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Apigenin inhibits growth of melanoma by suppressing miR-512-3p and promoting the G1 phase of cell cycle involving the p27 Kip1 protein

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Abstract

In the present study, we screened multiple melanoma cell lines for treatment of Apigenin and miRNA expression, also studied the role of miR-512-3p in melanoma. RT-PCR analysis was done for screening miRNA in melanoma cell lines (WM1361B, WM983A, WM1341D, SK-MEL-3, SH-4, SK-MEL-24 and RPMI-7951) compared to normal human epidermal melanocytes. Colony formation assay for cell viability studies, cell cycle by flowcytometry and protein expression by immunoblot analysis. For in vivo analysis tumour xenograft mouse model was created. Immunohistochemistry was done for PCNA positive cells. For expression of miR-512-3p in tumour tissues fluorescence in situ hybridization was done. In silico studies were done by molecular docking studies. The WM1361B and WM983A cell lines showed overexpression of miR-512-3p and increased cell proliferation compared to normal human epidermal melanocytes. Treatment of anti-miR-512-3p to WM1361B and WM983A cells halted cell proliferation and also caused G1-phase arrest. We studied the effect of Apigenin on the expression levels of miR-512-3p and associated molecular targets. Apigenin treatment in WM1361B and WM983A cells showed inhibition in expression of miR-512-3p, arrest of G1 phase of cell cycle, cytotoxicity and revival of p27 Kip1. Apigenin treatment significantly suppressed the growth of WM1361B in tumour induced mice, the activity was associated with decreased levels of miR-512-3p, tumour cell proliferation and increased levels of p27 Kip1 protein. Docking studies confirm potential affinity of Apigenin for p27 Kip1. Apigenin acts as an inhibitor of miR-512-3p by suppressing growth of melanoma both in vitro and in vivo targeting the p27 Kip1 axis.

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All data generated or analysed during this study are included in this article. Further any enquiries can be directed to the corresponding author.

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Acknowledgements

The authors are thankful to the staff and management of Xiangyang Central Hospital for providing the required support.

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The work received no external funding.

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Author notes

  1. Qun Xie and Ruirui Zhang have contributed to this work equally.

Authors and Affiliations

  1. Department of Plastic Surgery, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, No. 136 Jingzhou Road, Xiangcheng, Xiangyang, 441021, Hubei, People’s Republic of China

    Qun Xie, Ruirui Zhang, Dandan Liu, Jing Yang, Qiang Hu, Chao Shan & Xiaohan Li

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  1. Qun Xie
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  2. Ruirui Zhang
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  3. Dandan Liu
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  4. Jing Yang
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Contributions

QX and RZ contributed equally to this work. JY, DL, QH, CS, XL along with QX and RZ designed and performed experiments. All the authors contributed in preparing the manuscript and read the paper before submission.

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Correspondence to Dandan Liu or Jing Yang.

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The animal experiments were approved by the Xiangyang Central Hospital, China.

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Xie, Q., Zhang, R., Liu, D. et al. Apigenin inhibits growth of melanoma by suppressing miR-512-3p and promoting the G1 phase of cell cycle involving the p27 Kip1 protein. Mol Cell Biochem 477, 1569–1582 (2022). https://doi.org/10.1007/s11010-022-04363-x

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