SREBP-2调控RAMP3基因表达在肝癌中的作用和机制研究

Hepatocellular carcinoma (HCC) is largely the result of genetic disorder induced by chronic liver disease. Its pathogenesis is complicating and unclear. Studies on new abnormal genes emerged in HCC closely related to conquest of this cancer. RAMP3 induces tumor cell glycolysis inhibition and promotes tumor cell apoptosis, yet its role in HCC remains unclear. Transcription factor SREBP-2 promotes occurrence and development of metabolic chronic liver disease is closely related to HCC pathogenesis. Based on previous analyses and large clinical sample sequencing data, we found that RAMP3 was down regulated significantly in HCC and negatively correlated with the prognosis of patients, which is a potential tumor suppressor gene. However, its regulatory mechanism and function need further research. According to current study, SREBP-2 is high expressed in HCC, and silence SREBP-2 can up regulate the expression of RAMP3, so RAMP3 is a potential transcription factor. The mechanism needs further study. This study based on the previous results, proposed to establish SREBF-2 as well as RAMP3 knockout and overexpression cell and animal models. And then using ChIP gene interference and editing technology in vitroivo, Luciferase reporter gene system, proteomics technology, bioinformatics, molecular biology and animal experiments to intensely research molecular signal network between SREBF-2 and RAMP3.

肝细胞癌（HCC）大多是肝细胞在慢性肝病作用下多基因失调积累并驱动的结果，发病机制复杂且不明确；基于机制研究，阐明在HCC发病多个环节起关键作用的异常基因是攻克HCC的关键。RAMP3抑制肿瘤细胞糖酵解，促进肿瘤细胞凋亡；SREBP-2转录因子促进代谢类慢性肝病的发生和进展，与HCC发病密切相关。前期基于临床大样本测序数据的分析和实验证实：RAMP3在HCC中被显著下调，与病人预后负相关，是潜在的抑癌基因；SREBP-2在HCC中高表达，在细胞中沉默SREBP-2能上调RAMP3的表达，是RAMP3潜在的调控因子，但具体调控机制与功能需进一步深入研究。本课题基于以上结果，拟建立SREBF-2和RAMP3基因沉默和过表达细胞和动物模型，在体内外综合运用ChIP，基因编辑，Luciferase报告基因系统，组学，生物信息学和动物实验深入研究SREBF-2调控RAMP3基因在HCC中的作用机制。

肝细胞癌（HCC）大多是肝细胞在慢性肝病作用下多基因失调积累并驱动的结果，发病机制复杂且不明确；基于机制研究，阐明在HCC发病多个环节起关键作用的异常基因是攻克HCC的关键。本课题实施建立了SREBF-2和RAMP3基因沉默和过表达模型，综合运用基因编辑，分子生物学和动物实验阐述了SREBF-2能够负调控RAMP3基因的表达；在体外过表达RAMP3基因能抑制肝癌细胞的增殖与侵袭等重要HCC恶性化特征，敲低SREBF-2基因能抑制肝癌细胞增殖与侵袭；在体内，过表达RAMP3的细胞成瘤性比敲除RAMP3基因的肝癌细胞差，以上结果证实了SREBF-2/RAMP3基因在肝癌中的功能，为下一步靶向治疗研究提供了一定的参考。