JWA对N-甲基-N’-硝基-N-亚硝基胍诱导人支气管上皮细胞恶性转化的影响

JWA regulates N-methyl-N＇-ultro-N-ultrosoguauldine induced malignant transformation in human bronchial epithelial cells

目的 探讨JWA对烷化剂N-甲基-N’-硝基-N-亚硝基胍（MNNG）诱导人支气管上皮细胞（HBE）恶性转化的影响及可能机制。方法建立JWA高表达HBE细胞株，用MNNG诱导HBE细胞恶性转化，用噻唑蓝（MTT）法检测MNNG诱导后细胞生长状况，用软琼脂集落试验检测细胞非锚着依赖生长能力，用Westernblot法检测P53蛋白的表达变化规律。结果经MNNG处理的正常HBE细胞恶性转化后生长速度明显快于高表达JWA的HBE细胞和未经MNNG处理的HBE细胞，差异有统计学意义（P〈0．05）。经MNNG处理的JWA高表达的HBE细胞和未用MNNG处理的HBE细胞的克隆形成率（8．06％和10．14％）明显低于MNNG处理的恶性转化的HBE细胞（26．80％），差异有统计学意义（P〈0．01）。MNNG诱导正常的HBE细胞恶性转化过程中，P53蛋白逐渐增加；而在JWA高表达HBE细胞，经MNNG处理后，P53蛋白早期（第1～2代）有一过性的表达增高，此后，随着传代数增加，P53表达则逐渐下降，细胞最终未显示恶性转化表型特征。结论JWA可能通过P53蛋白表达调节MNNG诱导HBE细胞的恶性转化。

Objective To investigate the role and possible mechanism of JWA in N-methyl-N＇-nitro- N-nitrosoguanidine （ MNNG ） inducing human bronchial epithelial （ HBE ） cells＇ neoplastic transformation. Methods JWA overexpression vector and its stable trasfection HBE cells were established. The characteris- tics of transfornled HBE cells were determined by methyl thiazolyl tetrazolium （MTT） assay and the soft agar colony fornlation assay. The expressions of JWA and P53 were detected by Western blot. Results The growth rates of the HBE cells which were treated with MNNG were significantly accelerated than the JWA overexpression HBE cells and controlled HBE cells （P〈0.05）. The soft agar colony formation ofJWA overexpression HBE cells with and without MNNG treatment（8.06% and 10.14% ） was significantly lower than that of the normal HBE cells with MNNG treatment （ 26.80% ） （P〈0.01）. After exposure of MNNG, the P53 expressions were gradually increased in HBE cells with the increased passages. However,the expression of P53 in JWA over expressed HBE cells showed a different manner. P53 reached an over expression peak at early stage（the first passage）,and then with a gradually down-regulated expression spectrum with increased passages of the cells. Conclusion JWA might be a key molecule and play an important role in MNNG inducing neoplastic transformation in HBE cells through regulation of the expression of P53.