miR-200家族对胆管癌干细胞自噬的影响及其机制研究

As the malignant core components of cholangiocarcinoma,cholangiocarcinoma stem cells have special autophagy mechanism, which may result in tumor development. The research of autophagy of cholangiocarcinoma stem cells could demonstrate the mechansim of cholangiocarcinoma tumourigenesis and provide the theoretical foundation of molecular treatment of cholangiocarcinoma.Our previous research of isolation and evaluation of cholangiocarcinoma stem cells have demonstrated that miR-200 plays an important role in cholangiocarcinoma stem cells, and that miR-200 could regulate the autophagy of cholangiocarcinoma, which prompts the pivotal regulated role of miR-200 family on autophagy of cholangiocarcinoma stem cells.In order to clarify that hypothesis,the objective of our research is to investigate correlation between the autophagy features and CSCs properties of cholangiocarcinoma stem cells ,to clarify the influence of miR-200 on cholangiocarcinoma stem cells by construct miR-200 expression vehicle,to demonstrate the mechanisms via which miR-200 regulates the autophagy of cholangiocarcinoma stem cells by construct experimental models in vivo and vitro, to clarify the effect point of miR-200 on autophagy of cholangiocarcinoma stem cells by dual-luciferase report assay, and to demonstrate the downstream pathway and mechansim of target gene of miR-200 family by lentivirus-based transfection.The expectant achievement of our research may demonstrate the autophagy characteristics and molecular mechanism of cholangiocarcinoma stem cells and lay a foundation for the improvement on molecular treatment effect of cholangiocarcinoma.

胆管癌干细胞是胆管癌恶性核心细胞，其所具有的特殊自噬机制，成为导致肿瘤的发生和发展的重要原因之一。探讨胆管癌干细胞的自噬现象和机制，对胆管癌治疗具有重要意义。我们前期分选、鉴定到胆管癌干细胞,并发现miR-200家族在胆管癌干细胞中发挥了极为重要的作用，并初步证实miR-200家族能调控胆管癌的自噬。本项目拟探讨胆管癌干细胞特性与其自噬现在的相关性；通过miR-200表达载体，验证其对胆管癌干细胞自噬的影响；通过构建体内外实验模型，验证miR-200家族在调节胆管癌干细胞自噬的作用位点；此外，通过构建双荧光素报告基因系统及慢病毒载体，验证miR-200家族在调节胆管癌干细胞自噬过程中的具体机制及下游通路。本项目有望进一步阐明胆管癌干细胞的自噬特性和分子机制，为分子靶向治疗提供新思路

胆管癌干细胞是胆管癌恶性核心细胞，其所具有的特殊自噬机制，成为导致肿瘤的发生和发展的重要原因之一。探讨胆管癌干细胞的自噬现象和机制，对胆管癌治疗具有重要意义。通过本项目的研究，我们发现胆管癌干细胞具有不同于非干细胞的自噬特性，miR-200b可以通过增加胆管癌细胞凋亡，减少5-氟尿嘧啶诱发自噬产生的保护作用，从而起到化疗增敏作用。同时我们发现另一条胆管癌差异性microRNA——let-7c，其可以通过EZH2和DVL3/β-catenin轴在胆管癌发生和远处转移过程中起到重要的调节作用。本项目进一步阐明了胆管癌干细胞的自噬特性、特异性microRNA生物学效应及其分子机制，为胆管癌后续的综合治疗提供了新思路