RNA干扰技术抑制锌指蛋白139对胃癌原位移植裸鼠血清的影响  

Proteomics research about effect of zinc finger protein 139 knockdown with RNA interference on se- rum of orthotopic transplantation nude mouse with gastric cancer

作　　者：范立侨[1] 李勇[1] 赵群[1] 檀碧波[1] 陈茂良[1] 王冬[1] 赵雪峰[1] 郝英杰[1] 

机构地区：[1]河北医科大学第四医院外三科,石家庄050011

出　　处：《中华实验外科杂志》2013年第7期1456-1458,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81072033）;河北省自然科学基金资助项目（C2010000619）;河北省普通高校强势特色学科资助项目[冀教高（2005）52];河北省卫生厅科研基金资助项目（20110460）

摘　　要：目的观察抑制人胃癌细胞株SGC7901中锌指蛋白139（ZNFl39）表达对胃癌原位移植裸鼠血清蛋白质的影响，探讨ZNFl39参与胃癌进展的机制。方法构建针对ZNF139的小干扰RNA（siRNA）重组质粒并导入胃癌细胞SGC7901；G418筛选出稳定转染细胞克隆并经皮下瘤鼠间传代6代后应用OB生物胶粘贴法进行原位移植；应用荧光双向差异凝胶电泳（2D—DIGE）及液质联用（LC—MS）技术鉴定差异蛋白点，并应用Westernblot法验证鉴定出的部分蛋白质。结果胃癌原位移植裸鼠成瘤率100％（15／15）。siRNA重组质粒抑制了ZNFl39在SGC7901中的表达，siRNA—ZNFl39组胃癌原位移植裸鼠瘤体生长明显减慢。在转染组和对照组原位移植裸鼠血清中鉴定出5种差异蛋白质，抑制ZNFl39后表达下调的为载脂蛋白E（APOE）、激肽原1（KNGl），表达上调的有α2-巨球蛋白（α2-MG）、线粒体转录终止因子（mTERF）、DIXDCl。Westernblot法对KNG1、mTERF、APOE的表达进行了验证，结果与蛋白质组学一致。结论ZNF139可能通过促进血清APOE、KNGl及抑制mTERF的表达来参与胃癌进展。Objective To observe the effect of inhibiting zinc finger protein 139 （ZNF139） on se- rum proteins in orthotopic transplantation nude mice model with gastric cancer, and to investigate the mech- anism for ZNF139 involved in progression of gastric cancer. Methods Small interfering RNA （siRNA） re- combinant plasmid targeting ZNF139 gene was constructed and imported into gastric cancer cell line SGC7901. After screened stably transfected cells by G418, gastric cancer cells were injected into nude mice subcutaneously. Transplanted tumor tissue was constructed through repeatedly subcutaneously inocula- ting and passaging between nude mice, and 6th-generation subcutaneous tumor was pasted with OB biologi- cal glue orthotopically on the stomach wall of BALB/c nude mice. The proteins in nude mice serum sam- ples were separated by fluorescence two-dimensional difference gel electrophoresis （2D-DIGE） and the dif- ferential proteins were identified by liquid chromatography-mass spectrometry （LC-MS）. Parts of the pro- teins were identified by Western blotting. Results The tumor-forming rate in orthopotic implantation was 100% （15/15）. ZNF139 was inhibited obviously, and the tumor growth of orthotopic transplantation nude mice model was obviously slower in siRNA-ZNF139 group. Five differential proteins were identified in or- thotopic transplantation nude mice serum samples. Results showed that apolipoprotein E （APOE） and the kininogen 1 （ KNG1 ） were down-regulated in gastric cancer orthotopic mouse serum, and mTERF was up- regulated after siRNA-ZNF139 interference. Results of Western blotting were consistent with proteomics. Conclusion ZNF139 in gastric cancer may up-regulate serum levels of APOE and KNG1, while down-reg- ulate mTERF to participate in the progression of gastric cancer.

关 键 词：胃癌原位移植 锌指蛋白139 蛋白质组学 

分 类 号：R735.2[医药卫生—肿瘤]