Autophagy augmented by troglitazone is independent of EGFR transactivation and correlated with AMP-activated protein kinase signaling.
Troglitazone is a synthetic ligand of peroxisome proliferators activated receptor-gamma (PPARgamma) and induces apoptosis in a variety of malignant cells. However, the underlying mechanism of its regulatory role in macroautophagy (hereafter autophagy) remains largely unknown. Using fluorescence and electron microscopy, we observed that autophagosomes could be induced and identified upon troglitazone challenge in both primary and epidermal growth factor receptor (EGFR)-expressed porcine aortic endothelial (PAE) cells. We report here that troglitazone augments AMP-activated protein kinase-alpha (AMPKalpha) phosphorylation, reduces p70S6 kinase phosphorylation and stimulates autophagy that is independent of EGFR expression and transactivation. Troglitazone stimulus reduced neither lysosomal staining nor GFP-LC3 dots of HeLa cells, when the cells pretreated with AG1478, a specific EGFR kinase inhibitor. Furthermore, AG1478 additively enhanced the troglitazone-induced degradation of sequestosome 1 (SQSTM1/p62), which is a selective substrate of autophagy. Inhibition of AMPKalpha activity either by compound C or by RNA interference markedly reduced the accumulation of microtubule-associated protein 1 light chain 3-II (LC3-II), a good indicator of autophagy; whereas blockage of PPARgamma activity by the irreversible antagonist GW9662 or by overexpressing dominate-negative PPARgamma did not affect LC3-II accumulation and AMPK phosphorylation. Taken together, we demonstrate that autophagy promoted via troglitazone is correlated with AMPKalpha activation and independent of PPARgamma activation and EGFR transactivation.
Yan
Jinguo
J
Biological Resource Center, The Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Yang
Huaiyi
H
Wang
Gang
G
Sun
Lei
L
Zhou
Yuguang
Y
Guo
Yinglu
Y
Xi
Zhijun
Z
Jiang
Xuejun
X
D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D001343:Autophagy / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D047495:PPAR gamma / Q000819:agonists / Q000378:metabolism / Q000502:physiology; D010588:Phagosomes / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D045162:Thiazolidinediones / Q000494:pharmacology*; D015533:Transcriptional Activation / Q000187:drug effects; D000077288:Troglitazone; D015854:Up-Regulation / Q000187:drug effects
D002839:Chromans; D047495:PPAR gamma; D045162:Thiazolidinediones; D066246:ErbB Receptors; D055372:AMP-Activated Protein Kinases; D000077288:Troglitazone