Zhao Xiao-Jia, Xu Yan-Qiong, Chen Rui
School of Public Health, Nanjing Medical University, Nanjing 210029, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2008 Jul;26(7):395-400.
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.
JWA overexpression vector and its stable transfection 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.
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 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.
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.
探讨JWA在N-甲基-N'-硝基-N-亚硝基胍（MNNG）诱导人支气管上皮（HBE）细胞恶性转化中的作用及可能机制。
构建JWA过表达载体并建立其稳定转染的HBE细胞。通过甲基噻唑基四氮唑（MTT）法和软琼脂集落形成试验检测转化后HBE细胞的特性。采用蛋白质免疫印迹法检测JWA和P53的表达。
MNNG处理的HBE细胞生长速度比JWA过表达的HBE细胞和对照HBE细胞显著加快（P<0.05）。JWA过表达的HBE细胞无论是否经MNNG处理，其软琼脂集落形成率（8.06%和10.14%）均显著低于经MNNG处理的正常HBE细胞（26.80%）（P<0.01）。MNNG作用后，HBE细胞中P53表达随传代次数增加而逐渐升高。然而，JWA过表达的HBE细胞中P53的表达呈现不同趋势。P53在早期（第一代）达到过表达峰值，随后随着细胞传代次数增加表达谱逐渐下调。
JWA可能是关键分子，通过调节P53表达在MNNG诱导HBE细胞恶性转化中起重要作用。
