人支气管上皮细胞c-Ha-ras^(V12G)基因转染细胞系促癌剂检测模型的建立

Development of tumor promoter testing model with transfected human bronchial epithelial cells harboring c-Ha-ras^(V12G)

目的 :建立一个新的促癌剂检测模型。 方法 :采用电穿孔法 ,将人类突变的 c- Ha- ras V1 2 G基因转入人支气管上皮细胞BEAS- 2 B中 ,将获得的 G418抗性 (G418r)单克隆细胞株用佛波醇酯 (PMA)处理 ,并选用克隆形成率、锚着独立性生长及血清抗性等表型改变对受试细胞进行了恶性程度检测。 结果 :(1)在促癌剂 PMA作用下 ,BEAS- 2 B基因转染细胞发生了恶性转化 ;(2 )表达突变 ras基因的上皮细胞 ,在大剂量 PMA(>1μmol)的急性刺激 (2 4h)下 ,可诱导细胞死亡 ;(3)随着细胞恶性程度的增加 ,某些促癌剂的促细胞分化作用将减弱 ,并有生长刺激作用。 结论 :该模型可用于促癌剂的检测及促癌作用机制研究。

Objective: To develop a tumor promoter testing system. Methods: Human mutant c-Ha-ras V12G gene was introduced into human bronchial epithelial cells BEAS-2B by electroporation. After exposure to PMA, the phenotype, such as colony forming efficiency,anchor independent growth and serum resistance of selected G418 r (G418-resistant) monocolonies were observed. Results: (1) The neoplastic transformation of transfected BEAS-2B cells were induced by PMA; (2) The death of human epithelial cells expressing a mutant ras oncogene were induced by PMA at high concentration (>1 μmol/L,24 h); (3) With the development of neoplastic transformation, the resistance to terminal differentiation induced by some promoter increased till the promoter became mitogenic. Conclusion: The development of transfected cells provides a powerful and convenient model for detecting potential tumor promoters and research on the mechanisms of promotion tumor.