自噬相关蛋白4A对胃癌细胞干性及上皮间质转化的影响

doi:10.3969/j.issn.1006-5725.2019.16.004

实用医学杂志 ›› 2019, Vol. 35 ›› Issue (16): 2531-2536.doi: 10.3969/j.issn.1006-5725.2019.16.004

自噬相关蛋白4A对胃癌细胞干性及上皮间质转化的影响

蒋海兵¹, 王伟², 盘箐³, 李国庆¹

南华大学附属第二医院 1消化内科, 2肿瘤科（湖南衡阳 421001）;
3永州职业技术学院病原生物与免疫学教研室（湖南永州 425100）

收稿日期:2019-03-13 出版日期:2019-08-27 发布日期:2019-08-27
通讯作者: 王伟 E-mail：wangwei5918@126.com
基金资助:
湖南省卫生计生委2017 年度第二批科研计划课题项目（编号：A2017014）

Effects of ATG4A on stem and EMT of gastric cancer cells

JIANG Haibing^*, WANG Wei, PAN Qing, LI Guoqing

*Department of Gastroenterology, Second Affiliated Hospital of Nanhua University, Hengyang 421001, China

Received:2019-03-13 Online:2019-08-27 Published:2019-08-27
Contact: WANG Wei E-mail： wangwei5918@126.com

摘要/Abstract

摘要： 目的研究自噬相关蛋白A4（autophagy associated protein 4A,ATG4A）对胃癌细胞干性、上皮间质转化（epithelial mesenchymal transition,EMT）的影响。方法将胃癌细胞株分为：对照组（干扰及过表达ATG4A空白载体组）、干扰组（Sh-ATG4A-1、2、3）、过表达组（ATG4A-OE）。采用划痕试验检测胃癌细胞迁移能力;Transwell试验检测胃癌细胞侵袭能力;通过成球培养法研究ATG4A对胃癌细胞成球能力影响;通过Western Blot检测胃癌干性标志物SOX-2、OCT-4、Bmi-1表达变化,以及EMT标志物E-cadherin、N-cadherin、vimentin表达变化。结果干扰ATG4A表达后,胃癌细胞MGC-803迁移能力下降,而过表达ATG4A后,胃癌细胞SGC-7901迁移能力上升;Transwell侵袭实验发现,干扰ATG4A 表达后,胃癌细胞 MGC-803 侵袭能力下将,而过表达 ATG4A 后,胃癌细胞 SGC-7901侵袭能力上升;干扰 ATG4A 表达后,胃癌细胞 MGC-803 成球能力下将,而过表达ATG4A后,胃癌细胞 SGC-7901 成球能力上升;Western Blot检测显示,干扰 ATG4A 分子表达后,胃癌细胞中 E-cadherin蛋白表达增加,N- cadherin,vimentin蛋白表达下降,而干扰 ATG4A 表达后,SOX-2、OCT-4、Bmi-1蛋白表达水平均下降。结论 ATG4A表达与侵袭转移能力、上皮间质转化能力以及胃癌细胞干性呈正相关。

关键词: 胃癌细胞, 上皮间质转化, 自噬相关蛋白4A

Abstract: Objective To study the effects of autophagy associated protein 4A（ATG4A） on the dry and epithelial-mesenchymal transition （EMT） of gastric cancer cells. Methods Gastric cancer cell lines were divided into groups： Control group （interference and overexpression ATG4A blank vector group）, interference group （Sh-ATG4A-1, 2, 3）, overexpression group （ATG4A-OE）. The migration ability of gastric cancer cells was detected by scratch assay. The invasive ability of gastric cancer cells was detected in transwell system. The expression of SOX-2, OCT-4, Bmi-1, E-cadherin, N-cadherin and vimentin were detected by Western blot. Results cratch assay showed that the migration ability of gastric cancer cell MGC-803 was decreased after interfering with ATG4A expression, while that of SGC-7901 was increased after overexpressing ATG4A. Transwell invasion assay showed that the invasive ability of gastric cancer cells MGC-803 was reduced by interfering with ATG4A expression, while that of SGC-7901 was increased by overexpressing ATG4A. After interfering with ATG4A expression, MGC-803 cells spheroids were decreased, while SGC-7901 cells spheroids were increased after over-expression of ATG4A. Western blot Results showed that the expression of E-cadherin increased and the expression of N-cadherin and vimentin were decreased in gastric cancer cells after interfering with ATG4A expression, while the expression levels of SOX-2, OCT-4 and Bmi-1 were decreased after interfering with ATG4A expression. Conclusion The expression of ATG4A is positively correlated with invasion and metastasis, epithelial-mesenchymal transformation and gastric cancer cell trunks.

Key words: gastric cancer cells, epithelial mesenchymal transformation, autophagy associated protein 4A

蒋海兵, 王伟, 盘箐, 李国庆. 自噬相关蛋白4A对胃癌细胞干性及上皮间质转化的影响[J]. 实用医学杂志, 2019, 35(16): 2531-2536.

JIANG Haibing, WANG Wei, PAN Qing, LI Guoqing. Effects of ATG4A on stem and EMT of gastric cancer cells[J]. The Journal of Practical Medicine, 2019, 35(16): 2531-2536.

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自噬相关蛋白4A对胃癌细胞干性及上皮间质转化的影响

Effects of ATG4A on stem and EMT of gastric cancer cells

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