自噬在CaSR介导的非小细胞肺癌细胞系A549迁移中的作用

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (11): 1479-1483.

自噬在CaSR介导的非小细胞肺癌细胞系A549迁移中的作用

刘佳龙, 李春临, 李光伟^*

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

收稿日期:2020-03-22 修回日期:2020-06-29 出版日期:2020-11-05 发布日期:2020-10-30
通讯作者: ^* 1102044987@qq.com
基金资助:
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)

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

LIU Jia-long, LI Chun-lin, LI Guang-wei^*

Department of Pathophysiology, School of Basic Medicine, Qiqihar Medical University, Qiqihar 161006, China

Received:2020-03-22 Revised:2020-06-29 Online:2020-11-05 Published:2020-10-30
Contact: ^* 1102044987@qq.com

摘要/Abstract

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。

关键词: A549细胞, 低氧, 自噬, 迁移和侵袭

Abstract: 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.

Key words: A549 cell, hypoxia, autophagy, migration and invasion

中图分类号:

刘佳龙, 李春临, 李光伟. 自噬在CaSR介导的非小细胞肺癌细胞系A549迁移中的作用[J]. 基础医学与临床, 2020, 40(11): 1479-1483.

LIU Jia-long, LI Chun-lin, LI Guang-wei. Autophagy in CaSR-mediated migration of non-small-cell carcinoma cell line A549[J]. Basic & Clinical Medicine, 2020, 40(11): 1479-1483.

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