Circular RNA circRNA_0082835 promotes progression and lymphatic metastasis of primary melanoma by sponging microRNA miRNA-429

ABSTRACT

To identify how circular RNA circRNA_0082835 impacts melanoma cells and lymphatic metastasis to observe whether it exerts effects through its action mechanism of sponging microRNA miR-429. Clinical baseline information was collected, and clinical samples were used for detection on circRNA_0082835 and EZH2. The expression of circRNA_0082835, EZH2, and miR-429 was detected by quantitative real-time PCR (RT-qPCR). Cell proliferation was tested with cell counting kit-8 (CCK-8). Flow cytometry was applied to examination of cell cycle levels. Cell invasion and migration were observed by transwell and wound healing. The expression of Wnt/β-catenin pathway, cell cycle and epithelial–mesenchymal transition (EMT) marker proteins was analyzed by western blot. Dual-luciferase determined the binding of miR-429 and circ_0082835. As a result, the expression of circRNA_0082835 was increased and that of miR-429 was decreased with the increase in lymphatic metastasis level. CircRNA_0082835 expression was downregulated by circ_0082835 interference, upregulated by EZH2 interference and also downregulated after transfection of both shRNA-circ_0082835 and shRNA-EZH2. Inhibiting circ_0082835 and EZH2 suppressed the proliferation, invasion and migration, regulated the cell cycle levels, inhibited Wnt/β-catenin and attenuated EMT in melanoma cells. Inhibition of circ_0082835 and/or EZH2 elevated miR-429 expression. The binding among miR-429 and circ_0082835 was verified. MiR-429 inhibitor reversed the effect of circ_0082835 interference while having no significant impact on EZH2. In conclusion, circRNA_0082835 sponges miR-429 to affect the anti-tumor effect of miR-429 in primary melanoma and lymphatic metastasis.

Graphical Abtstract

KEYWORDS:

• CircRNA_0082835
• miR-429
• primary melanoma
• EZH2
• Wnt/β-catenin

Acknowledgements

The authors thank all for participating in this study and laboratory assay. We also thank Han Chen MD, Ph.D. for her terrific advice, critical comments, and technical assistance with this manuscript.

Disclosure statement

The author(s) declare that they have no competing interests. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Ethics approval

All procedures performed were in accordance with the declaration of the ethical standards of the institutional research committee and with the 1964 Helsinki 387 Declaration and its later amendments. The ethics committee has approved this study of the First Affiliated Hospital of Nanjing Medical University between December 2018 and December 2020 in our department (No. 2020-SR-563).

Consent to participate

All the patients provided informed consent upon this study.

Consent for publication

Written informed was obtained from the patient for the publication of this study and any accompanying images. A copy of written consent is available for review by the Editor-in-Chief of this journal.

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Authors’ contributions

YTS collect the clinical data, complete the manuscript and translation. ZQH helped to draft the manuscript and participated in the experiments. BLL, CJL, JFL, and JLL supported collecting the data, participated in the experiments and follow-up patients. JT and GY designed and supervised this study also revised the manuscript for important intellectual content and translation. All authors read and approved the final manuscript.

Additional information

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.