miR-509-3p Suppresses Migration, Invasion, and Epithelial– Mesenchymal Transition in Melanoma Cells by Targeting CollagenTriple Helix Repeat Containing 1

Background: microRNAs (miRNAs) are ubiquitously dysregulated innumerous tumor cell types, including melanoma cells. The anti-tumoreffect of miR-509-3p was widely evaluated in various cancers. Aims: To determine the functional role of miR-509-3p in melanoma. Study Design: Cell culture study. Methods: Expression of miR-509-3p in melanoma cell models wereassessed by qRT-PCR. Cell migration and invasion were analyzed bywound healing and transwell assays, respectively. Expression levels ofbiomarkers of epithelial–mesenchymal transition were determined byWestern blot. Luciferase vectors containing wildtype or mutant miR 509-3p binding site were constructed, and then dual-luciferase reporterassay. Results: Dysregulated miR-509-3p level was found in melanoma cells.Elevated miR-509-3p expression suppressed melanoma cell migration(P < .001) and invasion (P < .001) capacities. Epithelial–mesenchymaltransition of melanoma cells was repressed by miR-509-3p, along withincreased α-catenin/E-cadherin (P < .001) and decreased vimentin/ fibronectin (P < .001). CTHRC1 (collagen triple helix repeat contain ing 1) contained a potential binding site for miR-509-3p, and miR 509-3p decreased protein expression of CTHRC1 in melanoma cells(P < .001). CTHRC1 promoted melanoma cell migration and invasion(P < .001), as well as contributed to epithelial–mesenchymal transition.Increased CTHRC1 expression attenuated miR-509-3p-induced inhibi tion of melanoma cell migration (P < .001), invasion, and epithelial– mesenchymal transition. Conclusion: miR-509-3p suppressed the biological function ofmelanoma cells through negatively regulating CTHRC1, shedding lighton miR-509-3p as a potential candidate for melanoma therapeutics andtreatments.

PDF

___

1. Jang YC, Liang SJ. Vaginal primary malignant melanoma of a Chinese woman: an exceptionally rare yet lethal case report and literature review. Eur J Gynaecol Oncol. 2018;39:327-330.
2. Fraihat A, Alatrash L, Abbasi R, et al. Inhibitory effects of methanol extracts of selected plants on the proliferation of two human melanoma cell lines. Trop J Pharm Res. 2018;17(6):1081-1086. [CrossRef]
3. Segura MF, Belitskaya-Lévy I, Rose AE, et al. Melanoma microRNA signature predicts post-recurrence survival. Clin Cancer Res. 2010;16(5):1577-1586. [CrossRef]
4. Shukla GC, Singh J, Barik S. MicroRNAs: processing, maturation, target recognition and regulatory functions. Mol Cell Pharmacol. 2011;3(3):83-92.
5. Lee YS, Dutta A. MicroRNAs in cancer. Annu Rev Pathol. 2009;4:199-227. [CrossRef]
6. Mione M, Bosserhoff A. MicroRNAs in melanocyte and melanoma biology. Pigment Cell Melanoma Res. 2015;28(3):340-354. [CrossRef]
7. Lorusso C, De Summa S, Pinto R, Danza K, Tommasi S. miRNAs as key players in the management of cutaneous melanoma. Cells. 2020;9(2):415. [CrossRef]
8. Zhai Q, Zhou L, Zhao C, et al. Identification of miR-508-3p and miR-509-3p that are associated with cell invasion and migration and involved in the apoptosis of renal cell carcinoma. Biochem Biophys Res Commun. 2012;419(4):621-626. [CrossRef]
9. Pan Y, Robertson G, Pedersen L, et al. miR-509-3p is clinically significant and strongly attenuates cellular migration and multi-cellular spheroids in ovarian cancer. Oncotarget. 2016;7(18):25930-25948. [CrossRef]
10. Díaz-Martínez M, Benito-Jardón L, Alonso L, et al. miR-204-5p and miR-211-5p contribute to BRAF inhibitor resistance in melanoma. Cancer Res. 2018;78(4):1017- 1030. [CrossRef]
11. Stark MS, Klein K, Weide B, et al. The prognostic and predictive value of melanomarelated microRNAs using tissue and serum: a microRNA expression analysis. EBiomedicine. 2015;2(7):671-680. [CrossRef]
12. Tang L, Dai DL, Su M, et al. Aberrant expression of collagen triple helix repeat containing 1 in human solid cancers. Clin Cancer Res. 2006;12(12):3716-3722. [CrossRef]
13. Jiang N, Cui Y, Liu J, et al. Multidimensional roles of collagen triple helix repeat containing 1 (CTHRC1) in malignant cancers. J Cancer. 2016;7(15):2213-2220. [CrossRef]
14. Luo Z, Zhang X, Zeng W, et al. TRAF6 regulates melanoma invasion and metastasis through ubiquitination of basigin. Oncotarget. 2016;7(6):7179-7192. [CrossRef]
15. Scott KL, Nogueira C, Heffernan TP, et al. Proinvasion metastasis drivers in earlystage melanoma are oncogenes. Cancer Cell. 2011;20(1):92-103. [CrossRef]
16. Mumford SL, Towler BP, Pashler AL, et al. Circulating microRNA biomarkers in melanoma: tools and challenges in personalised medicine. Biomolecules. 2018;8(2):21. [CrossRef]
17. Philippidou D, Schmitt M, Moser D, et al. Signatures of microRNAs and selected microRNA target genes in human melanoma. Cancer Res. 2010;70(10):4163-4173. [CrossRef]
18. Li Y, Li M, Shats I, et al. Glypican 6 is a putative biomarker for metastatic progression of cutaneous melanoma. PLoS ONE. 2019;14(6):e0218067. [CrossRef]
19. Hodorogea A, Calinescu A, Antohe M, et al. Epithelial-mesenchymal transition in skin cancers: a review. Anal Cell Pathol. 2019;2019:3851576. [CrossRef]
20. Song J, Chen X, Zhang L, Song D, Xiong H. MicroRNA-204-3p modulates epithelialmesenchymal transition by targeting paired box gene 2 in human melanoma A-375 cells. Transl Cancer Res. 2019;8(5):2032-2043. [CrossRef]
21. Zhang J, Ma L. MicroRNA control of epithelial-mesenchymal transition and metastasis. Cancer Metastasis Rev. 2012;31(3-4):653-662. [CrossRef]
22. Park EH, Kim S, Jo JY, et al. Collagen triple helix repeat containing-1 promotes pancreatic cancer progression by regulating migration and adhesion of tumor cells. Carcinogenesis. 2013;34(3):694-702. [CrossRef]
23. Eriksson J, Le Joncour V, Nummela P, et al. Gene expression analyses of primary melanomas reveal CTHRC1 as an important player in melanoma progression. Oncotarget. 2016;7(12):15065-15092. [CrossRef]
24. Turkar S, Ramaswamy A, Ostwal V. A step ahead on CTHRC1, and not just reinventing the wheel! Chin Clin Oncol. 2019;8(S1):S17. [CrossRef]
25. Li Y, Fu Y, Gao Y, et al. microRNA-134 inhibits melanoma growth and metastasis by negatively regulating collagen triple helix repeat containing-1 (CTHRC1). Int J Clin Exp Pathol. 2018;11(9):4319-4330.
26. Li Y, Zhang Y, Ma C, et al. Overexpression of CTHRC1 in human melanoma promotes tumorigenesis targeted by miRNA155. Int J Clin Exp Pathol. 2017;10(8):8199-8210.