Coronin-1C/PSGL-1调控肿瘤细胞与血管内皮细胞黏附进而促进肝癌肺转移瘤形成的机制研究

肺是肝癌远处转移最常见器官，而肿瘤细胞与肺脏血管内皮细胞黏附，是肝癌肺转移瘤形成的关键。前期我们建立人肝癌高肺转移潜能细胞HCCLM9，筛选并首次发现Coronin-1C与肝癌肺转移瘤形成密切相关，同时研究表明黏附分子PSGL-1与Coronin-1C协同高表达。据此我们假设Coronin-1C可能通过调控PSGL-1增强肿瘤细胞与血管内皮细胞的黏附，促进肝癌肺转移瘤形成。本课题拟构建Coronin-1C的高表达及ShRNA的慢病毒表达载体，分别转染不同肺转移潜能肝癌细胞，采用功能化流动腔及体内实验检测Coronin-1C/PSGL-1介导的肿瘤细胞与肺血管内皮细胞黏附行为的改变，并进一步研究异常高表达Coronin-1C对PSGL-1 O-GlcNAc糖基化及硫酸化等修饰水平的影响，明确Coronin-1C/PSGL-1通路在肝癌肺转移瘤形成中的作用，为后续临床转化奠定理论基础。

Metastasis is a major cause of high mortality in HCC patients after surgical resection. Lung is the most frequent targets of HCC metastasis in humans. The hematogenous metastasis of tumors mainly includes three steps: the tumor cells pass through the vascular endothelial cells of the tumor tissues and move out of their original position, the tumor cells move via blood, and the tumor cells become implanted at the metastatic position. Adhersion between tumor cells and endothelial cells facilitates formation of experimental metastasis. With low and high metastatic potentials, the stepwise metastatic human HCC cells MHCC97L and MHCC97H, were established via repeated in vivo selection and characterized by a similar genetic background but with significant differences in spontaneous metastasis behavior. For a better insight into the characteristic organ site-specific tropism of HCC metastasis, lung-specific mtastatic human HCC cells HCCLM9, were established via repeated in vivo selection, providing appropriate model systems for providing appropriate model systems for comparative study on the molecular events correlated lung metastasis with HCC. HCCLM9 and MHCC97L cell were used for comparative membrane proteome profiling using SDS-PAGE and ESI-MS technologies. Candidate protein makers were further validated by Western blot on cells and IHC on animal tumor tissues. Coronin-1C was overexpressed in HCCLM9, and validated by western blot and IHC from nude mice tumor tissues, and could predict HCC invasive behavior. P-selectin glycoprotein ligand 1 (PSGL-1) encodes a protein critical for cell migration and chemotaxis, which prepare for the stable adhersion between tumor cells and endothelial cells. PSGL-1 was overexpressed in HCCLM9 validated by western blot. Here, to address the role of coronin-1C/PSGL-1 in lung-specific metastasis of HCC, we up-regulated its expression in MHCC97L using lentivirus-mediated overexpression, and down-regulated its expression in HCCLM9 cells using lentivirus mediated shRNA. The kinetic studies of tumor cells rolling adhesion medated by the ineraction of coronin-1C with PSGL-1 were performed with flow chamber system.The expression of coronin-1C is critical for PSGL-1 modification. The current work demonstrates coronin-1C facilitates tomor colonization through PSGL-1 mediated adhesion of tumor cells with endothelial cells.