摘要
目的研究HOXA-AS3通过调控HOXA6/7基因对人肺腺癌细胞增殖凋亡的作用机制。方法人肺腺癌细胞株经培养后随机分为LA组(单纯人肺腺癌细胞株)和EX组(HOXA-AS3转染的人肺腺癌细胞株)。使用TUNEL法、CCK-8法、划痕实验、Western blot法及RT-PCR法分析两组细胞凋亡、增殖、迁移能力、HOXA6/7蛋白表达及HOXA6/7 mRNA表达的差异。结果TUNEL法结果表明LA组细胞凋亡较EX组多且分布明显稀疏(P<0.05);CCK-8检测结果表明EX组细胞的增殖数量明显高于LA组(P<0.05);划痕实验显示,与EX组相比,LA组无细胞区域宽度较大,表明其细胞迁移能力较低;Western blot法对两组细胞内HOXA6/7蛋白的免疫印迹分析显示,EX组的HOXA6/7蛋白表达明显上升(P<0.05);RT-PCR结果显示,EX组的HOXA6/7 mRNA表达明显升高(P<0.05)。结论HOXA-AS3激活HOXA6/7基因的表达,加快人肺腺癌细胞的增殖、迁移,促进疾病发展。深入研究HOXB6/7基因对肺腺癌的作用机制可为控制肺腺癌的发展与转移提供临床帮助。
Objective To study the mechanism of HOXA-AS3 on the proliferation and apoptosis of humanlung adenocarcinoma cells by regulating HOXA6/7 gene.MethodsHuman lung adenocarcinoma cell lines wererandomly divided into LA group(simple human lung adenocarcinoma cell line)and EX group(transfection ofHOXA-AS3 into human lung adenocarcinoma cell line)after culture.TUNEL method,CCK-8,scratch test,Western Blot and RT-PCR were used to analyze the differences in apoptosis,proliferation,migration ability and theexpression of HOXA6/7 protein and HOXA6/7 m RNA between LA group and EX group.ResultsTUNEL methodshowed that the apoptosis of human lung adenocarcinoma cells in LA group was more than that in EX group,and thedistribution was significantly sparse(P<0.05).CCK-8 method indicated that the proliferation of lungadenocarcinoma cells in EX group was significantly higher than that in LA group(P<0.05).The results of scratchtest showed that,compared with EX group,the width of acellular zone was larger in LA group,indicating lowmigration ability.Immuno Blotting analysis of HOXA6/7 protein of lung adenocarcinoma cells in LA group and EXgroup by Western Blot method showed that the expression of HOXA6/7 protein in EX group increased significantly(P<0.05).RT-PCR results showed that the expression of HOXA6/7 m RNA in EX group was significantly higherthan that in LA group(P<0.05).ConclusionHOXA-AS3 accelerates the proliferation and migration of humanlung adenocarcinoma cells and promotes the development of disease by activating the expression of HOXA6/7 gene.Further study of the effect of HOXB6/7 gene on lung adenocarcinoma is helpful for controlling the development andmetastasis of lung adenocarcinoma.
作者
张超
曲晓翰
韩立波
ZHANG Chao;QU Xiaohan;HAN Libo(The First Hospital of China Medical University,Shenyang 110001,China)
出处
《实用医学杂志》
CAS
北大核心
2020年第17期2371-2375,共5页
The Journal of Practical Medicine
基金
辽宁省博士科研启动基金指导计划项目(编号:20170520349)。
