人Sep15的基因克隆和功能的初步研究  

Redox Reactions of Sep15 and Its Relationship with Tumor Development

作　　者：吴红杰[1] 林晨[1] 查园园[1] 杨建国[1] 张明诚[2] 张雪艳[1] 梁肖[1] 付明[1] 吴旻[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [2]中国预防医学科学院病毒研究所肝炎病毒研究室,北京100052

出　　处：《癌症》2003年第2期119-122,共4页Chinese Journal of Cancer

摘　　要：背景与目的:Sep15是1998年发现的一种新的硒蛋白。文献报道Sep15可能与肿瘤的发生有关,并参与二硫键的还原。但迄今为止,Sep15的确切功能还不清楚。本实验拟对Sep15与肿瘤的关系及其抗氧化功能进行初步研究。方法:应用RT-PCR方法获得人Sep15的全长cDNA基因,并重组到真核表达载体pcDNA3.1(+)上,将重组质粒pcDNA3.1-Sep15转染到人肝癌细胞株BEL-7402,得到Sep15高表达的细胞株BEL-7402-Sep15,然后通过一系列体内和体外实验研究Sep15与肝癌细胞的关系及其抗氧化作用。结果:转染Sep15基因不影响肝癌细胞株BEL-7402的细胞形态、生长速率、克隆形成能力和裸鼠体内肿瘤生长速率;检测不同浓度外源性H2O2氧应激处理后3种细胞BEL-7402-Sep15、BEL-7402-pcDNA3.1和BEL-7402的细胞存活率,结果显示BEL-7402-Sep15的细胞存活率明显高于BEL-7402-pcDNA3.1和BEL-7402(P<0.05)。结论:转染Sep15基因的肝癌细胞株BEL-7402具有抗氧化的生物学活性。BACKGROUND & OBJECTIVE: Sep15 is a selenium containing protein identified in 1998. This protein may be involved in cancer etiology and it may have redox function. The objective of this study was to investigate the relationship between the redox function of Sep15 and tumor development. METHODS:The full length DNA sequence of Sep15 was obtained by RT PCR and then recombined to eukaryotic expression vector pcDNA3.1(+). The BEL 7402 Sep15 cell line, which expressed the high levels of Sep15 by transfecting the cultured hepatocarcinoma cell line BEL 7402 with pcDNA3.1 Sep15 was generated. From morphologic investigation, cell growth curve, clone formation and nude mice tumor growth curve, the relationship between Sep15 and hepatocarcinoma cell line BEL 7402 was determined. Furthermore, the redox reaction of sep15 was detected by MTT assay. RESULTS:There was no distinct effect of transfection of Sep15 gene on BEL 7402 Sep15 cell. The cell survival rate was drastically different between BEL 7402 Sep15 cell and both BEL 7402 pcDNA cell and BEL 7402 Sep15 cell after foreign H2O2 reactive oxygen stress (P< 0 05). CONCLUSION:Transfecting Sep15 gene did not influence the growth characteristics of BEL 7402 cell line and Sep15 may have redox function.

关 键 词：Sep15 硒蛋白 抗氧化 肿瘤 克隆 

分 类 号：R730.2[医药卫生—肿瘤]