摘要
目的探究吡格列酮(PIO)调节腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对肺癌A549细胞顺铂(CDDP)耐药性的影响。方法将肺癌CDDP耐药细胞A549/CDDP随机分为对照组(Control组)、PIO组(10μmol/L PIO)、CDDP组(10μg/mL CDDP)、CDDP+低浓度PIO组(CDDP+L-PIO组,10μg/mL CDDP+5μmol/L PIO)、CDDP+高浓度PIO组(CDDP+H-PIO组,10μg/mL CDDP+10μmol/L PIO)和CDDP+高浓度PIO组+AMPK激活剂AICAR组(CDDP+H-PIO+AICAR组,10μg/mL CDDP+10μmol/L PIO+20 mmol/L AICAR)。CCK-8法检测细胞增殖能力;划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;流式细胞术测定细胞凋亡率;Western Blot检测各组细胞AMPK/mTOR通路蛋白和凋亡相关蛋白Bcl-2、Bax、Caspase-3蛋白表达。结果与Control组相比,PIO组A549/CDDP细胞OD 450值(48 h、72 h)、细胞迁移率、细胞侵袭数目、AMPK磷酸化水平、Bcl-2蛋白表达显著下降(P<0.05),细胞凋亡率、mTOR磷酸化水平、Bax、Caspase-3蛋白表达显著升高(P<0.05)。与CDDP组相比,CDDP+L-PIO组、CDDP+H-PIO组A549细胞OD 450值(48 h、72 h)、细胞迁移率、细胞侵袭数目、AMPK磷酸化水平、Bcl-2蛋白表达显著下降(P<0.05),细胞凋亡率、mTOR磷酸化水平、Bax、Caspase-3蛋白表达显著升高(P<0.05)。AICAR减弱了PIO对肺癌CDDP耐药A549细胞CDDP敏感性的增强作用。结论PIO可能通过抑制AMPK的活化,激活mTOR,增强肺癌A549/CDDP细胞对CDDP的敏感性。
Objective To investigate the effect of pioglitazone(PIO)on cisplatin(CDDP)resistance in A549 cells of lung cancer by regulating adenylate activated protein kinase(AMPK)/mammalian rapamycin target protein(mTOR)signal pathway.Methods Lung cancer CDDP resistant cells A549/CDDP were randomly divided into Control group(Control group),PIO group(10μmol/L PIO),CDDP group(10μg/mL CDDP),CDDP+low-concentration PIO group(CDDP+L-PIO group,10μg/mL CDDP+5μmol/L PIO),CDDP+high concentration PIO group(CDDP+H-PIO group,10μg/mL CDDP+10μmol/L PIO)and CDDP+high concentration PIO+AMPK activator AICAR group(CDDP+H-PIO+AICAR group,10μg/mL CDDP+10μmol/L PIO+20 mmol/L AICAR).CCK-8 method was applied to detect cell proliferation;the cell migration ability was detected by scratch test;Transwell test was applied to detect the invasion ability of cells;the apoptosis rate was measured by flow cytometry;Western blot was applied to detect the expression of AMPK/mTOR pathway protein and apoptosis-related proteins Bcl-2,Bax,Caspase-3 in each group.Results Compared with the control group,the OD 450 value(48 h,72 h),cell migration rate,cell invasion number,AMPK phosphorylation level,and Bcl-2 protein expression of A549 cells in the PIO group were decreased(P<0.05),the apoptosis rate,mTOR phosphorylation level,Bax and Caspase-3 protein expression were increased(P<0.05).Compared with CDDP group,the OD 450 value(48 h,72 h),cell migration rate,cell invasion number,AMPK phosphorylation level and Bcl-2 protein expression of A549 cells in CDDP+L-PIO group and CDDP+H-PIO group were decreased(P<0.05),the apoptosis rate,mTOR phosphorylation level,Bax and Caspase-3 protein expression were increased(P<0.05).AICAR attenuated the enhancement effect of PIO on CDDP sensitivity of lung cancer CDDP-resistant A549 cells.Conclusion PIO may enhance the sensitivity of lung cancer A549/CDDP cells to CDDP by inhibiting AMPK activation and activating mTOR.
作者
张一思
孙静
张凡
李莉蓉
刘秀丽
ZHANG Yisi;SUN Jing;ZHANG Fan;LI Lirong;LIU Xiuli(Clinical Teaching and Research Section of Clinical Medical College of Suzhou Vocational Health College,Suzhou,Jiangsu 215009,China;Department of Oncology,the First Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang 150007,China)
出处
《临床肺科杂志》
2024年第3期411-416,427,共7页
Journal of Clinical Pulmonary Medicine
基金
苏州卫生职业技术学院院级(领雁培育项目)(No.szwzy202108)。
