自噬途径对反式-BPDE诱导人支气管上皮细胞16HBE影响及其机制的探讨

Role of autophagy in the malignant transformation of 16HBE cells by anti-BPDE

目的:研究反式-BPDE诱导正常的人支气管上皮细胞16HBE发生恶性转化前后自噬水平的变化及其可能机制。方法:利用前期建立的细胞恶性转化模型,采用激光共聚焦扫描显微镜成像技术,在无损伤状态下,实时观测反式-BPDE恶性转化细胞(16HBE-T)和正常细胞(16HBE-N)中自噬体标记蛋白GFP-LC3的荧光强度及定位,并利用蛋白质印迹法技术检测细胞内自噬信号通路关键效应分子的表达。结果:16HBE-T细胞与16HBE-N细胞相比,GFP-LC3蛋白的点状聚集明显减少,只有16HBE-N细胞的1/3左右,表明自噬水平下降,蛋白质印迹法检测发现,mTOR激酶活性上升,Beclin 1表达下降。结论:反式-BPDE可能通过自噬途径参与诱导16HBE细胞恶性转化的癌变过程,将为预防和治疗肺癌提供重要信息。

OBJECTIVE：To investigate the mechanism of autophagy-related signaling pathway in the anti-BPDE trans- formed 16 HBE （16 Ht^E-T） cells, and find new access for prevention and treatment of lung cancer. METHODS： By using of the pre-established model of malignant transformation of cells, we monitored the position and fluorescence intensity of the GFP-LC3 protein that was used to show autophagy in living cells with confoeal laser scanning microscopy imaging. We also detect the chan- ges of autophagy in the normal 16HBE （16HEE-N） and malignant transformed cells （16HBE-T） with Western blotting. RE- SULTS：Compared with that of 16HBE-N ceils, the GFP-LC3 protein punctuate aggregation of 16HBE-N cells significantly re- duced to 1/3 amount of 16HBE-N cells,suggesting that autocracy levels declinded. At the same time,the roTOR kinase activity and Beclin 1 expression were found to be increased and decreased, respectively. CONCLUSION： This experiment demonstrates that autophagy involves in the malignant transformation induced by anti-BPDE via multiple pathways, and the study of autophagy will be great benefit for the prevention and treatment of lung cancer.