miR-29b调控卵巢癌细胞Warburg效应的作用及分子机制研究

Abnormal energy metabolism, especially the Warburg effect which is characterized by the shift in glucose intake from oxidative phosphorylation to glycolysis, is an essential indicator of malignant transformation. As important regulatory factors, miRNAs have become current research focuses in the field of carcinoma energy metabolism regulation. miR-29b has been reported to be dysregulated in a series of human carcinoma and involved in multiple steps of malignant progression. However, its role in Warburg effect and underlying mechanism are still on the waiting list. In our previous study, we found that miR-29b had two potential targets, both of which are key genes in the well acknowledged metabolism-related PI3K/AKT/mTOR signaling pathway. Therefore, we speculate that miR-29b may have a significant effect on the regulation of abnormal energy metabolism in ovarian carcinoma, which has not yet been studied worldwide. Based on the above assumption, this project aimed to: ① demonstrate the impact of miR-29b expression level on energy metabolism in ovarian carcinoma by using a variety of ovarian carcinoma cell lines and their models in nude mice; ② detect the expressions of miR-29b, and the two potential target genes ( AKT2 and AKT3) in ovarian carcinoma tissues and analyze their correlation with biological characteristics of ovarian carcinoma progression; ③ identify the regulatory relationship between miR-29b and the two potential target genes ( AKT2 and AKT3) via technologies such as the dual luciferase reporter assay system, the regulation of the miRNA expression, and the RNA and protein detection techniques, etc; ④ explore the influence of miR-29b on the PI3K/AKT/mTOR signaling pathway in the field of carcinoma energy metabolism and key enzymes involved in Warburg effect of ovarian carcinoma cells by further regulating miR-29b expression level. This project is supposed to lay a theoretical foundation for understanding the mechanism by which Warburg effect in ovarian carcinoma is regulated, and to provide new directions in the targeted therapy for ovarian carcinoma.

以有氧糖酵解为特征的Warburg效应是细胞恶性转变的重要特征。miRNAs对肿瘤代谢的调控是当前研究热点。miR-29b在多种肿瘤中表达异常并参与了肿瘤发生发展，但其在Warburg效应的作用及机制尚无报道。前期研究发现miR-29b有靶向调节AKT2/3的潜力，由此推测,miR-29b可能通过靶定AKT/mTOR通路而影响卵巢癌细胞Warburg效应。本项目拟：①利用多种卵巢癌细胞株及裸鼠荷瘤模型明确miR-29b对卵巢癌细胞能量代谢的影响；②检测卵巢癌组织miR-29b和AKT2/3的表达并分析其与肿瘤生物学特征的相关性；③采用报告基因、miRNA表达调控、RNA及蛋白检测技术验证miR-29b与靶基因的调控关系；④通过调控miR-29b表达检测其对AKT/mTOR通路及Warburg效应关键酶的影响。本项目将为探索卵巢癌Warburg效应的调控机制和靶向治疗提供理论参考和新思路。

研究背景：以有氧糖酵解为特征的Warburg效应是细胞恶性转变的重要特征。miRNAs对肿瘤代谢的调控是当前研究热点。miR-29b在多种肿瘤中表达异常并参与了肿瘤发生发展，但其在Warburg效应的作用及机制尚无报道。前期研究发现miR-29b有靶向调节AKT2/3的潜力，由此推测,miR-29b可能通过靶定AKT/mTOR通路而影响卵巢癌细胞Warburg效应。.主要研究内容：本项目①利用多种卵巢癌细胞株及裸鼠荷瘤模型明确miR-29b对卵巢癌细胞能量代谢的影响；②检测卵巢癌组织miR-29b和AKT2/3的表达并分析其与肿瘤生物学特征的相关性；③采用报告基因、miRNA表达调控、RNA及蛋白检测技术验证miR-29b与靶基因的调控关系；④通过调控miR-29b表达检测其对AKT/mTOR通路及Warburg效应关键酶的影响。.重要结果：受国家自然科学基金青年基金项目资助，本项目通过体内和体外实验证实：①miR-29b在不同卵巢癌细胞系及正常/病变卵巢组织中均存在差异性表达，miR-29b参与调解卵巢癌细胞的能量代谢；②miR-29b直接靶定并负性调节AKT2/3的表达水平；③AKT2/3的表达水平与卵巢癌的进展及糖酵解代谢存在显著的相关性；④miR-29b沉默所引起的AKT表达上调将进一步激活卵巢癌细胞中Warburg效应通路中关键分子的表达。⑤体内实验证实miR-29b过表达将有效抑制异位移植瘤的生长。.科学意义：本项目通过细胞水平、组织水平及动物水平的多项实验，证实了miR-29b—AKT2/3通路在卵巢癌细胞Warburg效应中的重要作用，为探索卵巢癌Warburg效应的调控机制和靶向治疗提供理论参考和新思路。