Inhibition of Autophagy Strengthens Celastrol-Induced Apoptosis in Human Pancreatic Cancer In Vitro and In Vivo Models
Inhibition of Autophagy Strengthens Celastrol-Induced Apoptosis in Human Pancreatic Cancer In Vitro and In Vivo Models
复制标题
抑制自噬可增强雷公藤红醇诱导的人胰腺癌体外和体内模型细胞凋亡
DOI：
10.2174/1566524014666140414211223
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发表时间：
2014-05-01
影响因子：
2.5
通讯作者：
Hao, J.
中科院分区：
文献类型：
--
作者：
Zhao, X.;Gao, S.;Hao, J.
目的：雷公藤红素是一种醌甲基化三萜类化合物，能诱导胰腺癌细胞凋亡。本研究的目的是确定是否有保护性自噬后雷公藤红素处理胰腺癌细胞和雷公藤红素和3-MA在体外和体内的协同效应。流式细胞仪检测细胞凋亡程度和自噬空泡数量。免疫荧光法用于监测自噬蛋白LC 3-II的定位。免疫印迹法检测LC 3-II、caspase-3、Bax和bcl-2的表达。电镜观察自噬体。在MiaPaCa-2异种移植瘤模型中研究雷公藤红素和3-MA在体内的协同作用。此外，在体外，当用3-MA抑制自噬时，雷公藤红素诱导的凋亡水平升高;在通过饥饿增强自噬后，雷公藤红素诱导的凋亡水平下降。3-MA增强雷公藤红素诱导的细胞凋亡和对肿瘤生长的抑制作用invivo.Conclusions：在胰腺癌中，雷公藤红素治疗增加了自噬水平，以保护癌细胞免于凋亡。自噬抑制剂3-MA可提高雷公藤红素的体内外治疗效果。
Objectives: Celastrol, a quinone methide triterpenoid, could induce apoptosis in pancreatic cancer cells. The purpose of this study is to determine whether there is protective autophagy after celastrol treatment in pancreatic cancer cells and the synergistic effects of celastrol and 3-MA in vitro and in vivo.Methods: The cells viability was measured using MTT assays. Degree of apoptosis and amount of autophagic vacuoles were measured by flow cytometry. Immunofluorescence was adapted to monitor the localization of autophagic protein LC3-II. Expression of LC3-II, cleaved caspase-3, Bax and bcl-2 was detected by immunoblot. Autophagosomes were observed by electron microscopy. The synergistic effect of celastrol and 3-MA in vivo was studied in the MiaPaCa-2 xenograft tumor model.Results: Celastrol increased the level of autophagy in pancreatic cancer cells. Furthermore in vitro, when inhibiting the autophagy with 3-MA, the level of celastrol-induced apoptosis elevated; after upgrading autophagy by starvation, the level of celastrol-induced apoptosis descended. 3-MA enhanced celastrol-induced apoptosis and inhibitory effect on tumor growth in vivo.Conclusions: In pancreatic cancer, celastrol treatment increased the level of autophagy to protect cancer cells against apoptosis. Autophagy inhibitor 3-MA could improve the therapeutic effect of celastrol in vitro and in vivo.