摘要
目的研究miR-1226对非小细胞肺癌(NSCLC)吉非替尼敏感性的影响,并探讨其作用机制。方法体外培养NSCLC细胞株人肺癌耐吉非替尼细胞株(PC9GR)和人肺腺癌细胞(PC-9);将PC-9组细胞设置为正常组,PC9GR细胞随机分为对照组、miR-1226 mimic组和miR-1226 NC组,并进行转染;利用实时荧光定量PCR(RT-qPCR)法检测转染后各组细胞miR-1226及黏蛋白1(MUC1)mRNA表达情况;采用蛋白印迹分析法检测转染后各组细胞MUC1蛋白表达情况;采用MTT法检测转染后各组细胞对吉非替尼的敏感性;Transwell法检测转染后各组细胞的侵袭能力;采用双荧光素酶报告实验验证miR-1226与MUC1的靶向关系。结果与正常组相比,对照组和miR-1226 NC组PC9GR细胞miR-1226表达水平显著降低(P<0.05),IC50值、MUC1 mRNA及MUC1蛋白相对表达量、细胞侵袭数量显著升高(P<0.05);与对照组和miR-1226 NC组相比,miR-1226 mimic组PC9GR细胞miR-1226表达水平显著升高(P<0.05),IC50值、MUC1 mRNA及MUC1蛋白相对表达量、细胞侵袭数量显著降低(P<0.05);与MUC1-3′UTR-WT+miR-1226 NC组比,MUC1-3′UTR-WT+miR-1226 mimic组荧光素酶活性降低(P<0.05)。结论miR-1226过表达可能通过靶向下调MUC1表达,提高人非小细胞肺癌细胞吉非替尼敏感性。
Objective To study the effect of miR-1226 on the sensitivity of gefitinib in non-small cell lung cancer(NSCLC)and its mechanism.Methods NSCLC cell lines human lung cancer cell line resistant to gefitinib(pc9gr)and human lung adenocarcinoma cell line(PC-9)were cultured in vitro.The cells of the PC-9 group were set as the normal group,and Pc9gr cells were randomly divided into the control group,the miR-1226 mimic group and the miR-1226 NC group,and all of them were transfected.The expressions of miR-1226 and mucin 1(MUC1)mRNA were detected by real-time fluorescence quantification(RT-qPCR).The expression of MUC1 was detected by Western blotting.MTT method was used to detect the sensitivity of each group of cells to gefitinib.Transwell method was used to detect the invasiveness of cells in each group,and the targeting relationship between miR-1226 and MUC1 was verified by double Luciferase Report.Results Compared with the normal group,the expression level of miR-1226 in pc9gr cells in the control group and the miR-1226 NC group was significantly lower(P<0.05),and the IC50,the relative expressions of MUC1 mRNA and MUC1 protein and cell invasion were significantly higher(P<0.05).Compared with the control group and the miR-1226 NC group,the expression level of miR-1226 in pc9gr cells in the miR-1226 mimic group was significantly higher(P<0.05),and the IC50,the relative expressions of MUC1 mRNA and MUC1 protein and cell invasion were significantly lower(P<0.05).Moreover,compared with the MUC1-3′UTR-WT+miR-1226 NC group,the luciferase activity of the MUC1-3′UTR-WT+miR-1226 mimic group was lower(P<0.05).Conclusion The over-expression of miR-1226 may increase the sensitivity of gefitinib in human non-small cell lung cancer cells by targeting down-regulating the MUC1 expression.
作者
冀国发
胡静怡
任徽
JI Guo-fa;HU Jing-yi;REN Hui(Department of Respiratory Medicine,East Hospital of the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an,Shaanxi 710089,China;Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an,Shaanxi 710000,China)
出处
《临床肺科杂志》
2021年第2期240-244,共5页
Journal of Clinical Pulmonary Medicine
基金
陕西省科学技术厅(No.2018SF-071)。
