阳和汤加味对肺腺癌细胞的影响及作用机制  

Effect and Mechanism of Modified Yanghe Decoction on Lung Adenocarcinoma Cells

作　　者：孙平一 刘娟[2] 刘思远[2] 王荣 郑心 SUN Pingyi;LIU Juan;LIU Siyuan;WANG Rong;ZHENG Xin(Jining Medical University,Jining 272000,China;Shandong University of Traditional Chinese Medicine,Ji′nan 250000,China;Qingdao Hospital of Traditional Chinese Medicine/Qingdao Haici Hospital,Qingdao 266000,China)

机构地区：[1]济宁医学院,济宁272000 [2]山东中医药大学,济南250000 [3]青岛市中医院/青岛市海慈医疗,青岛266000

出　　处：《世界中医药》2024年第24期3769-3774,共6页World Chinese Medicine

基　　金：国家自然科学基金项目(82374288);山东省自然科学基金项目(ZR2021MH274);青岛市科技惠民示范专项(23-2-8-smjk-4-nsh)。

摘　　要：目的:观察阳和汤加味含药血清对人肺腺癌细胞(A549、H1975)及其miR-1-3p/磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路的影响。方法:构建A549、H1975细胞微小RNA-1-3p(miR-1-3p)过表达及干扰稳转株,给予阳和汤加味含药血清干预,通过肺腺癌细胞增殖及毒性检测实验(CCK-8)、划痕、Transwell实验检测细胞增殖、迁移及侵袭能力;实时定量聚合酶链式反应(RT-qPCR)检测miR-1-3p mRNA表达水平;蛋白质印迹法检测PI3K/AKT/mTOR通路蛋白表达水平。结果:miR-1-3p mRNA表达水平在2种肺腺癌细胞(A549、H1975)中明显低于正常肺泡上皮细胞Beas-2b(P<0.05);与空白血清比较,含药血清组肺腺癌细胞的miR-1-3p表达量增加,细胞增殖、迁移及侵袭能力受到明显抑制,干扰miR-1-3p后逆转了这种抑制作用(P<0.05);与空载组比较,miR-1-3p过表达组细胞磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达量下降(P<0.05);含药血清干预后,2种肺腺癌细胞PI3K/AKT/mTOR信号通路蛋白表达量降低,干扰miR-1-3p可逆转对通路蛋白表达水平的抑制作用(P<0.05)。结论:阳和汤加味含药血清能够抑制肺腺癌细胞A549、H1975的增殖、迁移及侵袭,其作用机制可能与调控miR-1-3p的表达进而抑制PI3K/AKT/mTOR信号通路有关。Objective:To observe the effect of modified Yanghe Decoction(YHD)-medicated serum on human lung adenocarcinoma cells(A549,H1975)and its influence on the miR-1-3p/phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)signaling pathway.Methods:A549 and H1975 cells were established as stable overexpression and interference cell lines of miR-1-3p.The cells were treated with modified YHD-medicated serum.The Cell Counting Kit-8(CCK-8),scratch test,and Transwell assay were used to assess cell proliferation,migration,and invasion of lung adenocarcinoma cells.Real-time quantitative PCR(RT-qPCR)was used to measure miR-1-3p mRNA expression,and Western blot was used to detect the expression levels of proteins in the PI3K/AKT/mTOR pathway.Results:The expression level of miR-1-3p mRNA in A549 and H1975 lung adenocarcinoma cells was significantly lower than that in normal alveolar epithelial cells(Beas-2b)(P<0.05).Compared with the blank serum group,the drug-medicated serum group increased miR-1-3p expression and significantly inhibited cell proliferation,migration,and invasion.This inhibitory effect was reversed after miR-1-3p interference(P<0.05).Compared with the control group,the miR-1-3p overexpression group showed reduced expression of PI3K,AKT,and mTOR proteins(P<0.05).After treatment with drug-medicated serum,the protein expression levels of the PI3K/AKT/mTOR signaling pathway in both cell lines were reduced,and the inhibition of pathway protein expression was reversed by miR-1-3p interference(P<0.05).Conclusion:Modified YHD-medicated serum can inhibit the proliferation,migration,and invasion of lung adenocarcinoma cells(A549,H1975).The mechanism of action may involve the regulation of miR-1-3p expression,which subsequently inhibits the PI3K/AKT/mTOR signaling pathway.

关 键 词：阳和汤加味 经典名方 肺腺癌 药理作用 作用机制 miR-1-3p 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR) 合理运用 

分 类 号：R285.5[医药卫生—中药学]