胞/Rac1/上皮间质转化/增殖/凋亡Key words
L02 cells/Rac1/Epithelial-mesenchymal transition/Proliferation/Apoptosis引用本文复制引用
王晖晖,张峰,沈珊珊,诸葛宇征..Rac1对 L02肝细胞株上皮细胞间质转化及增殖和凋亡的影响[J].实用肝脏病杂志,2014,(5):523-526,4.基金项目
江苏省自然科学基金资助项目 （）
