Preliminary Studies On Heparanase Mediated Invasion And Migration Of Hepatocellular Carcinoma Via The ?-catenin/MMP7 Pathway

Objective:To explore the biological function of heparanase(HPSE)in the invasion and migration of hepatocellular carcinoma,and to reveal the mechanism of potential function about relation between HPSE and ?-catenin/MMP7 pathway.Methods: In this experiment,HPSE RNAi cells were used to transfection(Lv)Hep G2 cell lines as experimental cell lines.The q RT-PCR and Western blot technologies were used to detect HPSE transfection cells gene transcription and protein expression level.Using the tablet cloning experiments to determine HPSE effects on Hep G2 cells proliferation.The scratches and transwell experiments were used to detect cell migration and invasion ability.The q RT-PCR and Western-blot technologies were used to detect the gene transcription and protein expression of pathyway and epithelial mesenchymal transition(EMT)related factors,that is,?-catenin,MMP7,E-cadherin and Vimentin.HPSE is validated by experiment within a nude mice into tumors had effects on hepatocellular carcinoma cell.Results: 1.Of three groups HPSE RNAi,Lv HPSE transfected Hep G2 cells showed that the m RNA transcription of HPSE in interference groups were significantly lower than that in mock group(p <0.05);and Western-blot showed that the protein expression of HPSE in interference groups were significantly lower than that in mock group.2.The results of clone formation assay showed that the proliferation rate in HPSE interference group was significantly lower than that in mock group(p <0.05).3.Scratch test results showed that HPSE interference group wound healing distance shortened,but no significant difference compared with the mock group.The results of transwell assay showed that the invasion and migration ability of HPSE in interference group was significantly lower than that in mock group(p <0.05).5.q RT-PCR results showed that: HPSE down-regulation can significantly change the m RNA levels of ?-catenin and MMP7,in addition,regulation of EMT related gene transcription,that is,up-regulation of E-cadherin,down-regulation of Vimentin.6.Western-blot results showed that ?-catenin and MMP7 proteins levels were impacted by down-regulation of HPSE,the experiment also found that down-regulation of HPSE also lead to up-regulation of E-cadherin and down-regulation Vimentin,which were the EMT-related proteins.7.The inhibitory effect of HPSE on HCC cells was confirmed by in vitro tumorigenicity experiment in nude mice,and the results were statistically significance(p <0.05).Conclusion: Down-regulation of HPSE can inhibit the proliferation,migration and invasion of Hep G2.Down-regulation of HPSE can inhibit the proliferation,migration and invasion of Hep G2 via may regulate ?-catenin/MMP7 signaling pathway.At the same time,down-regulation of HPSE can inhibit the invasion of Hep G2 by suppressing the EMT.