二氯化钴诱导唾液腺腺样囊性癌ACC-M细胞自噬的实验研究  

Cobalt chloride induced autophagy in salivary adenoid cystic carcinoma

作　　者：尹长伟[1] 吴海威[2] 黄圣运[1] 马猛[2] 陈占伟[1] 张东升[1,2] 

机构地区：[1]山东大学附属山东省立医院口腔颌面外科,山东济南250021 [2]山东大学口腔医学院山东省口腔生物医学重点实验室,山东济南250012

出　　处：《中国口腔颌面外科杂志》2015年第6期491-496,共6页China Journal of Oral and Maxillofacial Surgery

摘　　要：目的:研究二氯化钴(CoCl_2)对唾液腺腺样囊性癌ACC-M细胞增殖和自噬的影响。方法:应用MTT法对ACC-M细胞进行细胞活力测验,以检测CoCl_2对细胞增殖的抑制作用;透射电镜观察自噬体形成;双免疫荧光标记、实时定量PCR(RT-PCR)及Western蛋白免疫印迹检测自噬相关基因HIF-1α/BNIP3通路相关基因、自噬相关蛋白微管相关蛋白1轻链3(microtubule associated protein 1 light chain 3,LC3)及Beclin 1的表达。采用SPSS15.0软件包对数据进行双尾t检验。结果:CoCl_2可降低ACC-M细胞的活性,同时诱导自噬体的形成;透射电镜下,实验组细胞内可见自噬体样的双层膜结构;RT-PCR及Western蛋白免疫印迹检测显示,CoCl_2可使HIF-1α/BNIP3信号通路和自噬相关蛋白的表达显著提高。结论:CoCl_2可模拟低氧环境,从而诱导ACC-M细胞产生自噬,HIF-1α/BNIP3信号通路在这一过程中扮演着重要角色。PURPOSE ： Adenoid cystic carcinoma （ACC） is one of the common types of salivary gland malignancies in the head and neck. Our previous study confirmed that autophagy is closely related to tumorigenesis of ACC. This study investigated the expression of Cobalt chloride （CoCl2）-induced autophagy and its role in tumor invasion. METHODS ： Autophagosome formation and upregulation of autophgy-related microtubule-associated protein 1 light chain 3 （LC3） and Beclin 1 were detected in ACC-M cells in response to CoCl2. HIF-1α/BNIP3 signaling pathway was investigated in hypoxia-induced autophagy in ACC. Statistical comparisons between groups were performed using two-tailed Student’s t test with SPSS 15.0 software package. RESULTS ： CoCl2 reduced cell viability of ACC-M cells, and induced formation of autophagosomes. RT-PCR and Western blot showed that the expression of HIF-1α/BNIP3 signal pathway of autophagy related protein was significantly increased. CONCLUSIONS ： The results revealed that CoCl2 mimetic hypoxia could induce autophagy in ACC, and HIF-1α/BNIP3 pathway plays an important role in activation of hypoxia-induced autophagy in ACC.

关 键 词：二氯化钴 自噬 腺样囊性癌 低氧 HIF-1Α 

分 类 号：R739.87[医药卫生—肿瘤]