Rap1b By Up-regulating The Expression Of Twist 1 To Enhance The Invasion And Migration Of HCC

ObjectiveTo investigate the function and molecule mechanism of Rap1 b in HCC.Methods1.The expression of Rap1 b was determined by using IHC and qRT-PCR in tumor tissue and paracancerous tissue.QRT-PCR and Western blot were used for measuring expression of Rap1 b in HCC cells.2.We constructed pcDNA3.1-Rap1 b vector and synthesised the Rap1 b inhibitor.Then the effects of Rap1 b on HCC proliferation were analyzed by MTS and colony formation assay.The effects of Rap1 b on HCC invasion and migration were detected by wound healing assays and transwell assays.3.QRT-PCR,Western blot and Dual-Luciferase reporter Assay were applied to identify the target genes of Rap1 b.4.Tumor growth assay was used to analyze the tumor formational ability in vivo.Tumor migration assay was used to analyze the tumor migrational ability in vivo.Results1.Rap1 b up-regulated in HCC tissues and HCC cells.2.Rap1 b promoted proliferation,migration and invision of HCC cells in vivo.3.Mechanism study revealed Rap1 b up-regulated Twist1 expression by enhancing its promoter activity.4.Rap1 b promoted tumor formation and migration of HCC cells in vitro.ConclusionThese findings suggested Rap1 b may effect the progression of HCC by targeting Twist1.