miR-193a: potential therapeutic agent and biomarker in cutaneous melanoma Background. MicroRNAs (miRNAs) are short non-coding RNAs regulating gene expression in many biological processes, including proliferation, apoptosis, and differentiation [1]. The relevant role played by miRNAs in cancer, as in other diseases, make them possible new drugs or drug targets as well as diagnostic and prognostic disease biomarkers. MiR-193a has been observed to behave as tumour suppressor in several cancers [2-4] and as oncogene in others [5-6]. In melanoma, the levels of miR-193a was found under-expressed in BRAF-mutated vs wild type cells [7] and in plasma of melanoma patients compared to healthy controls [8]. These evidences suggest a possible oncosuppressor and biomarker function of this miRNA in melanoma. Aim. In the present study, the effects of miR-193a ectopic over-expression in melanoma cells was investigated in order to elucidate its pharmacological influence on melanoma cell proliferation, metastasis and epithelial to mesenchymal transition. Moreover, miR-193-a expression in plasma exosomes derived from metastatic melanoma patients and healthy subjects was evaluated to confirm its role as biomarker. Materials and methods: Commercially available miR-193a-3p and miR-193a-5p, alone or in association, were transfected with lipofectamine in three melanoma cell lines with different BRAF mutation status (A375, 501Mel and MeWo). Transfection with a siRNA scramble was used as control. Cell viability, via Neutral Red assay, pAkt, pErk and Vimentin protein levels, via Western Blotting, were evaluated after exposure to the miR-193a mimic. Exosomes from plasma samples of melanoma patients and healthy donors were isolated and mir-193a levels were determined via quantitative real-time PCR using two types of normalization: versus endogenous miR-16 and exogenous C. elegans miR-39. Results. All miR-193a mimic-transfected cell lines showed a significant cell viability decrease after 72, 96 and 120 h. pAkt, pErk and Vimentin levels were significantly decreased in miR- 193a overexpressing cells compared to controls. Moreover, exosome-derived miR-193a levels were significantly reduced in metastatic melanoma patients compared to healthy donors with a greater decrease observed in patients carrying the BRAF V600E mutation. Conclusions. Our data suggest that miR-193a represents a potential therapeutic agent reducing melanoma progression, independently from BRAF mutation, and merits further evaluation in an in vivo model. Moreover, our results confirm the biomarker role of this circulating miRNA able to discriminate BRAF V600E mutated from non-mutated melanomas. References [1] Tuna M, Machado AS, Calin GA. Genetic and epigenetic alterations of microRNAs and implications for human cancers and other diseases. Genes Chromosomes Cancer 2016; 55(3): 193-214. [2] Yu T, Li J, Yan M, et al. MicroRNA-193a-3p and -5p suppress the metastasis of human non-small-cell lung cancer by downregulating the ERBB4/PIK3R3/mTOR/S6K2 signaling pathway. Oncogene. 2015;34(4):413-23. [3] Williams M, Kirschner MB, Cheng YY, et al. miR-193a-3p is a potential tumor suppressor in malignant pleural mesothelioma. Oncotarget. 2015 Sep 15;6(27):23480-95. [4] Jian B, Li Z, Xiao D, et al. Downregulation of microRNA-193-3p inhibits tumor proliferation migration and chemoresistance in human gastric cancer by regulating PTEN gene. Tumour Biol. 2016;37(7):8941-9. [5] Yi Y, Chen J, Jiao C, et al. Upregulated miR-193a-3p as an oncogene in esophageal squamous cell carcinoma regulating cellular proliferation, migration and apoptosis. Oncol Lett. 2016;12(6):4779-4784. [6] Jian B, Li Z, Xiao D, et al. Downregulation of microRNA-193-3p inhibits tumor proliferation migration and chemoresistance in human gastric cancer by regulating PTEN gene. Tumour Biol. 2016;37(7):8941-9. [7] Caramuta S, Egyházi S, Rodolfo M, et al. MicroRNA expression profiles associated with mutational status and survival in malignant melanoma. J Invest Dermatol. 2010;130(8):2062-70. [8] Fogli S, Polini B, Carpi S, et al. Identification of plasma microRNAs as new potential biomarkers with high diagnostic power in human cutaneous melanoma. Tumour Biol. 2017;39(5):1010428317701646.
