Obatoclax induces Beclin 1- and ATG5-dependent apoptosis and autophagy in adenoid cystic carcinoma cells.
Obatoclax induces Beclin 1- and ATG5-dependent apoptosis and autophagy in adenoid cystic carcinoma cells.
复制标题
DOI：
10.1111/odi.12305
复制
发表时间：
2015-05
期刊：
影响因子：
3.8
通讯作者：
Li-zhong Liang;Bin Ma;Yu-Jie Liang;Hai-chao Liu;Tong-Han Zhang;G. Zheng;Yu-xiong Su;Yu-xiong Su;G. Liao
中科院分区：
文献类型：
--
作者：
Li-zhong Liang;Bin Ma;Yu-Jie Liang;Hai-chao Liu;Tong-Han Zhang;G. Zheng;Yu-xiong Su;Yu-xiong Su;G. Liao
腺样囊性癌（Adenoid cystic carcinoma，ACC）是最常见的涎腺肿瘤之一。腺样囊性癌因其高的远处转移率和对化疗或分子治疗不敏感而预后差。本研究旨在探讨Obatoclax对腺样囊性癌细胞的作用及其机制。方法采用Western blot、透射电镜及pEGFP-LC 3质粒转染等方法检测Obatoclax对ACC细胞自噬的影响。3-MA和针对Beclin 1和ATG 5的RNA干扰用于抑制自噬。采用Western blot和Hochest 33342染色检测细胞凋亡。MTT法检测细胞活力。结果Obatoclax诱导腺样囊性癌细胞发生细胞保护性自噬，自噬依赖于ATG 5，部分依赖于Beclin 1。此外，抑制Obatoclax诱导的自噬促进细胞凋亡。Beclin 1或ATG 5的下调通过抑制自噬和凋亡来减弱Obatoclax的细胞毒性。最后，当细胞凋亡被抑制时，自噬性细胞死亡在用Obatoclax处理的腺样囊性癌细胞中开始。结论Beclin 1和ATG 5在Obatoclax诱导的腺样囊性癌细胞自噬和凋亡中起重要调节作用。
OBJECTIVES Adenoid cystic carcinoma (ACC) is one of the most common salivary gland cancers. The prognosis of adenoid cystic carcinoma is poor for its high frequency of distant metastases and insensitivity to chemotherapy or molecular therapies. This study investigated the effect of Obatoclax on adenoid cystic carcinoma cells and its cytotoxic mechanism. METHODS Western blot, transmission electron microscopy, and pEGFP-LC3 plasmids transfection were carried out to detect autophagy in ACC cells treated with Obatoclax. 3-MA and RNA interference against Beclin 1 and ATG5 were used to inhibit autophagy. Then we used Western blot and Hochest 33342 staining for apoptosis assessment. Finally, cell viability was assessed by MTT assay. RESULTS We found that Obatoclax induced cytoprotective autophagy which depended on ATG5 and partly on Beclin 1 in adenoid cystic carcinoma cells. Furthermore, pharmacologically inhibiting Obatoclax-induced autophagy promoted apoptosis. Downregulation of Beclin 1 or ATG5 attenuated the cytotoxicity of Obatoclax by suppressing both autophagy and apoptosis. Finally, when apoptosis was pharmacologically inhibited, autophagic cell death was initiated in adenoid cystic carcinoma cells treated with Obatoclax. CONCLUSION In summary, Beclin 1 and ATG5 play important roles in regulating both Obatoclax-induced autophagy and apoptosis in adenoid cystic carcinoma.