两种体外培养条件下肺腺癌A549细胞生物学特性的变化

目的 比较体外肺腺癌A549细胞在平面及立体培养条件下细胞周期、细胞凋亡及对化疗药物敏感性的变化。方法 采用旋转培养瓶结合台式水浴恒温振荡器旋转培养的方法，进行肺腺癌A549细胞体外立体培养，并通过流式细胞技术、原位细胞凋亡检测试剂盒、四唑盐(MTT)比色法及血球计数器计数法比较两种培养条件下肺腺癌A549细胞在细胞周期、细胞凋亡及对阿霉素(ADM)敏感性方面的差异。结果 通过倒置显微镜、电镜观察及照相，确定了体外三维立体培养模型的建立；同平面培养比较，立体培养细胞存在细胞周期中G1期阻滞现象、细胞凋亡率低，对阿霉素的敏感性低的特征。结论 腺癌A549细胞在两种体外培养条件下生物学特性存在明显差异，三维立体细胞培养模型更接近实体瘤的体内实际情况。

The changes in biological characteristics of lung adenocarcinoma A549 cells under two in vitro culture systems

Objective To investigate the changes in cells on cell cycle,cell apoptosis and susceptibility to chemotherapeutic drugs of lung adenocarcinoma A549 cells under two dimensional (2D system) and three dimensional (3D system) culture conditions. Methods The three dimensional culture of A549 cells was carried out using rotatable culture bottle and desk-top water bath shake. Flow cytometer,in situ cell apoptosis detection kit,MTT and blood cell counter were employed to compare cell cycle,cell apoptosis and susceptibility to ADM of A549 cells under different culture conditions. Results The three dimensional culture system was confirmed to have been successful by the aid of observation under inverted and electron microscope. There were significant retardation of G_ 1 phase,lower cell apoptosis rate and decreased susceptibility to ADM in A549 cells in 3D system compared with those in 2D system. Conclusions There were remarkable differences in biological characteristics of A549 cells in two culture systems,indicating that A549 cells cultivated in 3D system simulated better solid tumor in vivo . The 3D system was very useful for further investigation of the behavior of solid tumor,so that anti-carcinoma chemotherapeutic drugs could be advantageously tested in vitro before clinical application.