PLVAP促进肝癌发展的分子机制研究

Previous studies demonstrated that, 5-fluorouracil combined with pegylated interferon alpha-2b in treating unresectable hepatocellular carcinoma prolongs patient survival (published), and a kind of new proteins PLVAP might increase the effect of combined chemotherapy significantly. However the significance of PLVAP in HCC, its biological function and molecular mechanism remains unknown. In preliminary experiments, we found that PLVAP is highly expressed in HCC, and it is associated with prognosis. Through stablishing stable cell lines of overexpressing and silencing PLVAP in Huh-7 cells, we found that PLVAP promoted cell proliferation, angiogenesis, inhibiting apoptosis, and bioinformatic analysis results further showed that NF-κB signaling pathway was activated in HCC patients with high expression of PLVAP. On this basis, we will further prove that PLVAP could be used as a biomarker for diagnosis and prognosis, and that PLVAP promotes the progression of HCC in mutiply HCC cell lines and in nude mice in vivo. Furthermore, through immunoprecipitation and other methods, we will prove the hypothesis that PLVAP increases the expression of TNFR in cell membrane, and binds to TNFR to form membrane complex, recruiting TRAF、RIP and promoting their K63 ubiquitilation, resulting in the activated NF-κB signaling pathway to promote the progression of HCC, providing new clues for the diagnosis and treatment of HCC.

前期研究证明5-Fu与聚乙二醇干扰素α-2b的联合用药显著提高肝癌病人预后（已发表），而新蛋白PLVAP可明显提高肝癌联合用药治疗效果。然而PLVAP在肝癌中的作用及分子机制还有待进一步研究。在预实验中，我们发现PLVAP在肝癌中高表达，其表达与预后相关。通过建立稳定高表达及沉默PLVAP表达的Huh-7细胞系，我们发现PLVAP促进细胞增殖、血管新生，抑制细胞凋亡，进一步通过生物信息学分析发现PLVAP激活NF-κB信号通路。在此基础上，本项目将进一步证明PLVAP可作为肝癌诊断、预后标志物，并在多个细胞系及裸鼠体内验证PLVAP可促进肝癌发展，通过免疫蛋白共沉淀法等阐明PLVAP促进细胞膜上TNFR聚集，与TNFR结合形成膜上复合物，招募TRAF2、RIP等，促进其K63位泛素化，激活NF-κB信号通路从而促进肝癌发展的新假说，为肝癌的诊断及治疗提供新的线索。

前期研究证明5-Fu与聚乙二醇干扰素α-2b的联合用药显著提高肝癌病人预后，而新蛋白PLVAP可明显提高肝癌联合用药治疗效果。然而PLVAP在肝癌中的作用及分子机制还有待进一步研究。在预实验中，我们发现PLVAP在肝癌中高表达，其表达与预后相关。通过建立稳定高表达及沉默PLVAP表达的Huh-7细胞系，我们发现PLVAP促进细胞增殖、血管新生，抑制细胞凋亡，进一步通过生物信息学分析发现PLVAP激活NF-κB信号通路。在此基础上，本项目进一步证明了PLVAP可作为肝癌诊断、预后标志物，并在多个细胞系及裸鼠体内验证PLVAP可促进肝癌发展，通过免疫蛋白共沉淀法等阐明了PLVAP促进细胞膜上TNFR聚集，与TNFR结合形成膜上复合物，招募TRAF2、RIP等，促进其K63位泛素化，激活NF-κB信号通路从而促进肝癌的发展，为肝癌的诊断及治疗提供新的线索。