Abstract
Many studies confirm that circular RNA (circRNA) plays an important regulatory role in the malignant progression of cancer, including melanoma. However, the role of a novel circRNA, circ_0079593, in melanoma is unclear. The expression levels of circ_0079593 and miR-516b were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was measured by cell counting kit-8 (CCK-8) assay, and cell migration and invasion were evaluated using transwell assay. Meanwhile, western blot (WB) analysis was employed to determine the levels of proliferation and metastasis-related proteins, as well as metabotropic glutamate receptor 3 (GRM3) protein. Furthermore, cell apoptosis was tested by detecting the cell apoptosis rate and Caspase-3 activity. The glucose consumption and lactate production of cells were measured to evaluate cell glucose metabolism. Moreover, dual-luciferase reporter assay and biotin-labeled RNA pull-down assay were used to confirm the interaction between miR-516b and circ_0079593 or GRM3. In addition, mice xenograft models were constructed to explore the effect of circ_0079593 on melanoma tumor growth in vivo. Our results discovered that circ_0079593 was highly expressed in melanoma, and its silencing suppressed melanoma cell proliferation, migration, invasion, glucose metabolism and promoted apoptosis. Moreover, we found that circ_0079593 could serve as a sponge of miR-516b, and miR-516b could target GRM3 in melanoma. The rescue experiments revealed that both miR-516b inhibitor and GRM3 overexpression could reverse the inhibition effect of circ_0079593 knockdown on melanoma progression. Additionally, in vivo experiments also revealed that circ_0079593 interference suppressed melanoma tumor growth. Our study concluded that circ_0079593 accelerated melanoma progression via upregulating GRM3 by sponging miR-516b, which suggested that circ_0079593 had the potential to be a new therapeutic biomarker for melanoma.
Similar content being viewed by others
Abbreviations
- qRT-PCR:
-
Quantitative real-time polymerase chain reaction
- CCK-8:
-
Cell counting kit-8
- GRM3:
-
Metabotropic glutamate receptor 3
References
Bagnoni G, Fidanzi C, D'Erme AM, Viacava P, Leoni M, Strambi S, Calani C, Bertocchini A, Morganti R, Spinelli C (2019) Melanoma in children, adolescents and young adults: anatomo-clinical features and prognostic study on 426 cases. Pediatr Surg Int 35(1):159–165
Man W, Wang G (2019) Incidence, outcomes and predictors of primary central nervous system melanoma: a SEER-based study. World Neurosurg 129:e782–e790
Arguello-Guerra L, Vargas-Chandomid E, Diaz-Gonzalez JM, Mendez-Flores S, Ruelas-Villavicencio A, Dominguez-Cherit J (2019) Incidence of complications in dermatological surgery of melanoma and non-melanoma skin cancer in patients with multiple comorbidity and/or antiplatelet-anticoagulants. Five year experience in our Hospital. Cir Cir 86(1):15–23
Tromme I, Sacre L, Hammouch F, Richez P, Degryse JM, Speybroeck N (2018) Melanoma diagnosis: predictive value of macroscopic changes observed by the patient. Melanoma Res 28(6):611–617
Brauner EV, Loft S, Sorensen M, Jensen A, Andersen CE, Ulbak K, Hertel O, Pedersen C, Tjonneland A, Kruger Kjaer S, Raaschou-Nielsen O (2015) Residential radon exposure and skin cancer incidence in a prospective danish cohort. PLoS ONE 10(8):e0135642
Soura E, Eliades PJ, Shannon K, Stratigos AJ, Tsao H (2016) Hereditary melanoma: update on syndromes and management: genetics of familial atypical multiple mole melanoma syndrome. J Am Acad Dermatol 74(3):395–407 quiz 408-310
Hadi I, Roengvoraphoj O, Bodensohn R, Hofmaier J, Niyazi M, Belka C, Nachbichler SB (2020) Stereotactic radiosurgery combined with targeted/ immunotherapy in patients with melanoma brain metastasis. Radiat Oncol 15(1):37
Zhu Z, Liu W, Gotlieb V (2016) The rapidly evolving therapies for advanced melanoma—Towards immunotherapy, molecular targeted therapy, and beyond. Crit Rev Oncol Hematol 99:91–99
Hsiao KY, Sun HS, Tsai SJ (2017) Circular RNA—new member of noncoding RNA with novel functions. Exp Biol Med (Maywood) 242(11):1136–1141
Lei B, Tian Z, Fan W, Ni B (2019) Circular RNA: a novel biomarker and therapeutic target for human cancers. Int J Med Sci 16(2):292–301
Li R, Jiang J, Shi H, Qian H, Zhang X, Xu W (2019) CircRNA: a rising star in gastric cancer. Cell Mol Life Sci 77:1991
