Study On The Effect Of Hic-5 In The Hepatic Stellate Cells To Liver Cell Proliferation

Objective: Hic-5 in Hepatic stellate cells were discussed from cytology level about effects on liver cell proliferation.Methods : In vitro culturing liver cell lines HL-7702 and hepatic stellate cell lines LX-2;The experiment is divided into three groups:A.people liver cell lines separate training group;B.Human hepatic stellate cells and liver cell lines co-culture group;C.Hic-5si RNA human hepatic stellate cells and liver cells co-culture group.For experimental hepatic stellate cell line with Hic-5 gene silencing,respectively,to establish each hepatic stellate cells and liver cells co-culture system in vitro.Trial and error to select training time node 0 h,24 h,48 h,72 h.With immunofluorescence technique to detect Hic-5,?-SMA protein expression for identification of activated hepatic stellate cells and the expression,the distribution of of Hic-5.With western blot technique to detect Hic-5,Collagen I,cyclind1 protein expression situation in different groups of different time points,and to detect the change of Hic-5 gene as the change of culturing time affecting the Collagen I synthesis and liver cellproliferation.With immunohistochemical to detect the expression of ki67 change of each different time points between groups USing CCK-8 to test cell survival and drawing cell proliferation curve,in order to explore the effects on liver cell proliferation of Hic-5.Result:(1)cell immunofluorescence measured Hic-5's expression in cultured human HSCs,and with ?-SMA expression in HSCs,with theactivation of HSCs,Hic-5 expression gradually enhanced.(2)Westernn bio method measured,as the set point of time i increasing in the same group,Cyclin D1,CollagenI expression levels are both rising.But at different time points within the Hic-5SiRNA human hepatic stellate cells and liver cells co-culturing groups expression of Cyclin D1 level are more than human hepatic stellate cells and liver cell lines co-culturing group,Hic-5siRNA human hepatic stellate cells and liver cells co-culturing groups express CollagenI levels are less than human hepatic stellate cells co-culturing group.(3)the Cell Counting Kit(CCK 8)method measured that the human liver Cell proliferation is significantly increased in Hic-5SiRNA human hepatic stellate cells and liver cells co-culture groups than another.Immunohistochemical method measured the Hic-5SiRNA human hepatic stellate cells and liver cells co-culturing groups expression of Ki-67 positive cells was more than human hepatic stellate cells and liver cell lines co-culturing group,and with the increasing of incubation time,the number of Ki-67 positive cells was also increasing.Conclusion:(1)the Hic-5 expression on HSCs,with the activation of HSCs,Hic-5 expression are gradually enhanced;(2)The liver cells is alive,with the increasing of incubation time,cell proliferation activity increases,with increased Cyclin d1 protein synthesis.(3)Activated human hepatic stellate cell promote thesecretion of HGF,and probably regulated by Hic-5.(4)Hic-5siRNA could reduce the synthesis of Collagen I or promote their decomposition.Hic-5SiRNA could promote liver cell proliferation.