蛋白激酶R样内质网激酶参与维生素E琥珀酸酯激活人胃癌细胞内质网应激和自噬的调控  

Protein kinase R-like endoplasmic reticulum kinase is involved in regula⁃tion of endoplasmic reticulum stress and autophagy induced by vitamin E succinate in human gastric cancer cells

作　　者：卓翠竹 闻新奕 赵海霞 段宵阳 侯丽颖 ZHUO Cuizhu;WEN Xinyi;ZHAO Haixia;DUAN Xiaoyang;HOU Liying(School of Public Health,North China University of Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学公共卫生学院,河北唐山063210

出　　处：《中国病理生理杂志》2023年第5期788-793,共6页Chinese Journal of Pathophysiology

基　　金：河北省自然科学基金资助项目(No.H2019209453);华北理工大学研究生创新项目

摘　　要：目的:研究蛋白激酶R样内质网激酶(PERK)在维生素E琥珀酸酯(VES)激活人胃癌细胞发生内质网应激(ERS)与自噬中的影响。方法:体外培养人胃癌细胞系MKN28和MKN45,通过CCK-8法绘制细胞生长曲线,根据生长曲线确定用药剂量。不同剂量的VES处理细胞24 h后,qRT-PCR检测葡萄糖调节蛋白78(GRP78)和beclin-1的mRNA表达,Western blot检测GRP78、微管相关蛋白1轻链3(LC3)、beclin-1及PERK的蛋白表达。用2μmol/L GSK2606414(GSK)抑制PERK,设置对照组、GSK组、VES组和VES+GSK组,Western blot检测PERK、GRP78、LC3和beclin-1的蛋白表达。结果:CCK-8实验结果显示,VES对两种人胃癌细胞系的活力均有显著抑制作用(P<0.01)。qRT-PCR结果显示,两种细胞中beclin-1和GRP78的mRNA表达均随着VES剂量的增加而不断升高(均P<0.05)。Western blot结果显示,与对照组相比,VES均可以显著上调两种细胞中GRP78、PERK、LC3和be⁃clin-1的表达(均P<0.05);抑制PERK后,两种细胞中p-PERK/PERK的比值显著下降(P<0.01),相较VES组,VES+GSK组GPR78、LC3和beclin-1的蛋白表达均显著降低(均P<0.05)。结论:VES激活人胃癌细胞发生ERS和自噬的同时上调PERK的表达,而PERK参与了VES对ERS和自噬的调节。AIM:To investigate the effect of protein kinase R-like endoplasmic reticulum kinase(PERK)on the endoplasmic reticulum stress(ERS)and autophagy induced by vitamin E succinate(VES)in human gastric cancer cells.METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro,and the cell viability was detected by CCK-8 assay.The cells were treated with VES at different doses for 24 h.The mRNA expression levels of glu⁃cose-regulated protein 78(GRP78)and beclin-1 were detected by qRT-PCR,while the protein levels of GRP78,microtu⁃bule-associated protein 1 light chain 3(LC3),beclin-1 and PERK were detected by Western blot.After inhibiting PERK with 2μmol/L GSK2606414(GSK),the cells were divided into control group,GSK group,VES group and VES+GSK group,and Western blot was used to detect the expression levels of PERK,GRP78,LC3 and beclin-1.RESULTS:The results of CCK-8 assay illustrated that VES significantly inhibited the viability of the two human gastric cancer cell lines(P<0.01).The qRT-PCR results showed that beclin-1 and GRP78 mRNA expression levels in both cell lines were increased in a dose-dependent manner in response to VES(P<0.05).The Western blot results revealed that VES significantly in⁃creased the expression levels of GRP78,PERK,LC3 and beclin-1 compared with control group(P<0.05).The Western blot results also showed that the ratio of p-PERK/PERK in the two cell lines was decreased drastically in VES+GSK group(P<0.05),and the expression levels of GPR78,LC3 and beclin-1 proteins in VES+GSK group were significantly de⁃creased compared with VES group(P<0.05).CONCLUSION:Vitamin E succinate activates ERS and autophagy in hu⁃man gastric cancer cells,and PERK is involved in the regulation of ERS and autophagy induced by VES.

关 键 词：胃癌 维生素E琥珀酸酯 内质网应激 自噬 蛋白激酶R样内质网激酶 

分 类 号：R735.2[医药卫生—肿瘤] R363.2[医药卫生—临床医学]