过表达Oct4B1诱导结直肠癌SW480细胞发生上皮间质转化  

Overexpression of Oct4B1 induces epithelial mesenchymal transition in colorectal cancer SW480 cells

作　　者：陈奕霖[1] 文坤明[1] 胡水清[1] 陈正权[1] 曾庆良[1] CHEN Yi-Lin WEN Kun-Ming HU Shui-Qing CHEN Zheng-Quan ZENG Qing-Liang.(Department of Gastrointestinal Surgery, the Affiliated Hospital of Zunyi Medical College, Zunyi 563000, Chin)

机构地区：[1]遵义医学院附属医院胃肠外科,遵义563000

出　　处：《中国免疫学杂志》2017年第5期656-659,664,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81260369);贵州省科技厅科学技术基金[黔科合(J)字(2012)2365]资助

摘　　要：目的:探讨Oct4B1基因过表达是否诱导人结直肠癌SW480细胞发生上皮间质转化(EMT)及其可能的机制。方法:用带G418抗性的Oct4B1基因过表达质粒及阴性对照质粒转染人结直肠癌SW480细胞株,分别称为实验组(SW480-Oct4B1)及对照组(SW480-NC),转染成功后用G418筛选建立稳定转染的细胞株,对两种稳定转染细胞进行如下检测:(1)RTq PCR检测Oct4B1的mRNA水平;(2)划痕实验和Transwell小室实验检测迁移和侵袭能力;(3)Western blot检测EMT相关标记物E-cadherin、N-cadherin及Vimentin蛋白表达;(4)RT-q PCR和Western blot检测EMT转录因子Twist的mRNA及蛋白表达。结果:稳定转染细胞株建立后,实验组与对照组比较:Oct4B1基因表达水平明显升高(P<0.01);细胞迁移明显增强(P<0.01);细胞侵袭能力明显提高(P<0.01);上皮标记物E-cadherin蛋白表达量明显下降(P<0.01);而间质标记物N-cadherin及Vimentin蛋白表达量明显上调(P<0.01);Twist的mRNA及蛋白表达量均明显升高(P<0.01)。结论:过表达Oct4B1基因可诱导人结直肠癌SW480细胞发生EMT,增强细胞迁移及侵袭能力,其分子机制可能与提高Twist表达有关。Objective：To investigate whether the overexpression of Oct4B1 gene induces epithelial mesenchymal transition in human colorectal cancer SW480 cells and its possible mechanism.Methods：Experimental group（SW480-Oct4B1） ：Transfection of SW480 cell lines in colorectal cancer with Oct4B1 overexpression plasmid;Control group（SW480-Oct4B1） ：negative control plasmid with G418 resistance.Stably transfected cell lines were obtained by G418 culture medium.The two groups were compared with：（1）Detection of Oct4B1 gene expression in stably transfected cell lines by RT-q PCR;（2）Scratches and Transwell assays were used to estimate migration and invasion;（3）Detection of EMT related markers E-cadherin,N-cadherin and Vimentin protein expression by Western blot assay;（4）Detection of Twist gene and protein expression by RT-PCR and Western blot assays.Results：The transient transfection was confirmed by RT-q PCR and the stable transfected cell lines were obtained from two groups of cells transfected with G418 culture medium.Compared with the control group：（1）RT-q PCR revealed increased expression of Oct4B1 gene in the experimental group（P〈0.01）;（2）Cell migration and invasion were significantly increased（P〈0.01）;（3）Epithelial marker：the expression of E-cadherin protein was significantly decreased（P〈0.01）,interstitial marker：the expression of N-cadherin and Vimentin protein was significantly increased（P〈0.01）;（4）Twist mRNA and protein expression were significantly increased（P〈0.01）.Conclusion：Overexpression of Oct4B1 gene can induce epithelial mesenchymal transition in human colorectal cancer SW480 cells,its molecular mechanism may be related to the promotion of Twist expression.

关 键 词：结直肠癌细胞 Oct4B1 上皮间质转化 TWIST 

分 类 号：R735.35[医药卫生—肿瘤]