[关键词]
[摘要]
目的 探讨消岩汤逆转肺癌顺铂(DDP)耐药的可能机制。方法 MTT检测细胞增殖情况,划痕实验检测细胞侵袭能力;siRNA转染细胞获取稳定低表达基因的细胞株;实时荧光定量PCR(qRT-PCR)和Werstern blotting检测mRNA和蛋白表达水平的变化。结果 MTT检测A549/DDP,在72、96 h,消岩汤能够明显抑制细胞增殖,而在48、72、96 h,DDP联合消岩汤能够明显抑制细胞增殖。划痕实验显示与A549/DDP/siRNA-NC比较,A549/DDP/siRNA-Beclin1细胞株迁移距离明显缩短。MTT显示与A549/DDP/siRNA-NC比较,A549/DDP/siRNA-Beclin1细胞株增殖能力减低,Western blotting显示与A549/DDP/siRNA-NC比较,A549/DDP/siRNA-Beclin1细胞P-糖蛋白(P-gp)和肺耐药相关蛋白(LRP)蛋白表达降低,MTT检测A549/DDP/siRNA-Beclin1细胞,在72、96 h,DDP和消岩汤能明显抑制细胞增殖;而在24、48、72、96 h,DDP联合消岩汤明显抑制细胞增殖。结果显示DDP联合消岩汤使A549/DDP/siRNA-Beclin1细胞株中YES关联蛋白1(YAP1)、P-gp和LRP蛋白表达明显降低。结论 消岩汤可能通过影响Beclin 1-YAP1分子通路进而改变肺癌细胞对DDP的敏感性。
[Key word]
[Abstract]
Objective To investigate the possible mechanism of reversal of cisplatin resistance in lung cancer by Xiaoyan Decoction. Methods The migration ability of cancer cells was observed by the scratch test, and the MTT was used to test the mobility of cancer cells. The siRNA transfection was applied to demonstrate the possible molecular mechanisms. The mRNA and protein expression levels were detected by real-time quantitative reverse transcription PCR and Western Blot. Results MTT showed that A549/DDP could significantly inhibit cell proliferation at later points of time. Similarly, cisplatin combined with Xiaoyan Decoction could significantly inhibit cell proliferation at 48, 72 and 96 h. Scratch test showed that the migration distance was shorter in A549/DDP/siRNA-Beclin1 cell line than NC. MTT showed the proliferation of A549/DDP/siRNA-Beclin1 cell line was decreased when compaired with NC. Western blot also showed that the protein expression of P-gp and LRP was decreased. MTT also detected that cisplatin and Xiaoyan Decoction significantly inhibited cell proliferation of A549/DDP/siRNA-Beclin1 cell at later points of time. Cisplatin combined with Xiaoyan Decoction significantly inhibited cell proliferation in the whole course. The results showed that protein expression level of YAP1, P-gp and LRP in A549/DDP/siRNA-Beclin1 cell line was significantly lower than those in NC. Conclusion Xiaoyan Decoction may improve the sensitivity to cisplatin may via Beclin 1-YAP1 pathway.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金资助项目(81904151);天津市教委科研计划项目(2018KJ015)