摘要
目的探讨人支气管上皮细胞（BEAS-2B细胞）恶性转化过程中生物学特性变化及p53动态表达的意义。方法以500μg/mL的烟草特有亚硝胺4-（N-亚硝基甲氨基）-1-（3-吡定基）-1-丁酮（NNK）处理BEAS-2B细胞24 h后,细胞在体外连续传代培养,在此过程中观测细胞生物学特性,以免疫组化法检测p53表达的动态变化。结果第5代细胞显示抗血清生长,细胞在裸鼠体内不成瘤;第15代细胞在半固体琼脂中克隆形成率（0.32‰）为对照细胞（0.023‰）的13.9倍,细胞在裸鼠体内不成瘤。细胞超微结构显示转化细胞特征;第25代细胞在裸鼠体内成瘤,组织学证实为鳞癌（Ⅰ~Ⅱ级）。细胞超微结构显示具有明显的肿瘤细胞特征,对照组各代BEAS-2B细胞p53表达均为阴性。各代BEAS-2BNNK细胞均有不同程度的表达,随着传代次数增加表达呈明显上升趋势,各代间差异有统计学意义（P〈0.001）。结论浓度为500μg/mL的NNK可成功诱发BEAS-2B细胞恶性转化,BEAS-2B细胞生物学特性改变是一个多步骤、多阶段的发展过程,p53蛋白在BEAS-2B细胞转化成癌细胞以前已有较明显表达,可能有助于吸烟高危人群罹患肺癌的预警及普查。
Obiectlve To study the change of biological characteristics and significance of p53 expression of BEAS-2B cells during malignant transformation.Methods BEAS-2B cells treated with NNK at the oonceatration of 500μg/mL for 24 h were subcultured continuously in vitro and their biological characteristics and ultrastructure were observed.The change p53 expression of BEAS-2B cells were detected by immunohistochemical method.Results The serum resistance was significantly increased in the 5th passage BEAS-2BNHK cells and could not grow into tumor in nude mice.The plating efficieney of the 15th passage BEAS-2BNNK ceIls in soft agar（0.32‰）increased 13.9 fold compared with that of control cells（0.023‰）.The cells had the biological characteristic of transformation cells but could not grow into tumor in nude mice.The 25th passage could grow into tumor in nude mice.The tumor was squamous cell carcinoma in morphology confirmed by histopethologlcal examination.The ultrastructrure showed that BEAS-2BNNK cells were transformed into cancer cells.p53 protein expression of BEAS-2B cells was negative,but positive in the different passage BEAS-2BNNKcells.With the passage of BEAS-2BNNK cells increasing gradually,p53 protein expression rised more and more obviously（P〈0.001）.Condusion The model of malignant transformation of BEAS-2B cells induced by NNK（500 μg/mL）can be created successfully.During the period,the change of bicological characteristic and ultrastructum indicated that the malignant transformation of BEAS-2BNNK cell is a multiple-step course.The characteristic of p53 being expressed before BEAS-2BNNK cells changed into cancer cells is beneficial to early-warning and mass screening of lung cancer in high-risk smoking population.
出处
《重庆医学》
CAS
CSCD
北大核心
2010年第17期2275-2277,F0002,共4页
Chongqing medicine
作者简介
广东中山医科大学在读博士生。
