瘦素上调GRP78的表达在肺癌细胞迁移中的作用  

Role of leptin up-regulated GRP78 expression in migration of lung cancer cells

作　　者：吴优其乐 蔺晓菁 吴晶晶 练雪梅 WU Youqile;LIN Xiaojing;WU Jingjing;LIAN Xuemei(Chongqing Key Laboratory for Lipid and Glucose Metabolic Diseases,Key Laboratory of Molecular Biology for Infectious Diseases of Ministry of Education,Chongqing Medical University,Chongqing,400016,China;Departmentof Nutrition and Food Hygiene,School of Public Health and Management,Chongqing Medical University,Chongqing,400016,China;Key Laboratory of Translational Medicine in Major Metabolic Diseases,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)

机构地区：[1]重庆医科大学脂糖代谢性疾病重庆市重点实验室,感染性疾病分子生物学教育部重点实验室,重庆400016 [2]重庆医科大学公共卫生与管理学院营养与食品卫生学教研室,重庆400016 [3]重庆医科大学附属第一医院重大代谢性疾病转化医学重点实验室,重庆400016

出　　处：《第三军医大学学报》2020年第8期799-806,共8页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81973030);重庆市技术创新与应用发展专项(CSTC2019jscx-msxmX0280)。

摘　　要：目的探讨瘦素对肺癌细胞迁移能力的影响及其可能机制。方法利用GEPIA网站,分析来自TCGA数据库的肺癌患者相关数据,寻求瘦素与葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)表达水平和肺癌患者总生存率的相关性;采用划痕实验检测瘦素及GRP78靶向抑制剂HA15对肺癌细胞株A549与H460迁移能力的影响;免疫荧光检测瘦素对肺癌细胞内GRP78表达的影响。最后在PI3K靶向抑制剂Ly294002,mTOR靶向抑制剂雷帕霉素单独处理或与瘦素联合使用后,Western blot检测瘦素对PI3K/mTOR/STAT3信号通路及GRP78表达的影响。结果生存分析显示瘦素或GRP78高表达均与肺癌患者总生存率下降相关(P<0.05);划痕实验显示瘦素可促进肺癌细胞A549与H460迁移(P<0.05),免疫荧光显示瘦素在A549与H460细胞中上调GRP78的表达(P<0.01)。Western blot检测结果显示瘦素可以通过PI3K/mTOR/STAT3信号通路在A549细胞内上调GRP78的表达,而划痕实验显示抑制GRP78可以减弱瘦素的促迁移作用。结论瘦素能够通过PI3K/mTOR/STAT3信号通路上调GRP78的表达来促进肺癌细胞迁移。Objective To investigate the effect of leptin on the migration of lung cancer cell lines A549 and H460 and the potential mechanisms.Methods The GEPIA website was used to analyze the correlation of the expression levels of leptin and glucose-regulated protein-78(GRP78)with overall survival rate in lung cancer patients from TCGA database.Wound healing assay was used to detect the effect of leptin and HA15(GRP78 targeting inhibitor)on the migration of lung cancer A549 and H460 cells;Immunofluorescence assay was used to measure the expression of GRP78 in lung cancer cells.Finally,lung cancer A549 cells were treated with PI3 K targeting inhibitor Ly294002,mTOR targeting inhibitor rapamycin alone or in combination with leptin,and then Western blot assay was used to investigate the impact of leptin on PI3 K/mTOR/STAT3 signaling pathway and GRP78 expression.Results Survival analysis showed that high expression of leptin or GRP78 was closely related to decreased overall survival rate in lung cancer patients(P<0.05).Wound healing test indicated that leptin enhanced the migration of lung cancer A549 and H460 cells(P<0.05).Immunofluorescence assay displayed that leptin upregulated GRP78 expression in A549 and H460 cells(P<0.01).Western blot assay suggested that leptin could upregulate the expression of GRP78 in lung cancer A549 cells through PI3 K/mTOR/STAT3 signaling pathway(P<0.001).Wound healing test showed inhibition of GRP78 attenuated the effect of leptin on promoting migration.Conclusion Leptin promotes the migration of lung cancer cells by up-regulating GRP78 through PI3 K/mTOR/STAT3 signaling pathway.

关 键 词：瘦素 肺癌 葡萄糖调节蛋白78 迁移 

分 类 号：R341[医药卫生—基础医学] K73-37[历史地理—历史学]