The Effects Of Fn14 On Metastasis In Human Epithelial Ovarian Cancer Cells

Objective: To compare the difference in Fn14 expression between HO8910 and HO8910-PM cells and to investigate the role of Fn14 in metastasis of human epithelial ovarian cancer cells and its possible mechanism Methods:1.The ability of migration and invasion were accessed using transwell assay 2.The protein expression of Fn14 in ovarian cancer cell lines HO8910 and HO8910-PM was detected by western blot 3.The Fn14 protein expression in HO8910 and HO8910-PM was detected by cell immunofluorescence.4.Construct plasmid which carries Fn14 gene,and transiently transfer it into HO8910-PM cells with lipofectmin 2000 in vitro and to detect the transfection efficiency 5.The ability of migration and invasion were accessed using transwell assay 3.The ability of proliferation was assayed by CCK-8 kit 6.The changes of the expression of Slug,E-cadherin,N-cadherin,vimentin,MMP2 and MMP9 were detected by Western blot.7.Construct HO8910-PM cells stably expressed of Fn14 gene 8.Transfer pLVX-Slug plasmid into HO8910-PM-Fn14 cells with lipofectmin 2000 in vitro and to detect the transfection efficiency 9.The ability of migration and invasion were accessed using transwell assay 10.The changes of the expression of Slug,E-cadherin,N-cadherin,vimentin,MMP2 and MMP9 were detected by Western blot.Results: Both migration and invasion number of HO8910-PM cells were higher than HO8910 cells.Compared with HO8910 cells,the expression of Fn14 in HO8910-PM was decreased obviously.Green fluorescent protein was seen in HO8910-PM cells,and expression level of Fn14 was significantly increased in pcDNA3.1-Fn14 transfected HO8910-PM cells.Overpression of Fn14 could decrease the ability of migration and invasion in HO8910-PM cells.When Fn14 gene was transferred,the protein expression of Slug,N-cadherin,vimentin,MMP2 and MMP9 were down-regulared,in contrast,the expression of E-cadherin was up-regulared.Green fluorescent protein was seen in HO8910-PM-Fn14 cells,and expression level of Fn14 was significantly increased compared with HO8910-PM cells.Red fluorescent protein was seen in HO8910-PM-Fn-14 cells,and expression level of Slug was significantly increased in pLVX-Slug transfected HO8910-PM-Fn14 cells.Overexpression of Slug could increase the ability of migration and invasion in HO8910-PM-Fn14 cells.When Slug gene was transferred,protein expressions of N-cadherin,vimentin,MMP2 and MMP9 were up-regulared,in contrast,protein expressions of E-cadherin was down-regulared.Conclusions: A higher ability of metastasis belonged to HO8910-PM cells may be correlated with decreased Fn14 expression.Fn14 could inhibit epithelial ovarian cancer cells metastasis and reverse EMT via down-regulating slug...