苍术素调节Notch信号对EGFR-TKI耐药的非小细胞肺癌细胞株H1975增殖、凋亡的影响  

Effect of atractylodin on proliferation and apoptosis of EGFR-TKI resistant non-small cell lung cancer cell line H1975 by regulating Notch signal

作　　者：张琳 朱琳 李晨 ZHANG Lin;ZHU Lin;LI Chen(Department of Pharmacy,Yangpu Hospital,Tongji University,Shanghai 200090,China)

机构地区：[1]同济大学附属杨浦医院药学部,上海200090

出　　处：《临床肺科杂志》2023年第10期1487-1493,1510,共8页Journal of Clinical Pulmonary Medicine

摘　　要：目的探讨苍术素对表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)耐药的非小细胞肺癌(NSCLC)细胞株H1975增殖、凋亡的影响以及可能的作用机制。方法以人肺腺癌H1975细胞为亲本细胞,采用低浓度逐步加量诱导法构建EGFR-TKI耐药细胞株H1975/吉非替尼耐药(GR)。H1975/GR细胞分为正常组(不添加任何药物)、苍术素(低、中、高)浓度组(20、40、60μmol/L苍术素)、Notch抑制剂组(7.8 nmol/L Notch抑制剂1),MTT法检测各组H1975/GR细胞增殖活力;流式细胞仪检测各组H1975/GR细胞周期分布和凋亡率;qRT-PCR和Western Blot检测各组H1975/GR细胞中Notch信号通路相关基因和蛋白表达水平。结果吉非替尼抑制H1975/GR细胞的IC_(50)显著高抑制H1975细胞的IC_(50)(P<0.05),表明本研究所构建的EGFR-TKI耐药细胞株H1975/GR可以用于后续实验。药物干预48 h后,与正常组比较,苍术素(低、中、高)浓度组和Notch抑制剂组H1975/GR细胞增殖活力、G0/G1细胞比例以及细胞中Notch1、Notch3、Hes1、HEY1 mRNA和蛋白表达水平降低(P均<0.05),而G2/M细胞比例、凋亡率升高(P均<0.05),且苍术素各浓度组呈浓度依赖性(P均<0.05);与苍术素(低、中)浓度组比较,Notch抑制剂组H1975/GR细胞增殖活力、G0/G1细胞比例以及细胞中Notch1、Notch3、Hes1、HEY1 mRNA和蛋白表达水平降低(P均<0.05),G2/M细胞比例、凋亡率升高(P均<0.05);与苍术素高浓度组比较,Notch抑制剂组H1975/GR细胞G2/M细胞比例降低(P<0.05)。结论苍术素可抑制EGFR-TKI耐药的NSCLC细胞株H1975增殖,并诱导细胞周期阻滞和凋亡,其作用机制可能与抑制Notch信号通路激活有关。Objective To investigate the effect of atractylodin on the proliferation and apoptosis of non small cell lung cancer(NSCLC)cell line H1975 resistant to epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI)and its possible mechanism.Methods Human lung adenocarcinoma cell line H1975 was used as the parent cells,and EGFR-TKI drug resistant cell line H1975/gefitinib(GR)was constructed by low concentration stepwise induction.H1975/GR cells were divided into the normal group(without adding any drugs),the atractylodin(low,medium and high)concentration groups(20,40,60μmol/L atractylodin),and the Notch inhibitor group(7.8 nmol/L Notch inhibitor 1).MTT method was used to detect the proliferation activity of H1975/GR cells in each group.The cell cycle distribution and apoptosis rate of H1975/GR in each group were detected by flow cytometry.The expression levels of Notch signal pathway related genes and proteins in H1975/GR cells were detected by qRT-PCR and Western blot.Results The IC 50 of H1975/GR cells to gefitinib was obviously higher than that of H1975 cells to gefitinib(P<0.05),which indicated that the EGFR-TKI drug-resistant cell line H1975/GR constructed in this study could be used in subsequent experiments.48 hours after drug intervention,the proliferation activity of H1975/GR cells,the proportion of G0/G1 cells,and the expression levels of Notch1,Notch3,Hes1,HEY1 mRNA and protein in cells decreased obviously in the atractylodin(low,medium,high)concentration groups and the Notch inhibitor group while compared with the normal group(P<0.05).The proportion of G2/M cells and apoptosis rate increased(P<0.05).The atractylodin groups were dependent with their concentration(P<0.05).Compared with the atractylodin(low and medium)concentration groups,the proliferation activity of H1975/GR cells,the ratio of G0/G1 cells,and the expression levels of Notch1,Notch3,Hes1,HEY1 mRNA and protein in cells of the Notch inhibitor group decreased(P<0.05),and the proportion of G2/M cells and apoptosis rate increased(P<0.05

关 键 词：苍术素 非小细胞肺癌 表皮生长因子受体酪氨酸激酶抑制剂 耐药 Notch信号通路 

分 类 号：R734.2[医药卫生—肿瘤]