Effects And Mechanisms Of Downregulated Expression Of HDGF On The Proliferation,Migration And Invasion Of Prostate Cancer Cells

ObjectiveTo study the effect and mechanism of hepatoma-derived growth factor(HDGF)on cell proliferation,migration and invasion of prostate cancer.MethodsLentivirus vectors were constructed according to the target sequence of human HDGF gene,which were then packaged and purified into lentivirus particles to transfect prostate cancer cells DU 145,PC3 and LNCaP.The cells were divided into three groups separately:?PBS:control group which was treated only by PBS,?shRNA-CTR:control group which was treated by lentivirus without target sequence,?shRNA-HDGF:experimental group which was treated by lentivirus with the target sequence.Relative mRNA expression of HDGF was evaluated by quantitative real-time polymerase chain reaction(qRT-PCR).CCK-8 assay and colony formation assay were performed to measure cell proliferation ability respectively.Cell migration and invasion capabilities were determined by scratch assay,migration assay and Matrigel invasion assay.Relative expressions of HDGF,phosphatidylinositol 3-kinase(PI3K),phospho-serine/threonine protein kinase(P-AKT),phosphorylated mammalian target of rapamycin(P-mTOR),epithelial-mesenchymal transition(EMT)-related proteins including E-cadherin,N-cadherin,Vimentin,Snail and Slug,and matrix metalloproteinases(MMPs)including MMP2 and MMP9 were examined by western blotting.ResultsLentivirus vectors were successfully constructed,packaged and stably transfected into prostate cancer cells DU 145,PC3 and LNCaP.QRT-PCR and Western blotting revealed that the mRNA and protein expression of HDGF in experimental group of prostate cancer cells DU145,PC3 and LNCaP were all downregulated(P<0.05).CCK-8 assay of DU145 and LNCaP cells and colony formation assay of DU 145 and PC3 cells indicated that cell proliferation of experimental group was inhibited(P<0.05).DU 145 and PC3 cells scratch assay and migration assay of DU145,PC3 and LNCaP cells revealed that cell migration capability of prostate cancer was inhibited(P<0.05).Matrigel irnvasion assay of DU 145,PC3 and LNCaP cells suggested that cell invasion capability of prostate cancer was reduced(P<0.05).Further more,Western blotting revealed that relative protein expression of PI3K,P-AKT and P-mTOR were all downregulated(P<0.05),and the expression of EMT-related proteins N-cadherin,Vimentin,Snail and Slug were down-regulated(P<0.05),while E-cadherin was up-regulated(P<0.05).In addition,matrix metalloproteinases(MMPs)including MMP2 and MMP9 were decreased after HDGF knockdown(P<0.05).ConclusionCell proliferation,migration and invasion in vitro of proatate cancer were inhibited by the downregulation of HDGF expression,which may be related with the inhibition of PI3K-AKT-mTOR signaling way,EMT as well as MMP2 and MMP9.It suggested that HDGF was a relevant protein in the progression of prostate cancer and may probably serve as a potentially therapeutic target for prostate cancer as well as its downstream argets.