The Mechansm Of Mr-765 Promoting Proliferation,Invasion And Inhbiting Apoptosis Of Hepg2 Cells Through LHPP Was Studied

Objective Toinvestigatethe mecharism of miR-765promoting proliferation invasion and inhibiting apoptosis of HepG2 cells through LHPPMethod:1.The upstream target genes of LHPP were predicted by bioinformatics analysis and verified by luciferase reporter gene assay.2 Immunchist ochemistry was used to detect the expression of miR-765 and LHPP in the tissues of hepatic cell carcinama HCC and adjaoent normal tissues The expression levels of miR-765 and LHPP in tumor tissues,paracancerous tissues and serum of HCC hepat coellular patients were detected by qRT-PCR.The expresion levels of miR-765 and LI-PP in human hepat coellular carcinoma Hub-7,HepG2,Hep3B and human normal hepat ocytes L-02 were detected by qRT-PCR,and suitable cell lines were selected3.NC,miR-765mimic,miR-765inhibitor,pcDNA-LHPP,si-LHPP and miR-765mimic+ pcDNA-LHPP plasmid were constructed to up-regulate or down-regulate the expressicn of miR-765 and LI-PP in HepG2 oells,respectively.The effects of miR-765 and LI-PP expression on colony formation,cell prdiferation,invasicn and apoptcsis of HepG2 cells after treatment were aranlyzed by clonal colony formation compartment invasicn and flow cytometry.4.The effect of miR-765 expression on N-cacherin E-cacherinβ-caterin related to Epithelial mesenchymal transition(EMT)in tumor cells was detected by Western blot.Protein levls of Vimentin Snail.5.The relationship between serum miR-765 expression and dinical phenotypes in patients with HCC was further explored.Results 1.Biainformatics prediction showed that LHPP was the upstream target gene of miR-765,and luciferase reporter assay showed that miR-765 targeted to inhibit the transcriptional activity of LHPP.2 Immunchistochemistry showed that compared with adjacent normal tissues,LHPP protein expression was decreased in HCC tissues,while miR-765 was overexpressed in HCC tissues(P<0.05).Human hepat coellular carcinma HepG2 was confirmed as the study object by qRT-PCR.3.After transfection with miR-765 mimic and upregulation of miR-765,the donal formation prdiferation and invasion ability of HepG2 cells were sigrificantly enhanoed,while the apoptosis rate of HepG2 cells was decreased(P<0.05).After transfection with miR-765 inhibitor,the doring-formation ability,prodiferation and invasion ability of HepG2 cells were inhibited and the apoptosis rate was increased(P<0.05).Overexpression of LHPP could inhibit the formation of done,proliferation and invasion of HepG2 cells,and increase cell apoptosis.The ability of done formation proliferation and invasion of knockdown LHPP cells was sgrificantly enhanced,while the rate of apcptosis was decreased(P<0.05).When co-transfected with miR-765mimic+pcDNA-LHPP,the effects of miR-765 on done formation cell proliferation invasion ability and anti-apoptosis of HepG2 cells were weakened(P<0.05).4.Western blot showed that miR-765mimic could up-regulate the expression of N-cacherin,β-caterin,Vimentin,Snails positively ocrrelated with EMT,and down-regulate the expression of N-cacherin negatively ocrrelated with EMT(P<0.05).MR-765inhibitor was able to down-regulate the expression of N-cacherin,β-caterin,Vimentin and Snail proteins positively associated with EMT,and up-regulate the expression of E-cacherin protein negatively associated with EMT(P<0.05).5.Analysis of dirical sigrificance showed that high expression of miR-765 was associated with lymph mode metastasis and histdogical grade(P<0.05).Condusion:1.As an ancogenic factor of HCC,miR-765 is highly expressed in HCC patients and positively ocrrelatad wth the dinical stage of HCC patients,so it may be a potential biomaker of HCC.2 High expression of miR-765 can inhibit the expression of tumor suppressor gene LHPP and enhance the dcrirg prdiferaticn and invasicn ability of hepatoma cells The mechanismis to affect the expression of EMT related proteins N-cacherin,β-Catenin,vimentin,snail and E-cacherin.