Title

Copper Induces Apoptosis of Neuroblastoma Cells Via Post-translational Regulation of the Expression of Bcl-2-Family Proteins and the txJ Mouse is a Better Model of Hepatic than Brain Cu Toxicity

Document Type

Journal Article

Publisher

e-Century Publishing

Faculty

Computing, Health and Science

School

Exercise, Biomedical and Health Science, Centre for Alzheimer's Disease

RAS ID

5848

Comments

This article was originally published as: Chan, H., Liu, T., Verdile, G., Glenda, B., Haasl, R., Smith, M., Perry, G., Martins, R. N., & Atwood, C. (2008). Copper Induces Apoptosis of Neuroblastoma Cells Via Post-translational Regulation of the Expression of Bcl-2-family Proteins and the txJ Mouse is a Better Model of Hepatic than Brain Cu Toxicity. International Journal of Clinical and Experimental Medicine, 1(1), 76-88. Original article available here

Abstract

The basic mechanism(s) by which altered Cu homeostasis is toxic to hepatocytes and neurons, the two major cell types affected in copper storage diseases such as Wilson’s disease (WD), remain unclear. Using human M17 neuroblastoma cells as a model to examine Cu toxicity, we found that there was a time- and concentration-dependent induction of neuronal death, such that at 24 h there was a ~50 % reduction in viability with 25 uM Cu-glycine2. Cu-glycine2 (25:50 uM) treatment for 24 h significantly altered the expression of 296 genes, including 8 genes involved with apoptosis (BCL2-associated athanogene 3, BCL2/adenovirus E1B 19kDa interacting protein, caspase 5, regulator of Fas-induced apoptosis, V-jun sarcoma virus 17 oncogene homology, claudin 5, prostaglandin E receptor 3 and protein tyrosine phosphatase, non-receptor type 6). Surprisingly, changes in the expression of more ‘traditional’ apoptotic genes (Bcl-2, Bax, Bak and Bad) did not vary more than 20 %. To test whether the induction of apoptosis in neuroblastoma cells was via posttranslational mechanisms, we measured the protein expression of these apoptotic markers in M17 neuroblastoma cells treated with Cu-glycine2 (0-100 uM) for 24-48 h. Compared with glycine treated cells, Cu-glycine2 reduced Bcl-2 expression by 50 %, but increased Bax and Bak expression by 130% and 400 %, respectively. To assess whether Cu also induced apoptotic cell death in a mouse model of WD, we measured the expression of these apoptotic markers in the liver and brain of mice expressing a ATP7b gene mutation (tx-J mice) at 10 months of age (near the end of their lives when overt liver pathology is displayed). Changes in the liver expression of these apoptotic markers in tx-J mice compared to background mice mirrored those of Cu treated neuroblastoma cells. In contrast, few changes in apoptotic protein expression were detected in the brain between tx-J and background mice, indicating the tx-J mouse is a good model of hepatic, but not brain, Cu toxicity. Our results indicate that Cu-induction of neuronal apoptosis does not require de novo synthesis or degradation of apoptotic genes, and that Cu accumulation in the aged tx-J mouse brain is insufficient to induce apoptosis. (IJCEM710001).