論文 - 詳細
| RRC ID | 43824 |
|---|---|
| 著者 | Villareal MO, Han J, Ikuta K, Isoda H. |
| タイトル | Mechanism of Mitf inhibition and morphological differentiation effects of hirsein A on B16 melanoma cells revealed by DNA microarray. |
| ジャーナル | J Dermatol Sci |
| Abstract |
BACKGROUND:We have previously reported that hirsein A inhibits melanogenesis in B16 melanoma cells by downregulating the Mitf gene expression. OBJECTIVE:In this study, microarray was employed to determine the transcriptional response of B16 cells to hirsein A (HA) treatment and to find out the mechanism underlying Mitf downregulation. METHODS:DNA microarray, spotted with 265 genes for melanogenesis and signal transduction, was performed using the total RNA isolated from B16 cells treated with HA. Validation of the results was done using real-time PCR. In addition, real-time PCR using primers for Mda-7 gene and F-actin staining were performed. Transfection experiments were performed to knockdown the expression of the Mc1r gene to evaluate its role in the cell morphological change observed. RESULTS:As expected, the expressions of the Mitf-regulated melanosome transport genes and the Mc1r gene were downregulated. Furthermore, the expressions of the MAPK pathway intermediates were either up- or downregulated. Genes associated with cell differentiation, such as Gadd45b, were upregulated and prompted us to determine the expression of the Il-24 (Mda-7) gene using real-time PCR. There was an increase in the Mda-7 mRNA expression in B16 and HMV-II melanoma cells, and in human melanocytes. To better visualize the cell morphology, F-actin staining was performed and the results showed an increase in the dendrite outgrowth in HA-treated cells. Silencing the Mc1r gene did not cause a change in the B16 cell morphology observed in cells treated with HA. CONCLUSION:This study demonstrated that HA downregulates Mitf gene expression by regulating the expressions of the MAPK signaling pathway intermediates. In addition, the inhibited Mc1r gene expression also contributed to the overall Mitf downregulation but does not play a role in the observed change in B16 cell morphology. HA surprisingly can regulate genes associated with differentiating cells (Mda-7) suggesting a role for HA in the melanoma cell differentiation induction. While the exact molecular mechanism by which HA promotes cell differentiation remain to be determined, it is clear that HA can downregulate Mitf expression and promote cell differentiation and has the potential to be used in the development of therapy for melanoma. |
| 巻・号 | 67(1) |
| ページ | 26-36 |
| 公開日 | 2012-7-1 |
| DOI | 10.1016/j.jdermsci.2012.04.005 |
| PII | S0923-1811(12)00137-5 |
| PMID | 22564683 |
| MeSH | Actins / metabolism Animals Antineoplastic Agents, Phytogenic / pharmacology* Cell Differentiation / drug effects* Cell Differentiation / genetics Cell Line, Tumor Cell Shape / drug effects* Cell Shape / genetics Diterpenes / pharmacology* Gene Expression Profiling / methods* Gene Expression Regulation, Neoplastic / drug effects Interleukins / genetics Interleukins / metabolism MAP Kinase Signaling System / drug effects Melanins / biosynthesis Melanoma, Experimental / genetics Melanoma, Experimental / metabolism* Melanoma, Experimental / pathology Mice Microphthalmia-Associated Transcription Factor / antagonists & inhibitors* Microphthalmia-Associated Transcription Factor / metabolism Oligonucleotide Array Sequence Analysis* RNA Interference Real-Time Polymerase Chain Reaction Receptor, Melanocortin, Type 1 / genetics Receptor, Melanocortin, Type 1 / metabolism Reproducibility of Results Transcription, Genetic / drug effects Transfection |
| IF | 3.681 |
| 引用数 | 6 |
| WOS 分野 | DERMATOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | B16 melanoma(RCB1283) |