重楼皂苷Ⅰ基于miR-16-5p调控STAT3信号通路抑制肺癌细胞增殖和迁移

PolyphyllinⅠbased on miR-16-5p regulates STAT3 signaling pathway to inhibit proliferation and migration of lung cancer cells

目的:探讨重楼皂苷Ⅰ(PPⅠ)通过调控miR-16-5p进而影响STAT3信号通路从而抑制肺癌进展的作用机制。方法:通过网络药理学筛选关键靶标和相关途径;CCK-8、细胞划痕实验、Transwell检测肺癌细胞在PPⅠ作用下增殖、迁移、侵袭能力的变化;通过Western blot测定相关蛋白的表达情况;将miR-16-5p mimic、miR-16-5p inhibitor的表达载体转染到肺癌A549细胞中,再次重复上述实验。结果:STAT3信号通路为PPⅠ和肺癌的关键信号通路,PPⅠ影响肺癌细胞增殖和转移,下调Bcl-2、Cyclin D1、MMP-2、Vimentin,以及上调p21、Bax、Cleave-Caspase-3、Cleave-Caspase-9的蛋白表达水平。PPⅠ干预后,肺癌细胞内miR-16-5p表达水平显著上升。过表达miR-16-5p增强了PPⅠ对A549细胞的影响;PPⅠ调控miR-16-5p可使p-STAT3的蛋白表达水平降低。结论:PPⅠ可能通过调控miR-16-5p和STAT3信号通路抑制人肺癌进展,为制定其治疗策略提供新的方案。

OBJECTIVE To investigate the mechanism by which PolyphyllinⅠregulates miR-16-5p and affects the STAT3 signaling pathway to inhibit lung cancer progression.METHODS Screening key targets and related pathways through network pharmacology.CCK-8,cell wound healing and Transwell assay for the effect of polyphyllinⅠon proliferation,migration and invasion of lung cancer cells.Western blot detection of protein expression levels.miR-16-5p mimic,miR-16-5p inhibitor expression vectors were transfected into A549 cells,and repeat the experiment again.RESULTS The STAT3 signaling pathway was a key signaling pathway between polyphyllinⅠand lung cancer.PolyphyllinⅠaffected the proliferation and metastatic ability of lung cancer.PolyphyllinⅠwas able to down-regulate Bcl-2,Cyclin D1,MMP-2,Vimentin and up-regulate the protein expression levels of p21,Bax,Cleave-Caspase-3,Cleave-Caspase-9.A significant increase in miR-16-5p expression was observed in lung cancer cells after polyphyllinⅠtreatment.A549 cells increased polyphyllinⅠuptake when miR-16-5p was overexpressed.Regulation of miR-16-5p by polyphyllinⅠreduced the protein expression level of p-STAT3.CONCLUSION PolyphyllinⅠmay inhibit the development of human lung carcinoma cells by regulating miR-16-5p and STAT3 signaling pathways,providing a new reference for the treatment of lung cancer.