不同培养方式中肺腺癌细胞系A549表达E-钙粘素的差异及其意义

目的初步探讨E-钙粘素在肿瘤群集耐药中的可能机制.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,进行肺腺癌细胞A549体外立体培养,同时进行普通平面培养,通过流式细胞技术、Western blot、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检测细胞周期,E-钙粘素的表达,细胞凋亡率及对阿霉素(ADM)敏感性.结果①立体培养细胞存在E-钙粘素的高表达;②同平面培养比较,立体培养细胞出现细胞周期G1期阻滞、细胞凋亡率低、对ADM敏感性低等表现,给予E-钙粘素阻滞剂SHE78-7后,两者差异明显缩小.结论 E-钙粘素介导的细胞-细胞粘附作用可通过调控细胞周期以及细胞凋亡,影响肺腺癌A549细胞对ADM的敏感性,在肿瘤群集耐药中发挥作用.

Differences in E-cadherin expression of lung adenocarcinoma cell line A549 under different culture conditions and the related significance

Objective To investigate the roles of E-cadherin on multicellular resistance of lung adenocarcinoma cell line A549. Methods Three dimensional culture of A549 cells was achieved using rotatable culture bottle and desk-top water bath shake. Flow cytometry, Western blot, in situ cell apoptosis detecting kit, MTT, and blood cell counter were employed to detect the expression of E-cadherin, cell apoptosis, and susceptibility to ADM of A549 cells under different culture conditions. Results Compared with cells of two dimensional culture system (2D system), higher E-cadherin expression was observed in cells of three dimensional culture system (3D system), and G 1 phase arrest, lower cell apoptosis rate, and decreased susceptibility to ADM were found in cells of 3D system. The differences diminished significantly after using SHE78-7 as the blocker of E-cadherin. Conclusion E-cadherin-mediated enhancement of intercellular adherence can function in multicellular resistance of lung adenocarcinoma A549 cells through influencing cell cycle, apoptosis, and susceptibility to ADM.