Negative regulation of endogenous protein kinase Cα on the dynamic change of carbachol-induced intracellular calcium response in different melanoma cells
✍ Scribed by Huan Wang; Yao-Qing Yu; Wen-Jun Liao; Zong-Ren Wang; Ya-Jie Lv; Yan-Guo Zhang; Tian-Wen Gao
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 298 KB
- Volume
- 221
- Category
- Article
- ISSN
- 0021-9541
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✦ Synopsis
Abstract
Regulations of intracellular protein kinase C (PKC) on carbachol (CCh)‐induced intracellular calcium ([Ca^2+^]i) responses were investigated in different stages of melanoma cells. We found that CCh (1 mM) significantly increased [Ca^2+^]i with 6‐, 4‐, 4‐, and 25‐folds intensities in WM793B, 451Lu, SK‐MEL‐5, and A2058 melanoma cells, respectively. Pretreatment of phorbol 12, 13‐dibutyrate (PDBu, 2 µM), an activator of intracellular PKC, significantly suppressed CCh‐induced peak reactions in WM793B, SK‐MEL‐5, and A2058 cells. RT‐PCR data showed that mRNA levels of PKCα were 12‐, 4‐, 6‐, and 0.9‐folds higher in above four melanoma cells. Short interfering RNA (siRNA) targeting to PKCα in WM793B cells enhanced CCh‐induced peak calcium reactions. Present data indicated that CCh‐induced [Ca^2+^]i responses were dynamically changed in different stages of melanoma progression. Moreover, intracellular PKCα activated by exogenous agonist and expressed through endogenous gene transcription negatively regulated CCh‐induced calcium responses. The functional analysis on the relationship between CCh‐induced calcium response and endogenous PKCα expression might be helpful to predict the development of melanoma. J. Cell. Physiol. 221: 276–282, 2009. © 2009 Wiley‐Liss, Inc.