AGR2对毒胡萝卜素作用下人肺癌NCI-H460细胞增殖及凋亡的影响  

Effects of AGR2 on proliferation and apoptosis of NCI-H460 cells induced by thapsigargin

作　　者：林志国 刘佳 杜娟 刘振艳 彭瑶 皮秀杰 王玉凤 战文娜 李德海 刘颖 邓志会 LIN Zhi-guo;LIU Jia;DU Juan;LIU Zhen-yan;PENG Yao;PI Xiu-jie;WANG Yu-feng;ZHAN Wen-na;LI De-hai;LIU Ying;DENG Zhi-hui(Respiratory Department,The First Affiliated Hospital,Qiqihar Medical University,Qiqihar 161006,China;Pharmacy Department,The Second Affiliated Hospital,Qiqihar Medical University,Qiqihar 161006,China;Institute of Medicine and Pharmacy,Qiqihar Medical University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔医学院附属第一医院呼吸内科,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院附属第二医院药剂科,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔161006

出　　处：《中国病理生理杂志》2018年第9期1690-1695,共6页Chinese Journal of Pathophysiology

基　　金：齐齐哈尔市科技局基金资助项目(No.SFGG-201504)

摘　　要：目的:观察anterior gradient 2(AGR2)基因在内质网应激诱导剂毒胡萝卜素作用下对人肺癌NCIH460细胞增殖能力及凋亡的影响。方法:通过慢病毒介导的shRNA建立AGR2基因沉默的稳定细胞系;采用集落形成实验检测AGR2在毒胡萝卜素作用下对细胞增殖能力的影响;流式细胞术分析在毒胡萝卜素作用下AGR2对细胞凋亡的影响。结果:成功构建AGR2基因沉默的稳定细胞系,Western blot结果显示AGR2的蛋白表达水平显著降低(P<0.01);在未经药物处理条件下,AGR2基因缺失对NCI-H460细胞集落形成率及凋亡无显著影响;毒胡萝卜素作用24 h后,AGR2基因沉默的细胞集落形成率远低于对照组(P<0.05);流式细胞术检测结果显示AGR2基因缺失提高了毒胡萝卜素诱导的细胞凋亡率(P<0.05)。结论:AGR2基因缺失不影响正常条件下NCI-H460细胞的存活及凋亡能力,但可使毒胡萝卜素作用下细胞的增殖能力显著下降,细胞凋亡率显著提高。本研究揭示了AGR2在内质网应激状态下促进肺癌细胞增殖和减轻细胞凋亡的作用,为进一步探讨AGR2在肺癌发生发展及耐药性中的作用提供了理论依据。AIM:To observe the effects of anterior gradient 2(AGR2)gene on the proliferation and apoptosis of NCI-H460 cells induced by thapsigargin,an endoplasmic reticulum(ER)stress inducer.METHODS:A stable cell line with knockdown of endogenously expressed AGR2 mediated by lentiviral shRNA was established.The proliferation abi-lity of stable NCI-H460 cells in the presence of thapsigargin was measured by colony formation assay,while the effect of AGR2 gene silencing on apoptosis in the presence of thapsigargin was analyzed by flow cytometry.RESULTS:Lentivirus-mediated AGR2 knockdown stable NCI-H460 cells was successfully constructed and the expression of AGR2 was markedly diminished as revealed by Western blot(P<0.01).Knockdown of AGR2 didn’t significantly affect the colony formation rate and apoptosis of NCI-H460 cells without thapsigargin treatment.Colony formation rate of the cells with AGR2 gene knockdown was far lower than that of the control cells after 24 h of thapsigargin treatment(P<0.05).The results of flow cytometry showed that knockdown of AGR2 significantly increased the apoptosis of NCI-H460 cells induced by thapsigargin.CONCLUSION:Knockdown of AGR2 doesn’t remarkably affect the proliferation and apoptosis of NCI-H460 cells in the absence of thapsigargin,but significantly decreases cell viability and increases apoptosis after thapsigargin treatment.This study reveals that AGR2 might promote the proliferation and reduce the apoptosis of lung cancer cells under ER stress,and lays the foundation for further study of the AGR2 role in initiation,development and drug resistance of lung cancer.

关 键 词：AGR2基因 内质网应激 毒胡萝卜素 细胞凋亡 肺癌 

分 类 号：R734.2[医药卫生—肿瘤] R329.25[医药卫生—临床医学]