目的 探讨消岩汤逆转肺癌顺铂耐药的具体分子机制。方法 高表达Beclin1肺腺癌耐药（lung adenocarcinoma drug-resistant，A549/DDP）、低表达Beclin1 A549/DDP、高表达Yes关联蛋白1（Yes-associated protein 1，YAP1）A549/DDP和低表达YAP1 A549/DDP细胞分别给予消岩汤和顺铂干预，采用Transwell和克隆形成实验检测细胞侵袭和增殖；采用Western blotting检测Beclin1、YAP1、P糖蛋白（P-glycoprotein，P-gp）、肺耐药相关蛋白（lung resistance-related protein，LRP）、转录调节因子（transcriptional coactivator with PDZ-binding motif，TAZ）、磷酸化信号转导与转录激活因子3（phosphorylated signal transducers and activators of transcription 3，p-STAT3）、哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）表达。BALB/c小鼠sc高表达Beclin1 A549/DDP、低表达Beclin1 A549/DDP、高表达YAP1 A549/DDP和低表达YAP1 A549/DDP细胞建立移植瘤模型，ig消岩汤和瘤内注射顺铂治疗14 d，采用免疫组化检测肿瘤Beclin1、YAP1、P-gp和LRP蛋白表达。结果 消岩汤联合顺铂作用于高表达Beclin1和高表达YAP1肺癌细胞株，细胞侵袭和增殖能力增强（P＜0.05），而低表达Beclin1和低表达YAP1肺癌细胞株，肺癌耐药细胞株侵袭和增殖能力减弱（P＜0.05）；低表达Beclin1肺癌细胞株Beclin1、YAP1、P-gp和LRP蛋白表达水平均明显降低（P＜0.05）；高表达Beclin1肺癌细胞株Beclin1、YAP1、P-gp和LRP蛋白表达水平均明显升高（P＜0.05）；低表达YAP1肺癌细胞株p-STAT3、TAZ、Beclin1、YAP1、P-gp和LRP蛋白表达水平均明显降低（P＜0.05）；高表达YAP1肺癌细胞株TAZ、Beclin1、YAP1和P-gp蛋白表达水平均明显升高（P＜0.05）。小鼠移植瘤模型中，高表达Beclin1组和高表达YAP1组瘤体大于对照组，而低表达Beclin1组和低表达YAP1组瘤体小于对照组。结论 消岩汤通过Beclin1直接影响YAP1表达，消岩汤又可通过YAP1-TAZ通路影响Beclin1表达，消岩汤可能通过干预Beclin1-YAP1环路逆转肺癌顺铂耐药。

Objective To explore the mechanism of Xiaoyan Decoction (消岩汤) on reversing cisplatin resistance in lung cancer. Methods Lung adenocarcinoma cells with high expression of Beclin1 drug-resistance (A549/DDP), low expression of Beclin1 A549/DDP, high expression of Yes-associated protein 1 (YAP1) A549/DDP and low expression of YAP1 A549/DDP were intervened by Xiaoyan Decoction and cisplatin respectively, Transwell and clone formation experiments were adopted; Western blotting was used to detect Beclin1, YAP1, P-glycoprotein (P-gp), lung resistance-related protein (LRP), transcriptional co activator with PDZ-binding motif (TAZ), phosphorylated signal transducers and activators of transcription 3 (p-STAT3) and mammalian target of rapamycin (mTOR) expressions. BALB/c mice were sc cells with high expression of Beclin1 A549/DDP, low expression of Beclin1 A549/DDP, high expression of YAP1 A549/DDP and low expression of YAP1 A549/DDP to establish transplanted tumor models. Mice were ig Xiaoyan Decoction and intratumoral injection of cisplatin for 14 d, Beclin1, YAP1, P-gp and LRP proteins expressions in tumor were detected by immunohistochemistry. Results Xiaoyan Decoction combined with cisplatin enhanced the invasion and proliferation of lung cancer cell lines with high expression of Beclin1 and high expression of YAP1 (P < 0.05), while the invasion and proliferation of lung cancer cell lines with low expression of Beclin1 and low expression of YAP1 were weakened (P < 0.05). The protein expression levels of Beclin1, YAP1, P-gp and LRP in lung cancer cell lines with low expression of Beclin1 were significantly decreased (P < 0.05). The protein expression levels of Beclin1, YAP1, P-gp and LRP in lung cancer cell lines with high expression of Beclin1 were significantly increased (P < 0.05). The expression levels of p-STAT3, TAZ, Beclin1, YAP1, P-gp and LRP in lung cancer cell lines with low expression of YAP1 were significantly decreased (P < 0.05). The expression levels of TAZ, Beclin1, YAP1 and P-gp protein in lung cancer cell lines with high expression of YAP1 were significantly increased (P < 0.05). In the transplanted tumor model of mice, tumor of high-expression Beclin1 group and high-expression YAP1 group were larger than control group, while tumor of low-expression Beclin1 group and low-expression YAP1 group were smaller than control group. Conclusion Xiaoyan Decoction directly affects the expression of YAP1 through Beclin1, and Xiaoyan Decoction can affect the expression of Beclin1 through YAP1-TAZ pathway. Xiaoyan Decoction may reverse the cisplatin resistance of lung cancer by intervening Beclin1-YAP1 loop.

R285.5

国家自然科学基金资助项目（81904151）