V-ATPase V1G1促进肝癌转移的分子机理及作为转移预测分子的临床应用价值

肝癌发生发展的高酸微环境伴随V-ATPase的表达增加及活性增强。前期研究发现：pH调节蛋白V-ATPase V1G1亚基在肝癌细胞及肝癌组织中高表达，其在肝癌细胞的表达水平与细胞转移潜能呈正相关；用特异性V-ATPase抑制剂处理肝癌细胞，其生存率呈剂量依赖性下降；且不同pH培养基中的肝癌细胞生存率随pH的上升而下降；但其临床应用价值及作用机制均未阐明。据此提出假说：V1G1亚基可能是通过癌细胞中V1G1过表达－细胞外基质酸性pH加剧－肿瘤微环境的相关因素改变－肿瘤转移的方式促进肝癌转移，可作为肝癌转移的预测分子。本项目拟通过不同转移潜能的肝癌细胞系、肝癌组织标本、裸鼠移植瘤模型为研究对象，采用慢病毒转染、CRISPT/Cas9基因敲除技术、信号通路芯片、流式细胞术及免疫组化等技术，从分子、细胞、组织及动物等多个层次明确V1G1在肝癌转移中的作用及作为肝癌转移预测分子的临床价值。

The hepatocarcinogenesis and development accompany with the overexpression and high activity of V-ATPase. Our previous data shown that V-ATPase V1G1 was significantly increased in both HCC cell lines and human HCC tissue samples, and even more in HCC cell lines with high metastasis potential; besides, when treated with V-ATPase inhibitor, the survival rate of HCC cell lines was significantly decreased in a does dependent way; This effect was similar with that when cultured HCC cell lines in high pH medium, their survival rate was notably decreased; but the clinical value of V-ATPase V1G1 in predicting metastasis of Hepatocellular Carcinoma and its mechanism remains unknown. We therefore proposed a hypothesis that V-ATPase V1G1 might promote the HCC metastasis by V1G1 overexpression in HCC cells-increased acidic pH in extracellular matrix- changes in tumor microenvironment-HCC metastasis, and can be a potential biomarker for HCC metastasis. By using HCC cell lines, HCC patient samples and HCC xenograft mice model, we will elucidate the effects and predictive role of V-ATPase V1G1 on the HCC metastasis, as well as its mechanism in regulating acidic microenvironment by lentiviral transfection，CRISPT/Cas9 gene knockout technology，cell signaling microarray techniques，Flow cytometry，immunohistochemistry and so on. Our research will provide a novel V-ATPase V1G1 targeted method in treatment and prevention of HCC.