Wnt通路调控mir-106b、mir-93/RhoC通路影响卵巢癌侵袭转移的分子机制解析

Ovarian cancer is one of the most common gynecologic malignant tumors in which invasion and metastasis lead to poor prognosis. We have found that RhoC promoted ovarian cancer invasion and metastasis through participating in the EMT process. Though Wnt-5a could regulate the expression of RhoC, RhoC were not the direct target gene in the Wnt pathway. We also found that miR-93 and miR-106b were down-regulated in ovarian cancer tissue in our preliminary work. And after transfected with miR-93 and miR-106b respectively, cell proliferation and invasion ability were inhibited, RhoC protein expression was also down-regulated. According to the analysis of bioinformatics, there were combinations of miR-106b and miR-93 in the RhoC 3＇UTR area, whereas there were combinations between the transcription-regulation area of the miR-93 and miR-106b and the transcription factor E2F1 in the Wnt pathway. Through the reporter gene analysis, Chromatin immunoprecipitation (ChIP), cell proliferation, invasion, metastasis functional test in vitro, and tumor formation in nude mice, metastatic tumor formation experiment, we are trying to confirm that, the transcription factor E2F1 of Wnt pathway may regulates RhoC expression through regulating miR-93 and miR-106b, and participate in the invasion and metastasis of ovarian cancer.

卵巢癌是女性器官常见的恶性肿瘤。侵袭转移后是预后不良的重要原因。我们前期工作发现RhoC参与卵巢癌细胞上皮-间质转化（EMT）促进卵巢癌侵袭转移，Wnt-5a可调节RhoC的表达。但是RhoC不是Wnt通路的直接靶基因。我们还发现miR-93和miR-106b在卵巢癌组织中表达降低，在卵巢癌细胞中分别转染miR-93和miR-106b，细胞增殖抑制，侵袭能力降低，根据生物信息学分析发现RhoC的3'UTR区具有miR-106b和miR-93的结合位点，而wnt通路上的转录因子E2F1与miR-106b和miR-93的转录调控区有结合位点，因此我们拟通过报告基因分析、染色体免疫沉淀、体外细胞增值侵袭转移等功能试验及裸鼠成瘤试验及转移瘤形成等试验，力图证明：Wnt通路的转录因子E2F1通过调控miR-106b和miR-93进而调节RhoC表达，参与卵巢癌侵袭转移。

卵巢癌是女性器官常见的恶性肿瘤。侵袭转移后是预后不良的重要原因。我们发现miR-93、miR-106b和miR519d在卵巢癌组织中表达降低，在卵巢癌细胞中分别转染miR-93、miR-106b和miR519d，细胞增殖抑制，侵袭能力降低。根据生物信息学分析发现RhoC的3'UTR区具有miR-93、miR-106b和miR519d的结合位点，而wnt通路上的转录因子E2F1与miR519d的转录调控区有结合位点，因此我们通过报告基因分析、体外细胞增殖侵袭转移等功能试验及裸鼠成瘤试验及转移瘤形成等试验，证明了Wnt通路的转录因子E2F1通过调控miR-519d进而调节RhoC表达，参与卵巢癌侵袭转移。除此之外，我们还发现了多条参与卵巢癌发生发展的非编码RNA,例如：MicroRNA-490-3P，MicroRNA-372，LncRNA ABHD11-AS1， LncRNA TDRG1等。本研究的顺利实施，将丰富卵巢癌发生发展的理论研究，并可能为其阻断治疗提供新的靶点。