摘要
目的:克隆白细胞介素-7(IL-7)剪接变异体,筛选出在IL-7开放阅读框内剪接变异体,为其生物活性的研究打基础。方法:运用自行设计的IL-7引物,采用逆转录聚合酶链反应(RT-PCR)技术,分离正常培养的肝癌细胞株HepG2、结肠癌细胞株HT29IL-7mRNA,再对电泳所获条带进行克隆、测序和分析。结果:肝癌细胞株HepG2、结肠癌细胞株HT29都能检测到IL-7mRNA,而且从培养癌细胞株分离出多条新带,经亚克隆及测序,证实这些条带是来源人IL-7基因、但缺乏不同外显子的IL-7剪接变异体cDNA;通过和IL-7cDNA比对分析,发现其中5个剪接变异体在IL-7开放阅读框内。结论:IL-7剪接变异体广泛存在于各种肿瘤细胞中。对其进一步的研究,将为癌症病人的免疫调节紊乱和临床免疫治疗提供新的研究方向。
AIM： To clone and select the splice variants in open reading frame of interleukin 7 from cancer cell lines.METHODS： Reverse transcription-polymerase chain reaction（RT-PCR） was used for investigating the expression of IL-7.According to IL-7 gene cDNA,the primers were designed and synthesized,then the splice variants of IL-7 in human cancer cell lines was identified,cloned into vector and sequenced.RESULTS：IL-7 mRNA can be detected in Hepatic cancer cells and Colon cancer cells.A number of new bands of IL-7 were obtained from cultured cancer cell lines,which lack of different exons.By analyzing the splice variants,we found 5 of them in open reading frame of IL-7.CONCLUSION： Splice variants of IL-7 are expressed in many tumor cell lines.Further investigation of the Splice variants can provide a new research direction for cancer patient immune disorders and clinical immunotherapy.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2012年第4期367-370,共4页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
中国-意大利科技合作项目(200625)
关键词
白细胞介素7
克隆
序列分析
肿瘤细胞
剪接体
Interleukin 7
Cloning
Sequence analysis
Tumor cells
Splice variants
