人IL-15基因在CHO细胞中的表达及其对鼻咽癌细胞增殖的抑制作用  

Expression of Human IL-15 Gene in CHO Cells and its Inhibitory Effect on Proliferation of Nasopharyngeal Carcinoma Cell Line in Vitro

作　　者：何跃平[1] 张艳[2] 郑家法[1] 申海荣[1] 田梦秋[1] 梁贵玲[1] 

机构地区：[1]南华大学附属南华医院耳鼻咽喉科,湖南衡阳421002 [2]南华大学医学微生物学与免疫学教研室

出　　处：《实用预防医学》2007年第3期653-655,共3页Practical Preventive Medicine

摘　　要：目的在CHO细胞中进行人白细胞介素-15基因真核表达载体的稳定表达,并检测重组表达蛋白rhIL-15对鼻咽癌细胞体外增殖的抑制作用。方法将真核表达载体pcDNA3.1(+)-hIL-15转染CHO细胞,G418筛选稳定表达株;采用RT-PCR和ELISA法对hIL-15的表达进行检测,MTT法检测rhIL-15对人鼻咽癌细胞系CNE-2Z体外增殖的抑制效应。结果人IL-15在CHO细胞中获得稳定表达,RT-PCR显示细胞的rhIL-15mRNA呈现高水平,ELISA检测到rhIL-15在细胞中的分泌性表达,浓度达(3.08±0.23)μg/ml,且所表达的人IL-15具有较强的抑制CNE-2Z鼻咽癌细胞增殖的作用。结论成功地在CHO细胞中获得了hIL-15蛋白的高效、稳定表达,重组表达蛋白rhIL-15体外能有效地抑制鼻咽癌细胞增殖,为进一步研究人IL-15抗肿瘤的分子机制及潜在的应用奠定了基础。Objective The eukaryotic expression vector of hlL- 15 cDNA was expressed in Chinese hamster ovary （CHO） cells, and the inhibition effects of recombinant hlL- 15 protein on Nasopharyngeal carcinoma cell line proliferation in vitro were detected. Methods The recombinant expression plasmid pcDNA3.1 （ ＋ ） - hlL - 15 was transfected into CHO cells and the positive clone was screened by G418. Then the expression of hlL - 15 in CHO cells was detected by RT- PCR and ELISA. The inhibitory effects of recombinant protein hlL- 15 on proliferation of human nasopharyngeal carcinoma cell line ONE- 2Z were tested by MTT assay. Results Human IL - 15 cDNA was successfully expressed in CHO cells. Stable expression of hlL- 15 in CHO cells was identified with RT- PCR, the secretion expression of hlL- 15 was proved by ELISA and its concentration was 3.08 ± 0.23 μg/ml. The hlL - 15 showed a strong suppressive effect on proliferation of CNE - 2Z cells. Conclusion CHO system which expresses stable and high- level hlL- 15 is established successfully, the recombinant protein hlL- 15 has an effective inhibitory effect on proliferation of CNE - 2Z cells in vitro, which lays the foundation for the further study of molecular mechanism of hlL- 15 on anti - tumor activity and other ootential aoolication.

关 键 词：人自细胞介素-15 稳定转染 CHO细胞 鼻咽癌细胞 生长抑制 

分 类 号：R739.6[医药卫生—肿瘤]