CAJ | 학술논문

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Western blot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.
목적 탐토JWA대완화제N-갑기-N'-초기-N-아초기고(MNNG)유도인지기관상피세포(HBE)악성전화적영향급가능궤제.방법 건립JWA고표체HBE세포주,용MNNG유도HBE세포악성전화,용새서람(MTT)법검측MNNG유도후세포생장상황,용연경지집락시험검측세포비묘착의뢰생장능력,용Western blot법검측P53단백적표체변화규률.결과 경MNNG처리적정상HBE세포악성전화후생장속도명현쾌우고표체JWA적HBE세포화미경MNNG처리적HBE세포,차이유통계학의의(P＜0.05).경MNNG처리적JWA고표체적HBE세포화미용MNNG처리적HBE세포적극륭형성솔(8.06%화10.14%)명현저우MNNG처리적악성전화적HBE세포(26.80%),차이유통계학의의(P＜0.01).MNNG유도정상적HBE세포악성전화과정중,P53단백축점증가;이재JWA고표체HBE세포,경MNNG처리후,P53단백조기(제1～2대)유일과성적표체증고,차후,수착전대수증가,P53표체칙축점하강,세포최종미현시악성전화표형특정.결론 JWA가능통과P53단백표체조절MNNG유도HBE세포적악성전화.
Objective To investigate the role and possible mechanism of JWA in N-methyl-N'-nitroN-nitrosoguanidine (MNNG) inducing human bronchial epithelial (HBE) cells' neoplastic transformationMethods JWA overexpression vector and its stable trasfection HBE cells were established. The characteristics of transformed HBE cells were determined by methyl thiazolyl tetrazolium (MTT) assay and the soft agar colony formation 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 of JWA overexpression HBE cells with and without MNNG treatment(8.06% and 10.14% ) was significandy lower than that of the norreal HBE cells with MNNG treatment (26.80%)(P＜0.01). After exposure of MNNG,the P53 expressions weregradually 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 spoctrum with increased passages of the cells.Conclusion JWA might be a key molecule and play an important role in MNNG inducing neoplastic transformarion in HBE cells through regulation of the expression of P53.