自噬在CaSR介导的非小细胞肺癌细胞系A549迁移中的作用  

Autophagy in CaSR-mediated migration of non-small-cell carcinoma cell line A549

作　　者：刘佳龙 李春临 李光伟[1] LIU Jia-long;LI Chun-lin;LI Guang-wei(Department of Pathophysiology, School of Basic Medicine, Qiqihar Medical University, Qiqihar 161006, China)

机构地区：[1]齐齐哈尔医学院基础医学院病理生理学教研室,黑龙江齐齐哈尔161006

出　　处：《基础医学与临床》2020年第11期1479-1483,共5页Basic and Clinical Medicine

基　　金：大学生创新创业项目(201811230021);齐齐哈尔市科技攻关计划(SFGG-201627);齐齐哈尔医学院博士基金(QY2016B-16)。

摘　　要：目的探究自噬在低氧活化的钙敏感受体(CaSR)介导的非小细胞肺癌细胞系A549迁移作用。方法将对数期的A549细胞随机分为对照组(N)、低氧组(H)、低氧CaSR激动剂组(H+Gd)、低氧CaSR抑制剂组(H+NPS)、自噬通路抑制剂组(H+Gd+3-MA)。Western blot检测细胞中CaSR、自噬蛋白(Beclin-1)的表达;细胞划痕实验检测细胞的迁移;Transwell小室法检测细胞的侵袭能力。结果低氧导致A549细胞的CaSR表达增加(P<0.05)、自噬效应蛋白Beclin-1表达量下降(P<0.05)、单位时间细胞愈合百分比上升(P<0.05)、细胞侵袭透膜数量增加(P<0.05)。氯化钆(GdCl3)能够放大低氧的作用,NPS2390可以减弱低氧的作用,自噬抑制剂3-MA在低氧和GdCl3基础上进一步增加A549细胞转移相关指标的表达、而自噬相关指标恰恰相反。结论CaSR通过下调自噬促进A549细胞迁移及侵袭。Objective To investigate the role of autophagy in hypoxia-activated CaSR(calcium-sensing receptor,CaSR)in mediating migration of A549 cells.Methods A549 cells in the logarithmic growth phase were randomly divided into control group(N),hypoxia group(H),hypoxia agonist group(H+Gd),hypoxia inhibitor group(H+NPS),and autophagy pathway inhibitor group(H+Gd+3-MA).Western blot was used to detect the expression of CaSR and autophagy protein(Beclin-1)in the cells;Cell scratch experiment was used to detect migration of A549 cells;Transwell assay was applied to evaluate the invasion of A549 cells.Results Hypoxia resulted in increased CaSR expression in A549 cells(P<0.05),a decreased expression of autophagic effector protein Beclin-1(P<0.05),increased the percentage of cell healing per unit time(P<0.05),and the number of cell invasion-permeable membranes(P<0.05).GdCl3 strengthened the effect of hypoxia,while NPS2390 did weaken the effect of hypoxia.On the basis of hypoxia and GdCl3 conditions,3-MA further increased the expression of metastasis related indicators of A549 cells,but not the autophagy related indicators.Conclusions CaSR activation promotes migration and invasion of A549 cells by down-regulating autophagy.

关 键 词：A549细胞 低氧 自噬 迁移和侵袭 

分 类 号：R363.21[医药卫生—病理学] R730.131[医药卫生—基础医学]