人肝癌细胞系HepG2的次黄嘌呤鸟嘌呤磷酸核糖转移酶缺陷型的建立  

Establishment of Hypoxanthine Guanine Phosphoribosyl Transferase Mutant from Human Liver Cancer Cell Line HepG2

作　　者：特日格乐[1] 邰大鹏[1] 刘刚[1] 道日娜[1] 王蓉蓉[1] 郭惠东[1] 李瑶[1] 李煜[1] 

机构地区：[1]内蒙古大学哺乳动物生殖生物学与生物技术教育部重点实验室,内蒙古呼和浩特010021

出　　处：《中国畜牧兽医》2011年第12期92-95,共4页China Animal Husbandry & Veterinary Medicine

基　　金：内蒙古自治区自然科学基金项目(2009MS0503);国家大学生创新性实验计划项目(101012618)

摘　　要：本试验旨在建立次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)缺陷型的HepG2细胞系,为细胞杂交与单克隆抗体的研究提供亲本细胞。通过N-甲基-N-硝基-N-亚硝基胍(MNNG)诱导HepG2细胞突变,逐步提高培养液中6-巯基鸟嘌呤(6-TG)的浓度,筛选出对6-TG有抗性的细胞株,检测它在HAT中生长的情况。得到可在含20μg/mL 6-TG的高糖DMEM完全培养基中稳定生长的细胞株,此细胞株在HAT培养基中第3天出现明显的死亡,第12天全部死亡,诱变后细胞的染色体分布于32-88,染色体众数为48。突变筛选出的细胞具有对6-TG抗性和对HAT敏感的特性,证实该细胞是HGPRT缺陷型细胞株,为以后进行杂交瘤制备提供了适宜的亲本细胞。To establish a hypoxanthine guanine phosphoribosyl transferase（HGPRT） deficient cell line from HepG2 cell line,so as to provide parent cells for hybridoma and monoclonal antibody study.The HepG2 cells were cultured with N-methyl-N-nitro-N-nitrosoguanidine（MNNG） for mutation and selected by increasing concent ration of 6-thioguanine（6-TG）.The 6-TG resistant mutant cells were cultured in the medium containing 6-TG or HAT.After four months of screening,a deficient cell line was established.It could grow vigorously in the medium containing 20 μg/mL 6-TG,while it all died within 12 days in the medium containing HAT.Karyotypic analysis showed that the number of the chromosomes of the deficient cells is between 32 and 88,with the modal number being 48.The 6-TG resistant and aminopterin-sensitive character of the mutant cell line reveals that it is deficient in HGPRT.The cell line established in this study will be a good parent cell line for preparation of hybridoma in the future.

关 键 词：细胞诱变 HEPG2 HGPRT缺陷型 

分 类 号：S852.3[农业科学—基础兽医学]