Shabaninejad Z, Vafadar A, Movahedpour A, Ghasemi Y, Namdar A, Fathizadeh H, Pourhanifeh MH, Savardashtaki A, Mirzaei H (2019) Circular RNAs in cancer: new insights into functions and implications in ovarian cancer. J Ovarian Res 12(1):84
Tu FL, Guo XQ, Wu HX, He ZY, Wang F, Sun AJ, Dai XD (2020) Circ-0001313/miRNA-510-5p/AKT2 axis promotes the development and progression of colon cancer. Am J Transl Res 12(1):281–291
Yang W, Xie T (2020) Hsa_circ_CSPP1/MiR-361–5p/ITGB1 regulates proliferation and migration of cervical cancer (CC) by modulating the PI3K-Akt signaling pathway. Reprod Sci 27:132
Chen Z, Chen J, Wa Q, He M, Wang X, Zhou J, Cen Y (2020) Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR-429/tribbles homolog 2 axis and Wnt/beta-catenin pathway. Life Sci 243:117323
Zou Y, Wang SS, Wang J, Su HL, Xu JH (2019) CircRNA_0016418 expedites the progression of human skin melanoma via miR-625/YY1 axis. Eur Rev Med Pharmacol Sci 23(24):10918–10930
Bian D, Wu Y, Song G (2018) Novel circular RNA, hsa_circ_0025039 promotes cell growth, invasion and glucose metabolism in malignant melanoma via the miR-198/CDK4 axis. Biomed Pharmacother 108:165–176
Zhang Y, Fu X, Qi Y, Gao Q (2019) A study of the clinical characteristics and prognosis of advanced mucosal and cutaneous melanoma in a Chinese population. Immunotherapy 11(2):91–99
Qu Y, Zhu J, Liu J, Qi L (2019) Circular RNA circ_0079593 indicates a poor prognosis and facilitates cell growth and invasion by sponging miR-182 and miR-433 in glioma. J Cell Biochem 120(10):18005–18013
Pu M, Chen J, Tao Z, Miao L, Qi X, Wang Y, Ren J (2019) Regulatory network of miRNA on its target: coordination between transcriptional and post-transcriptional regulation of gene expression. Cell Mol Life Sci 76(3):441–451
Lorusso C, De Summa S, Pinto R, Danza K, Tommasi S (2020) miRNAs as key players in the management of cutaneous melanoma. Cells 9(2):415
Zhao Y, Wang Y, Xing G (2018) miR-516b functions as a tumor suppressor by directly modulating CCNG1 expression in esophageal squamous cell carcinoma. Biomed Pharmacother 106:1650–1660
Zhu J, Zhang Y, Yang X, Jin L (2017) Clinical significance and tumor-suppressive function of miR-516b in nonsmall cell lung cancer. Cancer Biother Radiopharm 32(4):115–123
Fang G, Ye BL, Hu BR, Ruan XJ, Shi YX (2018) CircRNA_100290 promotes colorectal cancer progression through miR-516b-induced downregulation of FZD4 expression and Wnt/beta-catenin signaling. Biochem Biophys Res Commun 504(1):184–189
Hanniford D, Segura MF, Zhong J, Philips E, Jirau-Serrano X, Darvishian F, Berman RS, Shapiro RL, Pavlick AC, Brown B, Osman I, Hernando E (2015) Identification of metastasis-suppressive microRNAs in primary melanoma. J Natl Cancer Inst 107(3):494
Saini SM, Mancuso SG, Mostaid MS, Liu C, Pantelis C, Everall IP, Bousman CA (2017) Meta-analysis supports GWAS-implicated link between GRM3 and schizophrenia risk. Transl Psychiatry 7(8):e1196
Yi H, Geng L, Black A, Talmon G, Berim L, Wang J (2017) The miR-487b-3p/GRM3/TGFbeta signaling axis is an important regulator of colon cancer tumorigenesis. Oncogene 36(24):3477–3489
Chen A, Wang L, Li BY, Sherman J, Ryu JE, Hamamura K, Liu Y, Nakshatri H, Yokota H (2017) Reduction in migratory phenotype in a metastasized breast cancer cell line via downregulation of S100A4 and GRM3. Sci Rep 7(1):3459
Prickett TD, Wei X, Cardenas-Navia I, Teer JK, Lin JC, Walia V, Gartner J, Jiang J, Cherukuri PF, Molinolo A, Davies MA, Gershenwald JE, Stemke-Hale K, Rosenberg SA, Margulies EH, Samuels Y (2011) Exon capture analysis of G protein-coupled receptors identifies activating mutations in GRM3 in melanoma. Nat Genet 43(11):1119–1126
Neto A, Ceol CJ (2018) Melanoma-associated GRM3 variants dysregulate melanosome trafficking and cAMP signaling. Pigment Cell Melanoma Res 31(1):115–119
Funding
Shanghai Municipal Commission of Health and Family Planning (Grant No. 20174Y0029).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no financial conflicts of interest and non-financial conflict of interest in the manuscript.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Lu, J., Li, Y. Circ_0079593 facilitates proliferation, metastasis, glucose metabolism and inhibits apoptosis in melanoma by regulating the miR-516b/GRM3 axis. Mol Cell Biochem 475, 227–237 (2020). https://doi.org/10.1007/s11010-020-03875-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-020-03875-8