METTL3介导的m6A修饰调控AKR1C1表达在肝外胆管癌中的作用机制及临床价值研究

肝外胆管癌（ECC）发病机制中一个关键问题是“为何会出现胆管上皮细胞增殖与死亡的异常失调？”，这其中的调控机制迄今未明。我们前期通过m6A及转录本测序发现：ECC肿瘤组织中m6A水平明显升高、m6A修饰的关键分子METTL3的表达也显著增加、并与ECC的病程密切相关；而且METTL3的下游靶分子之一是AKR1C1基因，后者我们前期已证实能明确调节EEC细胞增殖与死亡。因此，我们推测METTL3介导的m6A修饰能通过调控AKR1C1的表达来影响ECC细胞增殖与死亡，并拟通过荧光素酶报告基因、干扰/过表达、多核糖体分析、细胞死亡抑制剂、及体内、外功能获得/缺失实验来探讨METTL3/m6A/AKR1C1轴在ECC中调控细胞增殖、死亡及影响病程的作用机制，评估METTL3/AKR1C1对ECC病情监测及预后判断的临床检测价值；为回答上述ECC关键问题、寻找ECC的潜在诊疗靶点提供思路借鉴。

A key question in the pathogenesis of extrahepatic cholangiocarcinoma (ECC) is "Why is there a dysregulation of proliferation and death in bile duct epithelial cell?". The regulation of this abnormal in ECC is still unknown. Through m6A sequencing and transcript sequencing, we found that m6A levels in ECC tumor tissues were significantly increased. At the same time, the expression of the key molecule METTL3 modified by m6A was significantly increased, and it was closely related to the course of ECC. AKR1C1 gene is one of the downstream target molecules of METTL3. We have previously confirmed that AKR1C1 can regulate the proliferation and death of EEC cells. Accordingly, we proposed that METL3-mediated m6A modification can affect the proliferation and death of ECC cells by regulating the expression of AKR1C1. To confirm this hypothesis, we will explore molecular mechanisms of METTL3/m6A/AKR1C1 axis and its regulation of the proliferation and death of ECC cells in vivo and in vitro, by combination of various experiments, including luciferase report, inhibition and over-expression, polysome profiling, cell death inhibitors, and so on. To evaluate the clinical detection value of METTL3/AKR1C1 for ECC monitoring and prognosis judgment, which may provide new ideas for answering the above key questions of ECC and finding a potential diagnostic or therapeutic target of ECC.