许多研究发现肝癌干细胞是影响肝癌复发转移及预后的重要因素，而肿瘤微环境对肝癌细胞的干性转化及维持起重要促进作用。申请人近期研究发现与活化的肝星状细胞共培养后肝癌细胞可获得干性表型，并筛选出了原代肿瘤相关成纤维细胞（CAFs）促肝癌细胞干性转化的关键基因SCUBE1。在本项目中，我们拟进一步从分子、细胞及动物水平验证SCUBE1基因是否对肝癌细胞干性转化及侵袭转移有促进作用；探讨SCUBE1/Shh/Gril1信号通路调控肝癌细胞干性转化的具体分子机制；并通过前瞻性队列利用参数及非参数方法构建联合SCUBE1水平与临床数据的肝癌术后转移复发预测模型。本研究的结果，将有助于阐明肝癌干细胞产生的可能机制及SCUBE1基因在其中发挥的关键作用，并且以SCUBE1表达水平联合临床数据构建的预测模型可应用于肝癌转移复发高危人群的早期筛检与预后评估，从而达到早发现、早诊断、早治疗的预防目标。

Many studies have found that liver cancer stem cells(LCSCs) is an important factor affecting the recurrence and metastasis of liver cancer, and the tumor microenvironment plays an important role in the process of liver cancer cells get stem cell phenotype. In our previous study, we found the liver cancer cells would get stem cell phenotype when they were co-cultured with activated hepatic stellate cell(LX2). Moreover, using gene chip, cancer associated fibroblasts(CAFs) primary culture and qPCR method, we screened a key gene, SCUBE1, which was produced by CAFs and could promote liver cancer cells transform into LCSCs. In this project, by constructing Lentivirus loaded SCUBE1 shRNA plasmid, co-culture experiment, orthotopic co-transplantation model of HCC in nude mice, we would verify the effect of SCUBE1 on the process of the liver cancer cells get stem cell phenotype in vivo and vitro. By using luciferase reporter system and co-immunoprecipitation technique, we will investigate the mechanism of how SCUBE1 control the process of liver cancer cells get stem cell phenotype. Through constructing prospective cohorts, we will establish the liver cancer recurrence and metastasis prediction model which combined the SCUBE1 with the clinical information by using parametric and nonparametric methods. The findings of this study will help to clarify the possible mechanism involved in the generation of LCSCs and the mechanism of SCUBE1 play a key role in the process of generation of LCSCs. Furthermore, the prediction model can be used to screen high risk patients who would have worse prognosis and then to achieve the early prevention final goal.