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Regulatory expression of genes related to metastasis by TGF-β and activin A in B16 murine melanoma cells

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Abstract

TGF-β induces epithelial-mesenchymal transition, which occurs during tumor cell invasiveness in pathological state, in limited cells. As a first step to understand the role of TGF-β and the structurally related activin during melanoma metastasis, expression of metastasis-related genes was examined in murine melanoma cells. Treatment with TGF-β1 or activin A down-regulated E-cadherin in B16 cells in a dose-dependent manner. In epithelial cells, TGF-β-induced high mobility group A2 (HMGA2) gene product is suggested to down-regulate E-cadherin through up-regulation of zinc-finger transcription factors Slug and Snail, and basic helix-loop-helix transcription factor Twist. Unlike the regulation in epithelial cells, TGF-β1 treatment rather decreased mRNA expression of HMGA2, indicating a distinct mechanism on TGF-β/activin-induced down-regulation. Transfection of double-stranded interfering RNA (dsRNAi) for activin receptor-like kinase (ALK) type I receptors revealed that ALK5, a prototype of TGF-β receptor, mainly transmits TGF-β signals on the E-cadherin down-regulation at the mRNA level, and that a prototype receptor ALK4 elicited the activin effect. TGF-β/activin potentiated down-regulation of E-cadherin and HMGA2 also in B16 sublines that are susceptible to metastasis. However, the extent of down-regulation tended to be smaller, and less Smad2, a signal mediator for TGF-β/activin, was phosphorylated in response to the ligand, resulting from less expression of type I receptors in the B16 sublines. These results suggest that the receptor expression level determines strength of the signals for TGF-β/activin through phosphorylation of Smad2, which explains pluripotency of the ligand family partly.

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Acknowledgments

We thank Dr. A.F. Parlow for providing recombinant human activin A. This work was supported by Kakenhi from the Japan Society for the Promotion of Science, by grants for Graduate Schools from The Foundation for Japanese Private School Promotion, and by The Science Research Promotion Fund from The Promotion and Mutual Aid Corporation for Private Schools of Japan.

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Authors and Affiliations

  1. Laboratory of Molecular Biology, Azabu University School of Veterinary Medicine, Sagamihara, 229-8501, Japan

    Masaru Murakami & Makiko Suzuki

  2. Laboratory of Veterinary Immunology, Azabu University School of Veterinary Medicine, Sagamihara, 229-8501, Japan

    Yoshii Nishino

  3. Laboratory of Nutrition, Azabu University School of Veterinary Medicine, Sagamihara, 229-8501, Japan

    Masayuki Funaba

  4. Laboratory of Nutritional Science, Kyoto University Graduate School of Agriculture, Kitashirakawa Oiwakecho, Kyoto, 606-8502, Japan

    Masayuki Funaba

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  1. Masaru Murakami
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  2. Makiko Suzuki
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  4. Masayuki Funaba
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Correspondence to Masayuki Funaba.

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Murakami, M., Suzuki, M., Nishino, Y. et al. Regulatory expression of genes related to metastasis by TGF-β and activin A in B16 murine melanoma cells. Mol Biol Rep 37, 1279–1286 (2010). https://doi.org/10.1007/s11033-009-9502-x

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  • DOI: https://doi.org/10.1007/s11033-009-9502-x

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