MicroRNA-dependent regulation of cKit in cutaneous melanoma

doi:10.1016/j.bbrc.2008.12.152

Biochemical and Biophysical Research Communications

Volume 379, Issue 3, 13 February 2009, Pages 790-794

https://doi.org/10.1016/j.bbrc.2008.12.152 Get rights and content

Abstract

Loss of cKit receptor in cutaneous melanomas was attributed to the down-regulation of AP2 transcription factor. Our analysis of 27 melanoma cell lines showed no correlation between AP2 and c-kit expression. Suggesting a post-transcriptional mechanism of cKit down-modulation, we performed genome-wide microRNA (miRNA) expression profiling and found that several miRNA species are commonly up-regulated in melanomas. Among them was mir-221, which can directly interact with c-kit 3′UTR and inhibit cKit protein translation. Observed inverse correlation of the c-kit and mir-221 expression in various melanocytic cells pointed to its involvement in regulation of cKit in melanoma. Moreover, a series of functional assays demonstrated that mir-221 could directly inhibit cKit, p27^Kip1 and, possibly, other pivotal proteins in melanoma. Collectively, the studies presented here indicate that mir-221 could be a novel therapeutic target for the treatment of cutaneous melanoma. They also suggest that regulation of expression and functional activity of identified up-regulated miRNAs should be further studied in the context of malignant melanoma.

Section snippets

Materials and methods

Cell lines. WM and Lu melanoma cell lines were provided by Dr. M. Herlyn (Wistar Institute, Philadelphia, PA). Primary human melanocytes were provided by Drs. R.E. Boissy and Z. Abdel-Malek (University of Cincinnati, Cincinnati, OH). Melanoma cells were cultured as described at www.wistar.org/herlyn/resources_intro.htm. Primary melanocytes were cultured in melanocyte-specific media with TPA-free supplements (Invitrogen, Carlsbad, CA).

DNA microarray. Analysis of AP2 α and γ isoform expression in

AP2 expression in melanocytic cells

In human melanocytic cells, AP2 transcription factor was shown to be directly involved in regulation of c-kit gene transcription. As only limited number of melanomas were previously tested for the expression of the AP2 α, here we examined the expression of structurally and functionally similar α and γ isoforms of the AP2 in six lines of normal, two lines of vitiligo melanocytes, and 27 lines of melanoma cell isolated from tumors on different stages of tumor progression (Fig. 1A). Based on

Acknowledgments

We thank Drs. M. Herlyn, R.E. Boissy and Z. Abdel-Malek for providing us with various melanocytic cells and for helpful discussions. This research was supported by the Jefferson Medical College to V.A.

References (17)

N. Carpino et al.
p62(dok): a constitutively tyrosine-phosphorylated, GAP-associated protein in chronic myelogenous leukemia progenitor cells
Cell
(1997)
V. Alexeev et al.
Distinctive role of the cKit receptor tyrosine kinase signaling in mammalian melanocytes
J. Invest. Dermatol.
(2006)
S. Galardi et al.
miR-221 and miR-222 expression affects the proliferation potential of human prostate carcinoma cell lines by targeting p27Kip1
J. Biol. Chem.
(2007)
J. Lennartsson et al.
Phosphorylation of Shc by Src family kinases is necessary for stem cell factor receptor/c-kit mediated activation of the Ras/MAP kinase pathway and c-fos induction
Oncogene
(1999)
I. Timokhina et al.
Kit signaling through PI 3-kinase and Src kinase pathways: an essential role for Rac1 and JNK activation in mast cell proliferation
EMBO J.
(1998)
W.W. An et al.
Mitogen-activated protein kinase-dependent apoptosis in norcan-tharidin-treated A375-S2 cells is proceeded by the activation of protein kinase C
Chin. Med. J. (Engl.)
(2005)
J.A. Curtin et al.
Somatic activation of KIT in distinct subtypes of melanoma
J. Clin. Oncol.
(2006)
J.L. Maldonado et al.
Determinants of BRAF mutations in primary melanomas
J. Natl. Cancer Inst.
(2003)

There are more references available in the full text version of this article.

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MicroRNA-dependent regulation of cKit in cutaneous melanoma

Abstract

Section snippets

Materials and methods

AP2 expression in melanocytic cells

Acknowledgments

Cell

J. Invest. Dermatol.

J. Biol. Chem.

Phosphorylation of Shc by Src family kinases is necessary for stem cell factor receptor/c-kit mediated activation of the Ras/MAP kinase pathway and c-fos induction

Oncogene

Kit signaling through PI 3-kinase and Src kinase pathways: an essential role for Rac1 and JNK activation in mast cell proliferation

EMBO J.

Mitogen-activated protein kinase-dependent apoptosis in norcan-tharidin-treated A375-S2 cells is proceeded by the activation of protein kinase C

Chin. Med. J. (Engl.)

Somatic activation of KIT in distinct subtypes of melanoma

J. Clin. Oncol.

Determinants of BRAF mutations in primary melanomas

J. Natl. Cancer Inst.