组蛋白去乙酰化酶5在非小细胞肺癌迁移中的作用研究  

Effects of histone deacetylase 5 gene on migration of human non-small cell lung cancer

作　　者：洪伟俊[1] 宁允叶[1] 胥武剑[1] 邵艳[1] 李强[1] 

机构地区：[1]第二军医大学长海医院呼吸内科,上海200433

出　　处：《中国癌症杂志》2011年第4期277-282,共6页China Oncology

摘　　要：背景与目的:组蛋白乙酰化状态受组蛋白去乙酰化酶(histone deacetylases,HDACs)和组蛋白乙酰化酶(histone acetyltransferases,HATs)调节,进而调控基因转录。HDACs和HATs失衡是肿瘤发生发展的重要分子机制。组蛋白去乙酰化酶5(histone deacetylase 5,HDAC5)在多数肿瘤中表达降低,然而其在非小细胞肺癌细胞迁移中的作用尚未见报道。本研究旨在检测HDAC5基因在非小细胞肺癌中的差异性表达,研究HDAC5基因过表达对肺腺癌NCI-H1299细胞迁移的影响。方法:采用real time RT-PCR、Western blot检测28例非小细胞肺癌手术标本中HDAC5 mRNA和蛋白表达;将野生型HDAC5(HDAC5wt)基因转染到NCI-H1299细胞中,将细胞分为转染HDAC5wt基因的实验组(HDAC5wt)、未转染任何质粒的正常对照组(control)、转染空质粒的阴性对照组(vector)。通过MTT方法比较3组细胞的生长情况;利用划痕、Transwell方法检测HDAC5基因过表达对细胞迁移能力的影响。结果:对照组相比,约71.4%(20/28)非小细胞肺癌患者手术标本中HDAC5 mRNA和蛋白表达明显降低;MTT结果显示在第2～5天,实验组D490值显著性低于正常对照组及阴性对照组,而在24 h时,3组细胞增殖差异无统计学意义;Transwell迁移实验结果显示实验组跨膜细胞的数目为158.3±19.4,较正常对照组(247.0±12.1)及阴性对照组(236.0±14.9)均显著减少(P<0.05);划痕实验结果显示HDAC5基因过表达后,H1299细胞划痕愈合程度明显降低。结论:HDAC5基因在非小细胞肺癌中低表达;体外过表达HDAC5基因可抑制NCI-H1299细胞迁移。Background and purpose:The acetylation status of histone which is subjected to histone deacetylases(HDACs) and histone acetyltransferases(HATs) regulates the structure of chromatin as well as the expression of genes.The imbalance of HDACs and HATs may contribute to tumorigenesis and tumor development.Histone deacetylase 5(HDAC5) shows low expression in many kinds of tumors.It remains unclear whether or not the HDAC5 gene affects the migration of non-small cell lung cancer(NSCLC).This study aimed to detect the expression of the HDAC5 gene in NSCLC and para-cancerous tissues,and evaluate the effect of over-expressed HDAC5 on cell migration in lung adenocarcinoma cell NCI-H1299 in vitro.Methods:The levels of HDAC5 protein in 28 NSCLC and para-cancerous tissues were detected by real time RT-PCR and Western blot techniques.The effect of HDAC5 over-expression on H1299 cell migration was examined in vitro.The cells were divided into 3 groups:the HDAC5 over-expression group,which was transfected with HDAC5wt plasmid,the control group,which did not transfect any vector,and the negative control group,which was transfected with an empty vector.MTT assay was used to evaluate cell proliferation.Cell migration was determined through scratch wound healing and transwell assays.Results:The expression of HDAC5 was lower in NSCLC than in the para-cancerous tissues.MTT assay showed that D490 from the HDAC5 over-expression group was significantly lower than that in the other two groups on days 2,3,4 and 5,whereas there was no statistical difference among the 3 groups at 24 h.The number of trans-membrane migration cells in the HDAC5 overexpression group(158.3±19.4) was lower than that in the control group(247.0±12.1) as well as in the negative group(236.0±14.9),as determined by transwell assay(P<0.05).The scratch wound healing assay showed a slower wound closure when HDAC5 was over-expression in H1299 cells.Conclusion:HDAC5 expression was lower in NSCLC tissues than in the para-cancerous tissues,and over-expression of the HDAC5 gen

关 键 词：非小细胞肺癌 组蛋白去乙酰化酶5 增殖 迁移 

分 类 号：R73[医药卫生—肿瘤]