基于CAFs来源的外泌体miRNA-20a介导PTEN/PI3K/AKT通路探讨加味龟鹿二仙胶汤干预非小细胞肺癌顺铂耐药机制

加味龟鹿二仙胶汤能够逆转非小细胞肺癌(NSCLC)顺铂化疗耐药，肿瘤相关成纤维细胞（CAFs）与NSCLC顺铂化疗耐药关系密切。前期研究证实A549细胞活化CAFs并促进miRNA-20a表达上调，进而CAFs来源的外泌体传递miRNA-20a促进NSCLC化疗耐药。进一步研究发现，加味龟鹿二仙胶汤能够抑制CAFs释放外泌体，调控PTEN/PI3K/AKT通路。因此，我们推测该方的增敏及逆转耐药作用与调控CAFs有关，提出“加味龟鹿二仙胶汤通过抑制CAFs释放外泌体，调节miRNA-20a介导的PTEN/PI3K/AKT信号通路以增敏及逆转化疗耐药”假说。本课题将构建气阴两虚型肺癌小鼠模型，通过体内和体外实验检测CAFs来源外泌体miRNA-20a介导的PTEN/PI3K/AKT信号通路以论证上述假说，为找到针对NSCLC化疗起到增敏及逆转耐药的药物和治疗方法提供科学依据。

The modified Guilu Erxian Jiao Formula can reverse cisplatin chemoresistance in non-small cell lung cancer (NSCLC).Cancer associated fibroblasts (CAFs) have a close relationship with cisplatin chemoresistance in NSCLC. Previous studies have confirmed that A549 cells can activate CAFs and promote the up-regulation of miRNA-20a, and then CAFs-derived exosomes transmit miRNA-20a to promote chemoresistance in NSCLC. Further study found that the modified Guilu Erxian Jiao Formula can inhibit CAFs to release exosomes and regulate PTEN/ PI3K / AKT pathway. Therefore, we speculated that the Formula’s effects of increasing drug sensitivity and reversing drug resistance were related to the regulation of CAFs. The hypothesis is as follows: modified Guilu Erxian Jiao Formula can regulate miRNA-20a-mediated PTEN / PI3K / AKT signaling pathway to sensitize and reverse chemoresistance by inhibiting the release of CAFs derived exosomes. In order to confirm this hypothesis, this project will build a lung cancer mouse model of deficiency of both qi and yin, and demonstrate the above hypothesis by detecting the miRNA-20a-mediated PTEN / PI3K / AKT signaling pathway in CAFs derived exosomes in vivo and in vitro. This project will provide a scientific basis for finding a drug and method to increase NSCLC chemosensitivity and reverse of drug resistance.