HGPRT缺陷型Lewis肺癌细胞株的筛选及其生物学特征

HPRT-defective Lewis lung carcinoma cell line and its biological characteristics

目的 为细胞融合等实验提供次黄嘌呤鸟嘌呤磷酸核糖转移酶 (HGPRT)缺陷细胞株。方法 用 8 氮杂鸟嘌呤 ( 8 AG)对Lewis肺癌细胞株L3 8进行长期渐进式给药 ,从中克隆筛选出HGPRT缺陷细胞株 ,并观察其体内成瘤和肺转移特性。结果 经过 1年的筛选鉴定 ,获得了在 2 0mg/L 8 AG和 16 40培养液中能长期稳定生长 ,在HAT选择性培养基中不能形成克隆的AL990 1细胞株。AL990 1和L3 8的染色体众数分别为5 8和 6 2 ,在C5 7BL/ 6小鼠体内成瘤率均为 10 0 % ( 10 / 10 ) ,肺转移率分别为 30 % ( 3/ 10 )和 70 % ( 7/ 10 )。结论 HGPRT缺陷型AL990 1细胞株基本保持了L3 8细胞的生物学特征 ,可作为制备Lewis肺癌DC融合疫苗的亲本抗原细胞。

Objective To provide an HGPRT (hypoxanthine guanidine phosphoribosyl transferase) defective cell line for the establishment of lung cancer and dendritic cell(DC) fused vaccine. Methods The HGPRT defective cell line was gradually induced by 8 AG (8 azaguanine) from the Lewis lung carcinoma cell line(L3 8). Its biological characteristics and tumorigenicity were observed in vivo and in vitro. Results The HGPRT defective cell line, AL9901, was obtained after one year of selective culture and identification. AL9901 cell line could grow steadily in 20?mg/L 8 AG, but not in HAT selective medium. The chromosome modal numbers of AL9901 and L3 8 cell line were 58 and 62 respectively, lung metastatic rates were 30%(3/10) and 70%(7/10) respectively, and their tumorigenic rates were both 100%(10/10). Conclusion The HGPRT defective cell line, AL9901, maintains the biological characteristics and carcinogenicity of the L3 8 cell line. It can be used as a parental antigenic cell for the establishment of Lewis lung carcinoma DC fused tumor vaccine.