HGPRT缺陷性人结肠癌细胞株的建立

THE ESTABLISHMENT OF HGPRT MUTANT FROM HUMAN COLORECTAL CARCINOMA CELL LINE(CCL187)

本研究首先用化学突变剂乙基甲磺酸(EMS)处理人结肠癌细胞系CCL187,然后用嘌呤类似物8-氮鸟嘌呤(8-AG)筛选次黄嘌呤鸟嘌呤转磷酸核糖酶缺陷细胞(HGPRT^-),历时半年,成功获得抗8-AG毒性突变体,即HG-PRT缺陷细胞株MCL187。该缺陷株细胞HG-PRT酶活性显著低于野生型细胞,在8-AG中能够存活,在HAT选择性培养基中则死亡,与野生型细胞同样表达结肠癌相关抗原LEA。此缺陷株的成功建立,为杂交细胞制备及其应用研究奠定了基础。

The wild type of human colorectal carcinoma CCL187 cells were treated firstly by a chemical carcinogen EMS for the induction of mutagenesis.then the mutant cells were selected in the medium containing 8-AG at the concentration of 20μg/ml. After six months' screening.the HGPRT mutant line MCL187 was established. It was found that the mutant celis could grow vigorously in the medium containing 8-AG(20μg/μl)but not in HAT medium. The HGPRT assay showed a significant quantitative difference between the HGPRT mutant cells and the witd type. Enpression of tumor associated antigen LEA on the HGPRT mutant celis was similar to that on the wild type.