miR-224调控ASS1介导的能量代谢重编程在肝癌进展中的作用及机制研究

Warburg effect, the main way of energy metabolic reprogramming in tumor cells, plays an important role in tumor progression. Our previous study found that the expression of ASS1 was lower in tumor tissue than that in tissue adjacent, and negatively correlated with prognosis of HCC patients. We also found that the inhibition of ASS1 promoted the proliferation of HCC cells in vivo and in vitro experiments. Bioinformatics analysis indicated that ASS1 might participate in the regulation of energy metabolic reprogramming in HCC cells, and ASS1 expression might be regulated by miR-224. However, the function of ASS1 in HCC and its mechanism have not been reported. Thus, we speculated that the expression of ASS1 might be regulated by miR-224 and play a crucial role in HCC development by modulating the energy metabolic reprogramming. In our study, energy metabolism detection system and metabonomics technology were utilized to verify the role of ASS1 in HCC progression, and investigate the regulation and its mechanism of ASS1 in energy metabolic reprogramming of HCC via clinical tissue specimens, in vivo and in vitro experiments. We also explored the mechanism of low ASS1 expression regulated by miR-224 in HCC. In a word, this study will illustrate the mechanism of ASS1 involved in HCC progression and regulating mechanism of its low expression in HCC, which will provide novel ideas and strategies for the target treatment of HCC.

以“Warburg效应”为主要方式的能量代谢重编程在肿瘤进展中起到重要作用。我们前期研究发现：ASS1在肝癌中低表达并且和临床预后密切相关；ASS1能够调控肝癌细胞的增殖；生物信息分析提示：ASS1可能参与肝癌细胞能量代谢重编程的调控，其低表达可能受到miR-224靶向调节。但目前关于ASS1在肝癌进展中的作用及机制尚无相关报道。据此我们推测：miR-224调控ASS1的低表达，进而影响能量代谢重编程，促进肝癌进展。本课题拟从组织标本、细胞实验和小鼠荷瘤模型三个水平，利用能量代谢检测系统和代谢组学技术，系统地阐明miR-224/ASS1在肝癌进展中的作用；证实ASS1调控肝癌增殖的作用；确定ASS1对肝癌能量代谢重编程的调控及机制；探索ASS1低表达的上游调控机制；揭示miR-224调控ASS1介导的能量代谢重编程在肝癌进展中的作用及机制，为肝癌的靶向治疗提供新思路和新策略。

以“Warburg效应”为主要方式的能量代谢重编程在肿瘤进展中起到重要作用。我们研究发现：在肝癌组织中ASS1表达明显低于癌旁组织，但ASS1参与肝癌进展的机制及其异常表达的调控机制有待进一步阐明。本课题利用TCGA和GEO公共数据库分析ASS1的表达在肝癌中的预后判断价值，解析糖酵解、脂肪酸代谢通路在ASS1不同表达水平的富集情况，以及ASS1与糖酵解关键酶PKM2在肝癌组织中表达的相关性；利用生物信息学工具筛选并鉴定出能够在肝癌中靶向调节ASS1表达的miR-224，并分析其对于肝癌的预后判断价值，通过体内外实验进一步验证 miR-224对ASS1的表达调控。阐明ASS1通过调控PKM2的表达影响肝癌细胞的代谢重编程进而影响肝癌进展的作用机制及肝癌中ASS1低表达的调控机制，并希望以此找到新的靶点，为开发临床治疗药物奠定相关基础。