NF-κB激活gp96转录从而促进肝细胞生长、细胞周期进展和细胞转化

NF-κB-induced gp96 up-regulation promotes hepatocyte growth，cell cycle progression and transition

摘要:

摘要:【目的】探讨乙肝感染中gp96上调的机制及其可能发挥的病理学机制。【方法】首先通过生物信息学、Real-time PCR、荧光报告基因和Western blot研究NF-κB激活gp96表达的机制。进一步通过在肝细胞中过表达或敲低gp96的水平，运用CCK-8法和流式检测分析gp96对肝细胞增殖、凋亡，和细胞周期的影响，通过检测肝细胞EMT发生和细胞集落形成实验，分析gp96对于HCC发生的作用。【结果】NF-κB与gp96启动子上NF-κB结合位点结合，激活gp96的表达。实验结果显示，gp96能够促进肝细胞增殖、抑制凋亡，促进细胞周期从静息期向分裂期的转化，同时促进肝细胞EMT发生和细胞集落的形成。【结论】NF-κB通过活化gp96启动子上调其表达，为HBV慢性感染上调gp96的机制提供了线索，同时提示gp96在慢性炎症引发HCC过程中发挥重要作用。

Abstract:

Abstract:［Objective］To investigate the mechanism of gp96 raised during hepatitis B virus (HBV) infection and the pathological mechanism． ［Methods］ The mechanism of NF-κB activating gp96 expression was determined by bioinformatics analysis，luciferase reporter assay，real-time PCＲ and Western blot．The effect of over-expression and knockdown gp96 expression by transfection or ＲNA interference on hepatocyte proliferation，apoptosis and cell cycle was examined by CCK-8 and flow cytometry．The role of gp96 for HCC development was determined by epithelial-mesenchymal transition (EMT) and colony formation assay．［Results］NF-kB significantly increased the gp96 expression by binding to the NF-kB binding site．Over-expression and knockdown studies both show that gp96 promoted hepatocyte proliferation，inhibited apoptosis，and induced G0/G1 to S phase cell cycle progression．Moreover，gp96 induced epithelialmesenchymal transition and increased colony formation ability of hepatocytes．［Conclusion］Our results therefore provide insights in chronic HBV infection-induced gp96 expression，and indicate that elevated gp96 may contribute to HCC development during chronic inflammation．