β1-integrin-mediated Protein Kinase C-α Activation Induces Nuclear Factor Kappa B Expression And Invasion, Metastasis Of Hepatoma Cells

ObjectiveHepatocellular carcinoma(HCC)is a threat to human life and health of one of the most important cancer.Outbreak mechanism and molecular biology mechanism of thorough study HCC will become decide its treatment success or not of important breakthrough.As for the survival of the local environment,extracellular matrix(ECM) and the transmission signal plays an important role to the maintenance of normal cell morphology and function and inducing the occurrence and development of tumor cells.β1-integrin family is considered mediated ECM signals in the main cell surface receptors,System researchβ1-integrin signal's turning to lead thoroughfare is doubtless the key which understands ECM to the HCC influence,is also the problem that researcher's many in the last years hopes resolve.Protein kinase C(PKC)and the regulation of transcription factor can be related to the promotion of tumor cell proliferation and inhibit apoptosis,and PKC-related transcription factor and the effectiveness of this regulation is important to a certain extent byβ1-integrin the impact, the project's goal is toβ1-integrin in the signal transduction pathway related to the activation of PKC isozymes on and positioning,from the cellular signal transduction perspective,further clearβ1-integrin signaling pathway in the cascade effect factor,and improve the response of the chain structure,to clarify the ECM to promote the development of HCC in the molecular level mechanism to provide the theoretical foundation. MethodspEGFP-N1 transient transfection plasmid and pEGFP-N1/GFβ1A transient transfection plasmid were successfully transfected into HepG2 cells by the cell transfection experiments.To observe the effect ofβ1-integrin on PKCαactivity, NF-κB expression and cell invasion of HepG2 cells and byβ1-integrin induced PKCαthe activation,NF-κB expression and Liver cancer cell invasiveness change situation under the Go6976 intervention in vitro invasion of Transwell Experimental,Western imprinting experiment,immunohistochemical staining experiments,electrophoretic mobility shift assay(EMSA)experiments and specific inhibitor of PKC Go6976 inhibition experiments.ResultspEGFP-N1 plasmid and pEGFP-N1/GFβ1A plasmid transfected HepG2 cells after 48h,The cells' growth were exuberant and assumed the obvious green fluorescence under the fluorescence microscope.Reverse transcriptase-polymerase chain reaction (RT-PCT)experiment certificates transfection' successfully.β1-integrin expression vector transfected HepG2 cells' invasion increased significantly,compared with the empty vector transfected HepG2 cells,the difference was significant(P＜0.05).Western blot test and immunohistochemical staining experiments showed that the promotion ofβ1-integrin activation of PKCαHepG2 cells,and to promote the PKCαmembrane to plasma translocation.EMSA results showed thatβ1-integrin promotes NF-κB activation of HepG2 cells.Go6976 significantly inhibitedβ1-integrin induced activation of PKCα,Simultaneously also inhibitβ1-integrin-mediated NF-κB activity and cancer cell invasion.Conclusionβ1-integrin-mediated activation of PKCαactivity,thereby induced NF-κB expression,leading to the invasiveness of hepatocellular carcinoma cells increase in the promotion of tumor cell invasion and metastasis.The discovery of this pathway,the significance is that it is conducive to deepen our understanding of HCC cells in theβ1-integrin signaling pathway cognitive structure,strengthening and development of the HCCβ1-integrin function of understanding,more importantly,ECM will help to completely clear signal in the development of HCC in the role of mechanisms to clarify the eventual development of HCC induced by the drivers and hub link has a profound significance and will also develop a treatment strategy and significantly improve HCC the therapeutic effect of opening up a new channel.