Nie Jifeng, Liu Aidong, Tan Qunya, Zhao Kai, Hu Kui, Li Yong, Yan Bin, Zhou Lin
Department of Minimally Invasive Surgery, Integrated Chinese and Western Medicine Hospital of Zhejiang Province, Hangzhou, China.
Department of Pathology, North China University of Science and Technology Affiliated Hospital, Tangshan, China.
Biochem Biophys Res Commun. 2017 Jan 8;482(2):246-252. doi: 10.1016/j.bbrc.2016.11.050. Epub 2016 Nov 12.
AICAR (5-Aminoimidazole-4-carboxamide riboside or acadesine) is an AMP-activated protein kinase (AMPK) agonist, its activity in human gallbladder cancer cells was evaluated here. We show that AICAR provoked significant apoptosis in human gallbladder cancer cell lines (Mz-ChA-1, QBC939 and GBC-SD) and primary gallbladder cancer cells. AICAR-induced cytotoxicity in gallbladder cancer cells appears independent of AMPK activation. Inhibition of AMPK, via AMPKα shRNA knockdown or dominant negative mutation (T172A), failed to rescue GBC-SD cells from AICAR. Further, forced-activation of AMPK, by adding two other AMPK activators (A769662 and Compound 13), or expressing a constitutively-active mutant AMPKα (T172D), didn't induce GBC-SD cell death. Remarkably, AICAR treatment in gallbladder cancer cells induced endoplasmic reticulum (ER) stress activation, the latter was tested by caspase-12 activation, C/EBP homologous protein (CHOP) expression and IRE1/PERK phosphorylation. Contrarily, salubrinal (the ER stress inhibitor), z-ATAD-fmk (the caspase-12 inhibitor) or CHOP shRNAs significantly attenuated AICAR-induced gallbladder cancer cell apoptosis. Together, we conclude that AICAR-induced gallbladder cancer cell apoptosis requires ER stress activation, but is independent of AMPK.
AICAR（5-氨基咪唑-4-甲酰胺核苷或阿卡地新）是一种AMP激活的蛋白激酶（AMPK）激动剂，本文评估了其在人胆囊癌细胞中的活性。我们发现，AICAR可引发人胆囊癌细胞系（Mz-ChA-1、QBC939和GBC-SD）及原发性胆囊癌细胞显著凋亡。AICAR诱导的胆囊癌细胞细胞毒性似乎与AMPK激活无关。通过AMPKα短发夹RNA敲低或显性负性突变（T172A）抑制AMPK，无法使GBC-SD细胞免受AICAR的影响。此外，添加另外两种AMPK激活剂（A769662和化合物13）或表达组成型活性突变体AMPKα（T172D）来强制激活AMPK，并未诱导GBC-SD细胞死亡。值得注意的是，AICAR处理胆囊癌细胞可诱导内质网（ER）应激激活，这通过半胱天冬酶-12激活、C/EBP同源蛋白（CHOP）表达及IRE1/PERK磷酸化来检测。相反，内质网应激抑制剂水杨酰胺、半胱天冬酶-12抑制剂z-ATAD-fmk或CHOP短发夹RNA可显著减弱AICAR诱导的胆囊癌细胞凋亡。我们共同得出结论，AICAR诱导的胆囊癌细胞凋亡需要内质网应激激活，但与AMPK无关。
