骨桥蛋白介导PI3K/AKT通路促进非小细胞肺癌顺铂耐药  

Promotion of Cisplatin Resistance by Osteopontin through Mediation of PI3K/AKT Pathway in Non-small Cell Lung Cancer

作　　者：刘迪[1] 龙谦[1] 罗猛[1] 胡剑[1] 陈臣[1] 裴登科 Liu Di;Long Qian;Luo Meng(Department of Thoracic Surgery,The People’s Hospital of Guizhou Province,Guiyang 550002,China)

机构地区：[1]贵州省人民医院胸外科,贵阳550002

出　　处：《华中科技大学学报（医学版）》2020年第1期17-23,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：贵州省科技计划项目(No.2016-7183)。

摘　　要：目的探讨骨桥蛋白(OPN)与非小细胞肺癌(NSCLC)顺铂耐药的相关性及其机制。方法选用NSCLC A549细胞系,构建OPN沉默细胞及OPN过表达细胞;肺癌细胞对顺铂的敏感性利用MTT和流式细胞仪分析;采用Western blot检测顺铂处理前后各组细胞PI3K和ERK1/2蛋白表达水平;分别用实时荧光定量PCR和Western blot检测顺铂处理前后各组细胞ERCC1的mRNA和蛋白表达水平。结果成功构建OPN过表达及OPN沉默载体。经顺铂处理后,OPN过表达组的细胞增殖活性明显高于对照组,OPN沉默组的细胞增殖活性明显低于对照组,差异均具有统计学意义(均P<0.05)。OPN过表达组的细胞凋亡明显低于对照组,差异具有统计学意义(P<0.05)。OPN过表达组的PI3K、p-PI3K蛋白表达水平升高,OPN沉默组的PI3K、p-PI3K蛋白表达水平下降,顺铂处理后OPN过表达组的PI3K、p-PI3K蛋白表达水平较OPN沉默组高,而ERK1/2、p-ERK1/2蛋白表达水平则无明显差异。加入ERK抑制剂U0126,OPN过表达组的细胞增殖活性,OPN沉默组的细胞增殖活性与对照组无明显差异。OPN过表达组的细胞凋亡,OPN沉默组的细胞凋亡与对照组无明显差异。加入PI3K抑制剂LY294002后,对照+LY294002+顺铂组增殖率较对照组低,对照+LY294002+顺铂组、shRNA-OPN+LY294002+顺铂组凋亡率较对照组高,差异具有统计学意义(均P<0.05)。OPN过表达组ERCC1的mRNA表达较对照组明显升高,OPN沉默组ERCC1 mRNA的表达与对照组相比明显降低,差异具有统计学意义(均P<0.05)。OPN过表达组ERCC1的蛋白表达高于对照组,OPN沉默组ERCC1的蛋白表达低于对照组。结论 OPN过表达可促进NSCLC对顺铂耐药。OPN促进NSCLC对顺铂耐药的机制可能是通过激活PI3K/AKT信号通路,而与ERK1/2通路活化无明显相关。OPN过表达可上调ERCC1,OPN可能通过PI3K/AKT信号通路上调ERCC1从而促进NSCLC对顺铂的耐药。Objective To investigate the correlation between osteopontin(OPN)and cisplatin resistance as well as the potential mechanism in non-small cell lung cancer(NSCLC).Methods The NSCLC A549 cell line was used to construct OPN-silenced and OPN-overexpressed cells.The sensitivity of lung cancer cells to cisplatin was analyzed by MTT and flow cytometry.The expression levels of PI3 K and ERK1 were determined by Western blotting,while the expression levels of ERCC1 mRNA and protein in the cells of each group were determined by real-time fluorescent quantitative PCR and Western blotting,respectively.Results Vectors for OPN overexpression and OPN silencing were successfully constructed.After cisplatin treatment,as compared with the control group,the cell viability was significantly increased in the OPN overexpression group and significantly reduced in the OPN silencing group(both P<0.05);the apoptosis of the OPN overexpression group was significantly decreased(P<0.05).The expression levels of PI3 K and p-PI3 K proteins were increased in the OPN overexpression group and decreased in the OPN silencing group.Also,the expression levels of PI3 K and p-PI3 K proteins in the OPN overexpression group were higher than those in the OPN silencing group after cisplatin treatment,while the expression levels of ERK1/2 and p-ERK1/2 protein were comparable.After the addition of ERK inhibitor U0126,no significant differences in cell viability and cell apoptosis were observed in the OPN overexpression group and the OPN silencing group as compared with those of the control group.After the addition of LY294002,a PI3 K inhibitor,the proliferation rate of cells in the control group+LY294002 +cisplatin was found to be lower than that of the control group,and the apoptosis rates of the control group+LY294002+cisplatin group and the shRNA-OPN +LY294002+cisplatin group were significantly higher than that of the control group(both P<0.05).Also,as compared with the control group,the expression levels of ERCC1 mRNA and protein were,respectively,signific

关 键 词：非小细胞肺癌 骨桥蛋白 顺铂 耐药 PI3K/AKT ERK1/2 ERCC1 

分 类 号：R734.2[医药卫生—肿瘤]