Sodium arsenite-induced DAPK promoter hypermethylation and autophagy via ERK1/2 phosphorylation in human uroepithelial cells.
Arsenic compounds or arsenicals are well-known toxic and carcinogenic agents. The toxic effects of arsenic that are of most concern to humans are those that occur from chronic, low-level exposure, and are associated with various human malignancies, including skin, lung and bladder cancers. In addition, arsenic could induce cell death, including apoptosis or autophagy in malignant cells. Previously, we have demonstrated that arsenite can induce autophagy and death-associated protein kinase (DAPK) promoter hypermethylation in the SV-40 immortalized human uroepithelial cell line (SV-HUC-1). However, the underlying mechanism of arsenite-induced autophagy is still unclear. In the present study, we demonstrate that arsenite can activate the extracellular signaling-regulated protein kinase 1/2 (ERK1/2) signaling pathway after treatment in SV-HUC-1 cells by using immunocytochemistry and Western blotting. In addition, our results also show an increase of autophagosomes was produced in arsenite-treated SV-HUC-1 cells by using electron microscopy. We found that, by incrementally increasing the dosages, microtubule-associated protein light chain 3B (LC3B) and Beclin-1 are important regulators for the formation of autophagosomes, in a dose-dependent manner. When the cells were pretreated with inhibitors 5-aza-CdR or U0126 for 24h, the effect of arsenite on ERK1/2, LC3B, Beclin-1 and DAPK proteins expression is suppressed. Furthermore, our results support the notion that arsenite can induce the ERK1/2 signaling pathway to stimulate autophagy and DAPK promoter hypermethylation in human uroepithelial SV-HUC-1 cells. These findings may contribute to a better understanding of the carcinogenesis of arsenite.
Huang
Ya-Chun
YC
Kaohsiung Medical University, Taiwan.
Hung
Wen-Chun
WC
Chen
Wan-Tzu
WT
Yu
Hsin-Su
HS
Chai
Chee-Yin
CY
D051017:Apoptosis Regulatory Proteins / Q000235:genetics*; D018053:Arsenites / Q000633:toxicity*; D001343:Autophagy / Q000187:drug effects*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics*; D002460:Cell Line; D019175:DNA Methylation / Q000187:drug effects*; D064550:Death-Associated Protein Kinases; D004789:Enzyme Activation; D006801:Humans; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D017670:Sodium Compounds / Q000633:toxicity*; D019459:Urothelium / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure
D051017:Apoptosis Regulatory Proteins; D018053:Arsenites; D017670:Sodium Compounds; C017947:sodium arsenite; D064550:Death-Associated Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3