摘要
目的研究维生素E琥珀酸酯(vitamin E succinate,VES)诱导人胃癌细胞自噬过程中肌醇激酶1(inositol-requiring enzyme 1,IRE1)发挥的作用。方法体外培养人胃癌SGC-7901细胞,设置溶剂对照组(0.1%无水乙醇)、不同剂量(5、10、15、20μg/mL)VES组、4μ8C组(4μ8C为IRE1抑制剂)和VES+4μ8C组。Western blot检测对照组、不同剂量VES组内质网应激相关分子葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)、C/EBP同源蛋白(C/EBP homologous protein,CHOP),自噬标志物微管相关蛋白1轻链3(microtubule associated protein1 light chain 3,LC3)、Beclin-1,未折叠蛋白反应分支通路肌醇激酶1(inositol-requiring enzyme 1,IRE1)、X-盒结合蛋白1(X box-binding protein 1,XBP1)、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、p-JNK的蛋白表达水平;用4μ8C抑制IRE1,Western blot检测IRE1、XBP1、JNK、p-JNK、GRP78和CHOP的表达变化,同时检测LC3和Beclin-1的变化情况。结果20μg/mL VES组GRP78的表达(1.16±0.06)、CHOP的表达(1.36±0.11)均较溶剂对照组GRP78(0.36±0.10)、CHOP(0.48±0.05)明显提高(P<0.001);20μg/mL VES组Beclin-1的表达(1.09±0.20)、LC3-Ⅱ/LC3-Ⅰ的表达(1.29±0.03)较溶剂对照组Beclin-1(0.27±0.07)、LC3-Ⅱ/LC3-Ⅰ(0.43±0.06)明显增加(P<0.001);20μg/mL VES组IRE1(1.07±0.20)、XBP1(1.33±0.07)、p-JNK/JNK(1.19±0.31)的表达水平与溶剂对照组相比明显上升(P<0.01);抑制IRE1后:VES+4μ8C组的IRE1(0.63±0.27)、XBP1(0.74±0.09)、p-JNK/JNK(0.35±0.04)、GRP78(0.66±0.02)、CHOP(0.51±0.02)、LC3-Ⅱ/LC3-Ⅰ(0.72±0.01)、Beclin-1(0.70±0.15)的表达水平均较VES组显著降低(P<0.05)。结论VES可通过上调IRE1通路参与对人胃癌细胞自噬的调控。
OBJECTIVE To investigate the role of inositol-requiring enzyme 1(IRE1)in autophagy of human gastric cancer cells induced by vitamin E succinate(VES).METHODS Human gastric cancer SGC-7901 cells were cultured in vitro and divided into solvent control group(0.1%ethanol absolute),different doses(5,10,15 and 20μg/mL)VES group,4μ8C group,and VES+4μ8C group.The endoplasmic reticulum stress-related molecules glucose regulated protein 78(GRP78)and C/EBP homologous protein(CHOP),autophagy marker microtubule associated Protein1 light chain 3(LC3),Beclin-1,unfolded protein response branching pathway Inositol-requiring enzyme 1(IRE1),X box-binding protein 1(XBP1),c-Jun n-terminal kinase(JNK)and p-JNK were detected by Western blot in the solvent control group and different doses of VES group.IRE1 was inhibited by 4μ8C.The expressions of IRE1,XBP1,JNK,p-JNK,GRP78 and CHOP were detected by Western blot,and the expressions of LC3 and Beclin-1 were detected.RESULTS The expression of GRP78(1.16±0.06)and CHOP(1.36±0.11)in 20μg/mL VES group were significantly higher than those in solvent control group GRP78(0.36±0.10)and CHOP(0.48±0.05)(P<0.001).The expression of Beclin-1(1.09±0.20)and LC3-Ⅱ/LC3-Ⅰ(1.29±0.03)in 20μg/mL VES group were significantly higher than those in solvent control group(0.27±0.07)and LC3-Ⅱ/LC3-Ⅰ(0.43±0.06)(P<0.001).The expression levels of IRE1(1.07±0.20),XBP1(1.33±0.07)and p-JNK/JNK(1.19±0.31)in 20μg/mL VES group were significantly higher than those in the solvent control group(P<0.01).After IRE1 is inhibited:The expression level of IRE1(0.63±0.27),XBP1(0.74±0.09),p-JNK/JNK(0.35±0.04),GRP78(0.66±0.02),CHOP(0.51±0.02),LC3-Ⅱ/LC3-Ⅰ(0.72±0.01),Beclin-1(0.70±0.15)was significantly lower than that of VES group(P<0.05).CONCLUSION VES may participate in the regulation of autophagy in gastric cancer cells by upregulating IRE1 pathway.
作者
冯雪
李碧彤
赵海霞
段宵阳
王弈丹
侯丽颖
Feng Xue;Li Bitong;Zhao Haixia;Duan Xiaoyang;Wang Yidan;Hou Liying(School of Public Health,North China University of Technology,Tangshan 063210,China;School of Science,North China University of Technology,Tangshan 063210,China)
出处
《卫生研究》
CAS
CSCD
北大核心
2023年第1期109-114,共6页
Journal of Hygiene Research
基金
河北省自然科学基金(No.H2019209453)
华北理工大学大学生创新创业训练计划(No.X2022199)
华北理工大学大学生创新创业训练计划(No.X2020130)。
关键词
胃癌
维生素E琥珀酸酯
内质网应激
自噬
肌醇激酶1
gastric cancer
vitamin E succinate
endoplasmic reticulum stress
autophagy
inositol-requiring enzyme 1
