Overexpression of the miR-34 family suppresses invasive growth of malignant melanoma with the wild-type p53 gene

Yamazaki,Hitoshi; Chijiwa,Tsuyoshi; Inoue,Yoshimasa; Abe,Yoshiyuki; Suemizu,Hiroshi; Kawai,Kenji; Wakui,Masatoshi; Furukawa,Daisuke; Mukai,Masaya; Kuwao,Sadahito; Saegusa,Makoto; Nakamura,Masato

doi:10.3892/etm.2012.497

Overexpression of the miR-34 family suppresses invasive growth of malignant melanoma with the wild-type p53 gene

Authors:
- Hitoshi Yamazaki
- Tsuyoshi Chijiwa
- Yoshimasa Inoue
- Yoshiyuki Abe
- Hiroshi Suemizu
- Kenji Kawai
- Masatoshi Wakui
- Daisuke Furukawa
- Masaya Mukai
- Sadahito Kuwao
- Makoto Saegusa
- Masato Nakamura
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Affiliations: Department of Pathology, Kitasato University School of Medicine, Kanagawa, Japan, Japan Self Defense Force Hospital Naha, Okinawa, Japan, Department of Surgery, Tokai University Hachioji Hospital, Tokyo, Japan, Tokorozawa PET Diagnostic Imaging Clinic, Saitama, Japan, Central Institute for Experimental Animals, Kanagawa, Japan, Department of Laboratory Medicine, Keio University School of Medicine, Tokyo, Japan, Department of Surgery, Tokai University School of Medicine, Kanagawa, Japan, Department of Pathology and Cytology, Higashi-Yamato Hospital, Tokyo, Japan, Regenerative Medicine and Pathology, Tokai University School of Medicine, Kanagawa, Japan
Published online on: February 28, 2012 https://doi.org/10.3892/etm.2012.497
Pages: 793-796

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Abstract

Malignant melanoma is the most aggressive neoplasm, with severe metastatic potential. microRNAs represent a class of endogenously expressed, small non-coding RNAs that regulate gene expression. As a consequence, the translation of these mRNAs is inhibited or they are destabilized resulting in downregulation of the encoded protein. The microRNA-34 (miR-34) family, which comprises three processed microRNAs (miR-34a/b/c) was identified as the mediator of tumor suppression by p53. Many reports suggest that the miR-34s contribute to the inhibition of invasion or metastasis in various tumor types. In this study, we evaluated the expression of the miR-34 family in four human melanoma cell lines (A375, G361, C32TG and SK-MEL-24) which have the wild-type p53 gene using real-time reverse transcription PCR. We also examined their generative and invasive characteristics using the cell proliferation assay and the invasion/migration assay, respectively. All four melanoma cell lines showed significant expression of miR-34s – A375: miR-34a 0.6176, miR-34b 0.7625, miR-34c 0.7877; G361: 7.6424, 16.4127, 22.0332; C32TG: 2.1630, 2.1091, 8.4425; SK-MEL‑24: 0.3621, 2.5659, 8.5907. The cell doubling times of these cell lines in h:min were as follows: A375 23:23, G361 68:24, C32TG 47:22 and SK-MEL-24 67:03. The in vitro generation times of G361 and SK-MEL-24, which showed increased expression of miR-34c, were significantly shorter than A375 with decreased expression of miR-34c (p=0.0063, ANOVA). Invasion (%) of the four cell lines was as follows: A375 44.0%, G361 22.4%, C32TG 13.8% and SK-MEL-24 45.0%. In vitro invasiveness of G361 and C32TG, which showed increased expression of miR-34a, was significantly suppressed (p=0.005, ANOVA). These results suggest that overexpression of miR-34a and c suppresses invasive and generative potentials, respectively, in human malignant melanoma.

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