过表达FLJ22349促进人胚胎肾细胞HEK293及乳腺癌细胞MCF-7的生长  

Effect of hypothetical protein FLJ22349 overexpression on growth of HEK293 cell and breast cancer cell lines

作　　者：戴亚云[1] 赵玫[1] 余权[1] 方明镜[1] 时岚[1] 罗清[1] 袁兴华[1] 黄常志[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤医院肿瘤研究所病因及癌变研究室,北京100021

出　　处：《现代肿瘤医学》2008年第10期1659-1663,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(No.30570904)

摘　　要：目的:人假想蛋白FLJ22349基因位于染色体22q13.2上,编码一个229个氨基酸的蛋白。表达谱分析发现多种肿瘤组织中有FLJ22349的表达,预测该基因可能与肿瘤的发生发展有密切关系。本研究旨在用生物信息学方法分析FLJ22349的结构和功能,并初步研究FLJ22349在HEK293及乳腺癌细胞中的作用。方法:运用各种生物信息学数据库初步分析FLJ22349的结构和功能。逆转录聚合酶链反应(RT-PCR)方法分析FLJ22349基因在小鼠正常组织,人癌症细胞株及HEK293中的表达情况。将载有FLJ22349的基因的质粒转染入HEK293,MCF-7,MDA-MB231中,观察该基因的亚细胞定位。MTT检测细胞的生长变化。结果:FLJ22349基因在人类、猩猩、恒河猴、褐家鼠、小鼠和狗中有很高的同源性,但与其它已知的基因无同源性。小鼠正常组织中有FLJ22349表达;乳腺癌细胞株MDA-MB231,肝癌细胞株Bel-7402也有表达;主要定位于细胞核;转染FLJ22349的HEK293,MCF-7细胞株生长曲线显示促进细胞生长(P<0.05)。FLJ22349转染120h后,MTT实验观察到HEK293,MCF-7细胞的增殖率分别为299%,456%;而对照组分别为233%,272%。结论:FLJ22349基因过表达促进HEK293和MCF-7细胞的生长。Objective：Human hypothetical protein FLJ22349 is located on chromosome 22q13.2 and encodes 229 amino acids. Biological information analysis suggested wide expression profile of FLJ22349 in human cancer tissues. This study was to predict the biological information of human FLJ22349 by Bioinformatics software and investigate the basic effect on cell lines. Methods： Prediction the function of FLJ22349 was examined through different biomedical databases. The expressions of the FIJ22349 mRNA in mouse tissues, human cancer cell lines and HEK293 were detected by reverse transcription polymerase chain reaction（ RT -PCR） ; The recombinant eukaryotic expression vector containing widetype FIj22349 was transfected into HEK293, MCF-7, MDA- MB231 cell lines by cation polymex. The stable transfected cells were selected by G418 and amplified. The effect of F1,122349 on cell growth was assessed by MTT assay. Subcellular distribution of human FLJ22349 was detected under fluorescent microscope. Resuits： F1,122349 was conserved across diverse species, including human, chimpanzee, rhesus, rat, mouse, dog, and cow, and shared no obvious homology to any known genes or protein. RT - PCR analysis revealed FLJ22349 gene expression in many mouse tissues and human cancer cell lines. FLJ22349 was mainly localized in the nucleus. The cell growth rates in HEK293/ pcDNA3.1 （ ＋ ） - FLJ22349 and MCF - 7/ pcDNA3.1 （ ＋ ） - FLJ22349 cell lines were higher than HEK293/pcDNA3.1 （ ＋ ） and MCF - 7/ pcDNA3, 1 （ ＋ ） respectively （ P 〈 0.05 ）. After 120h FIj22349 transfection, the growth rates of HEK293 and MCF -7 were 299% ,456% while those control group were 233% ,272%. Conclution： FLJ22349 is a protein that increases cells growth whn over expressed in HEK293 and MCF- 7 cell lines.

关 键 词：乳腺肿瘤细胞系 HEK293 FLJ22349基因 细胞生长 

分 类 号：R737.9[医药卫生—肿瘤]