核纤层蛋白A基因通过MAPK通路调控胃癌细胞HGC27的增殖、迁移及凋亡  

Apoptosis,inhibits proliferation,and migration of gastric cancer cells HGC27 via MAPK pathway by the knockdown of A-type nuclear lamins gene

作　　者：王双 王正蓉 朱丽英 张艺琼 段云峰 黄昶煜东 杨思远 潘卫 WANG Shuang;WANG Zhengrong;ZHU Liying;ZHANG Yiqiong;DUAN Yunfeng;HUANG Changyudong;YANG Siyuan;PAN Wei(Prenatal Diagnosis Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Medical Laboratory Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Public Health,Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Cardiothoracic Surgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学附属医院产前诊断中心,贵州贵阳550025 [2]贵州医科大学医学检验学院,贵州贵阳550025 [3]贵州医科大学公共卫生学院,贵州贵阳550025 [4]贵州医科大学附属医院心外科,贵州贵阳550025

出　　处：《贵州医科大学学报》2024年第6期781-790,802,共11页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82260074,82260165);国家自然科学基金培育项目(I-2020-31)。

摘　　要：目的探究敲低核纤层蛋白A基因(LMNA)对人胃癌细胞HGC27的增殖、迁移及凋亡的影响和可能的作用机制。方法通过生信分析LMNA基因在胃正常组织与胃癌组织中的表达情况;取对数生长期的人胃癌细胞HGC27,根据是否敲低LMNA基因分为敲低LMNA组(si-1、si-2、si-3组)和对照组(si-NC组)共4组,取瞬时转染率最高的si-2组为si-LMNA组;采用CCK-8实验检测细胞活力,Transwell和划痕实验检测细胞的侵袭迁移能力,流式检测细胞凋亡和细胞周期,Western blot检测凋亡相关蛋白Bax、Caspase3、Bcl-2和丝裂源活化蛋白激酶(MAPK)通路中的p38、p-p38、ERK1/2及pERK1/2的表达;在si-LMNA组加入MAPK通路的特异性激活剂Chicoric acid,进一步验证LMNA的表达情况。结果生信分析结果提示LMNA基因在胃癌组织中高表达,LMNA低表达患者生存率更高;与si-NC组相比,si-LMNA组细胞活力下降(P<0.05),凋亡蛋白Bax、Caspase3表达升高、抗凋亡蛋白Bcl-2降低及细胞凋亡率增加(P<0.05),细胞周期阻滞在G0/G1期、S期细胞减少(P<0.05),侵袭与迁移能力降低(P<0.05),p38、ERK1/2磷酸化水平降低(P<0.05);加入激活剂后,LaminA/C表达量增高,p38、ERK1/2磷酸化水平升高,凋亡相关蛋白的表达被逆转,侵袭迁移能力被促进(P<0.05)。结论敲低LMNA基因后能促进胃癌细胞的凋亡,细胞周期被阻滞在G0/G1期,增殖、侵袭及迁移能力被抑制,其作用机制可能与MAPK信号通路密切相关。Objective To explore the effect of knocking down A-type nuclear laminins gene(LMNA)on the proliferation,migration,and apoptosis of human gastric cancer cells HGC27 and its possible mechanism.Methods The expression of LMNA gene in normal gastric tissues and gastric cancer tissues was analyzed by bioinformatics.Human gastric cancer cells HGC27 at logarithmic growth stage were divided into 4 groups,which include LMNA knockdown group(si-1,si-2,si-3)and control group(si-NC group)according to whether LMNA gene was knocked down.The si-2 group with the highest transient transfection rate was used as the si-LMNA group.CCK-8 assay was used to detect cell viability.Transwell and scratch assay were used to detect the invasion and migration ability of cells.Flow cytometry was used to detect apoptosis and cell cycle.Western blot was used to detect the expressions of apoptosis-related proteins Bax,Caspase3,Bcl-2,and p38,p-p38,ERK1/2,and pERK1/2 in mitogen-activated protein kinase(MAPK)pathway.Chicoric acid,a specific activator of MAPK pathway,was added to si-LMNA group to further verify the expression of LMNA.Results The results of bioinformatics analysis suggested that the expression of LMNA gene was high in gastric cancer tissues,and the survival rate of patients with low expression of LMNA was higher.Compared with the si-NC group,the cell viability of si-LMNA group decreased(P<0.05),the expressions of apoptotic proteins Bax and Caspase3 increased,the anti-apoptotic protein Bcl-2 decreased,and the apoptosis rate increased(P<0.05).The cell cycle was blocked in G0/G1 phase,the number of cells in S phase decreased(P<0.05),invasion and migration ability decreased(P<0.05),and the phosphorylation levels of p38 and ERK1/2 decreased(P<0.05).The expression of LaminA/C and phosphorylation levels of p38 and ERK1/2 increased after the addition of the activator.The expression of apoptosis-related proteins was reversed and the invasion and migration ability was promoted(P<0.05).Conclusion Knockdown of LMNA gene can facilitate the apoptosis

关 键 词：核纤层蛋白A基因 细胞 胃肿瘤 丝裂源活化蛋白激酶信号通路 增殖 迁移 

分 类 号：R735.2[医药卫生—肿瘤]