研究显示肝癌CD133阳性细胞具备肿瘤干细胞特征，是临床治疗的靶标。我们前期发现高尔基体蛋白SCYL1BP1 在肝癌中低表达且与肿瘤分化程度正相关；该基因在肝癌CD133阳性细胞中表达明显低于CD133阴性细胞；敲减CD133阴性细胞SCYL1BP1表达可致OCT4表达上调、细胞增殖能力增强。提示该基因可能负调控肝癌CD133阳性细胞的生物学特性，但其机制不明。本项目拟首先应用基因导入或敲减技术，分别观察SCYL1BP1导入或沉默后对肝癌CD133阳性或阴性细胞自我更新、分化、增殖和体内成瘤等肿瘤干细胞特征的影响；进一步通过RNA-seq和免疫共沉淀等蛋白质组学技术，探讨筛选该基因导入或沉默后涉及的特异性信号分子及其通路，并在大样本肝癌组织芯片中验证；最后在肝脏条件性SCYL1BP1基因敲除小鼠模型中，确证该基因在肝癌发生发展中的重要作用。项目的实施有望为肝癌的肿瘤干细胞治疗提供新思路。

It was reported that CD133 positive hepatocellular carcinoma (HCC) cells had the characteristics of cancer stem cells (CSCs) and may be the target of future tumor therapy. Our previous study showed that SCYL1BP1, a golgin, was down-regulated in HCC samples. Expression of SCYL1BP1 was positively associated with differentiation grade of HCC. Futhermore, the expression level of SCYL1BP1 was much lower in CD133+ HCC cells than that in CD133- HCC cells. Knocking down SCYL1BP1 in CD133- HCC cells upregulated OCT4, a critical gene in stem cell self-renewal; and promoted tumor cell proliferation. These data suggested that SCYL1BP1 negatively regulated biological features of CD133+ cells, while the detailed molecular mechanism remains unclear. In this study, we will try to conditionally overexpress/knockdown SCYL1BP1 in CD133+/- HCC cells, and investigate the effect on characteristics of CSCs both in vitro and in vivo, such as self-renewal, differentiation, and proliferation. Furthermore, by using RNA-seq and co-immunoprecipitation based-proteomics, we will screen the potential down-stream signaling pathways which are altered both in overexpressing and in knocking-down experiments and verified by immunohistochemistry in HCC tissue microarray. Finally, we will investigate the role of SCYL1BP1 in HCC carcinogenesis by establishing and using liver conditional SCYL1BP1 knockout mice model. Hopefully our research will offer a new way for future HCC CSC therapy.