Negative regulation of endogenous protein kinase Calpha on the dynamic change of carbachol-induced intracellular calcium response in different melanoma cells.

Full 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 microM), 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 PKCalpha were 12-, 4-, 6-, and 0.9-folds higher in above four melanoma cells. Short interfering RNA (siRNA) targeting to PKCalpha 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 PKCalpha 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 PKCalpha expression might be helpful to predict the development of melanoma.

Author information

Author/s: Wang, Huan (H); Yu, Yao-Qing (YQ); Liao, Wen-Jun (WJ); Wang, Zong-Ren (ZR); Lv, Ya-Jie (YJ); Zhang, Yan-Guo (YG); Gao, Tian-Wen (TW);

Affiliation: Department of Dermatology, Center of Dermatology of Chinese PLA, Xijing Hospital, Fourth Military Medical University, Xi'an, China.

Journal and publication information

Publication Type: Journal Article

Journal: Journal of cellular physiology (J Cell Physiol), published in United States. (Language: eng)

Reference: 2009-Nov; vol 221 (issue 2) : pp 276-82

Dates: Created 2009/08/27; Completed 2009/09/09;

PMID: 19626679, status: MEDLINE (last retrieval date: 9/9/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Adolescent
Calcium Signaling - drug effects
Carbachol - pharmacology
Cell Line, Tumor
Gene Expression Regulation, Neoplastic - drug effects
Gene Knockdown Techniques
Humans
Intracellular Space - drug effects; metabolism
Male

Male
Melanocytes - drug effects; enzymology
Melanoma - enzymology; genetics; pathology
Phorbol 12,13-Dibutyrate - pharmacology
Protein Kinase C-alpha - genetics; metabolism
RNA, Messenger - genetics; metabolism
RNA, Small Interfering - metabolism
Transfection
Wound Healing - drug effects

Associated Chemicals: RNA, Messenger (0) ; RNA, Small Interfering (0) ; Phorbol 12,13-Dibutyrate (37558-16-0) ; Carbachol (51-83-2) ; Protein Kinase C-alpha (EC 2.7.1.37)

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