下调Nur77抑制PI3K/AKT信号通路促进肺腺癌细胞凋亡和自噬OA北大核心CSTPCD
Downregulation of Nur77 accelerates lung adenocarcinoma cells apoptosis and autophagy through the inhibition of the PI3K/AKT pathway
目的:探讨Nur77对肺腺癌细胞凋亡和自噬的影响及其调控机制.方法:收集右江民族医学院附属西南医院(百色市人民医院)经病理确诊的肺腺癌及癌旁组织5例,免疫荧光法检测Nur77的表达情况;生物信息学分析Nur77相关基因通路;体外培养人支气管上皮细胞BEAS-2B和人肺癌A549两株细胞系,分别记为BEAS-2B组和A549 组.RT-qPCR检测两组细胞Nur77 mRNA的表达水平,Western blot检测Nur77、LC3、PI3K、p-PI3K、AKT、p-AKT的表达情况;免疫荧光检测Nur77的表达水平及亚细胞定位;将A549细胞分为si-NC组、si-Nur77组及Con组,si-NC组及si-Nur77组分别转染si-NC、si-Nur77,Con组未进行转染处理.RT-qPCR检测si-NC、si-Nur77组细胞Nur77 mRNA表达水平;Western blot检测Nur77、LC3、PI3K、p-PI3K、AKT、p-AKT、caspase-3、Bax、Bcl-2蛋白表达水平;流式细胞术检测细胞凋亡率.结果:肺腺癌组织Nur77蛋白表达水平高于癌旁组织;生物信息学分析显示Nur77与PI3K/AKT信号通路存在相关性;A549细胞Nur77 mRNA和蛋白表达水平均高于BEAS-2B细胞(P<0.001),A549细胞LC3Ⅱ/LC3Ⅰ比值、p-AKT/AKT比值均高于BEAS-2B细胞(P<0.05),A549细胞p-PI3K/PI3K比值高于BEAS-2B细胞(P<0.01);si-Nur77组Bcl-2表达低于si-NC组(P<0.05),si-Nur77组Caspase-3、Bax、LC3Ⅱ/LC3Ⅰ比值高于si-NC组(P<0.01),而p-AKT/AK、p-PI3K/PI3K在si-Nur77组低于si-NC组(P<0.05),si-Nur77组细胞凋亡率高于si-NC组(P<0.05).结论:下调Nur77可促进肺腺癌细胞自噬和凋亡,其作用机制可能与抑制PI3K/AKT信号通路有关.
Objective:To investigate the role and mechanism of Nur77 in lung adenocarcinoma cell apoptosis and autophagy.Methods:Five cases of lung adenocarcinoma tissues which were confirmed by pathology and para-cancerous tissues were collected from the Affiliated Southwest Hospital of Youjiang Medical University for Nationalities(People's Hospital of Baise).The expres-sion of Nur77 in lung adenocarcinoma and para-cancerous tissues was detected with immunofluorescence.Nur77-associated path-ways were analyzed with bioinformatics analysis.Human bronchial epithelial(BEAS-2B)cells and human lung cancer(A549)cells were cultured in vitro and designated as BEAS-2B and A549 groups,respectively.In both groups,Nur77 mRNA expression was measured with RT-qPCR,the expression of Nur77,LC3,PI3K,p-PI3K,AKT,and p-AKT was assessed with western blotting,and the fluorescence intensity of Nur77 was determined with immunofluorescence.A549 cells were transfected with si-NC or si-Nur77,referred to as si-NC or si-Nur77 groups,respectively,and non-transfected cells serving as the control group(Con).Nur77 expression was analyzed with RT-qPCR,and the expression of Nur77,LC3,PI3K,p-PI3K,AKT,p-AKT,cas-pase-3,Bax,and Bcl-2 was examined using Western blot.Cell apoptosis was assessed by flow cytometry.Results:Lung adenocar-cinoma tissues exhibited higher levels of Nur77 protein expression compared to para-cancerous tissues.Bioinformatics analysis demonstrated a correlation between Nur77 and the PI3K/AKT pathway.The mRNA and protein expression of Nur77(P<0.001)and the ratio of LC3Ⅱ/LC3Ⅰ,p-AKT/AKT(P<0.05),and p-PI3K/PI3K(P<0.01)were higher in the A549 group than in the BEAS-2B group.Compared with the si-NC group,the si-Nur77 group exhibited decreased Bcl-2 expression(P<0.05),in-creased Caspase-3 and Bax expression(P<0.01),and elevated LC3Ⅱ/LC3Ⅰ ratio(P<0.01),along with reduced p-AKT/AKT(P<0.05)and p-PI3K/PI3K(P<0.01)ratios and significantly enhanced apoptosis(P<0.05).Conclusion:Nur77 down-regulation accelerates autophagy and apoptosis in lung cancer cells,which may be achieved by the inhibition of the PI3K/AKT pathway.
莫黎芳;李小玲;韩谊;周娇;覃春艳;蒋玉洁
右江民族医学院研究生院,广西 百色 533000||百色市人民医院重症医学科,广西 百色 533000右江民族医学院研究生院,广西 百色 533000||右江民族医学院附属医院呼吸与危重症科,广西 百色 533000右江民族医学院研究生院,广西 百色 533000百色市人民医院病理科,广西 百色 533000右江民族医学院附属医院呼吸与危重症科,广西 百色 533000
临床医学
Nur77肺腺癌PI3K/AKT信号通路凋亡自噬
Nur77Lung adenocarcinomaPI3K/AKT pathwayAutophagyApoptosis
《海南医学院学报》 2024 (012)
897-904 / 8
This study was supported by the National Natural Science Foundation of China(81860021);the General Project of Guangxi Natural Science Foundation(2021GXNSFAA325003);the Research Basic Ability Improvement Project of Young and Middle-aged Teachers in Guangxi Universities(2021KY0540);the Science Research and Technology Development Plan of Baise(20193117);the Scientific Research of High-level Talents of the Scientific Research and Training of High-Level Talents(Y202011722)国家自然科学基金资助项目(81860021);广西自然科学基金项目(2021GXNSFAA325003);广西高校中青年教师科研基础能力提升项目(2021KY0540);百色市科学研究与技术开发计划(百科 20193117);右江民族医学院附属医院 2020 年度高层次人次科研项目(Y202011722)
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