Cobalt chloride induces neurite outgrowth in rat pheochromocytoma PC-12 cells through regulation of endothelin-2/vasoactive intestinal contractor.
We investigated whether endothelin-2/vasoactive intestinal contractor (ET-2/VIC) gene expression, upregulated by hypoxia in cancer cells, was associated with differentiation in neuronal cells. RT-PCR analysis, morphological observations, and immunostaining revealed that CoCl2, a hypoxic mimetic agent, at 200 microM increased expression of the ET-2/VIC gene, decreased expression of the ET-1 gene, and induced neurite outgrowth in PC-12 rat pheochromocytoma cells. These effects induced by 200 microM CoCl2 were completely inhibited by the antioxidant N-acetyl cysteine at 20 mM. In addition, CoCl2 increased the level of intracellular reactive oxygen species (ROS) at an early stage. Furthermore, interleukin (IL)-6 gene expression was upregulated upon the differentiation induced by CoCl2. These results suggest that expression of ET-2/VIC and ET-1 mediated by ROS may be associated with neuronal differentiation through the regulation of IL-6. When the cells were treated with 500 microM CoCl2 for 24 hr, however, ET-2/VIC gene expression disappeared, IL-6 gene expression was downregulated, and necrosis was subsequently induced in the PC-12 cells.
Kotake-Nara
Eiichi
E
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan.
Takizawa
Satoshi
S
Quan
Jiexia
J
Wang
Hongyu
H
Saida
Kaname
K
D000818:Animals; D016587:Antimutagenic Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D003035:Cobalt / Q000494:pharmacology*; D019332:Endothelin-1 / Q000235:genetics / Q000378:metabolism; D019333:Endothelin-2 / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D036341:Intercellular Signaling Peptides and Proteins; D015850:Interleukin-6 / Q000235:genetics; D016501:Neurites / Q000187:drug effects* / Q000502:physiology; D016716:PC12 Cells; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
D016587:Antimutagenic Agents; D000975:Antioxidants; D019332:Endothelin-1; D019333:Endothelin-2; D036341:Intercellular Signaling Peptides and Proteins; D015850:Interleukin-6; D010455:Peptides; D017382:Reactive Oxygen Species; C059568:vasoactive intestinal constrictor; D003035:Cobalt; C018021:cobaltous chloride