miR-193a: potential therapeutic agent and circulating biomarker in cutaneous melanoma
carpi sara
Primo
2018-01-01
Abstract
miR-193a: potential therapeutic agent and biomarker in cutaneous melanoma Background. MicroRNAs (miRNAs) are short non-coding RNAs regulating gene expression in many biological processes, including proliferation, apoptosis, and differentiation [1]. The relevant role played by miRNAs in cancer, as in other diseases, make them possible new drugs or drug targets as well as diagnostic and prognostic disease biomarkers. MiR-193a has been observed to behave as tumour suppressor in several cancers [2-4] and as oncogene in others [5-6]. In melanoma, the levels of miR-193a was found under-expressed in BRAF-mutated vs wild type cells [7] and in plasma of melanoma patients compared to healthy controls [8]. These evidences suggest a possible oncosuppressor and biomarker function of this miRNA in melanoma. Aim. In the present study, the effects of miR-193a ectopic over-expression in melanoma cells was investigated in order to elucidate its pharmacological influence on melanoma cell proliferation, metastasis and epithelial to mesenchymal transition. Moreover, miR-193-a expression in plasma exosomes derived from metastatic melanoma patients and healthy subjects was evaluated to confirm its role as biomarker. Materials and methods: Commercially available miR-193a-3p and miR-193a-5p, alone or in association, were transfected with lipofectamine in three melanoma cell lines with different BRAF mutation status (A375, 501Mel and MeWo). Transfection with a siRNA scramble was used as control. Cell viability, via Neutral Red assay, pAkt, pErk and Vimentin protein levels, via Western Blotting, were evaluated after exposure to the miR-193a mimic. Exosomes from plasma samples of melanoma patients and healthy donors were isolated and mir-193a levels were determined via quantitative real-time PCR using two types of normalization: versus endogenous miR-16 and exogenous C. elegans miR-39. Results. All miR-193a mimic-transfected cell lines showed a significant cell viability decrease after 72, 96 and 120 h. pAkt, pErk and Vimentin levels were significantly decreased in miR- 193a overexpressing cells compared to controls. Moreover, exosome-derived miR-193a levels were significantly reduced in metastatic melanoma patients compared to healthy donors with a greater decrease observed in patients carrying the BRAF V600E mutation. Conclusions. Our data suggest that miR-193a represents a potential therapeutic agent reducing melanoma progression, independently from BRAF mutation, and merits further evaluation in an in vivo model. Moreover, our results confirm the biomarker role of this circulating miRNA able to discriminate BRAF V600E mutated from non-mutated melanomas. References [1] Tuna M, Machado AS, Calin GA. Genetic and epigenetic alterations of microRNAs and implications for human cancers and other diseases. Genes Chromosomes Cancer 2016; 55(3): 193-214. [2] Yu T, Li J, Yan M, et al. MicroRNA-193a-3p and -5p suppress the metastasis of human non-small-cell lung cancer by downregulating the ERBB4/PIK3R3/mTOR/S6K2 signaling pathway. Oncogene. 2015;34(4):413-23. [3] Williams M, Kirschner MB, Cheng YY, et al. miR-193a-3p is a potential tumor suppressor in malignant pleural mesothelioma. Oncotarget. 2015 Sep 15;6(27):23480-95. [4] Jian B, Li Z, Xiao D, et al. Downregulation of microRNA-193-3p inhibits tumor proliferation migration and chemoresistance in human gastric cancer by regulating PTEN gene. Tumour Biol. 2016;37(7):8941-9. [5] Yi Y, Chen J, Jiao C, et al. Upregulated miR-193a-3p as an oncogene in esophageal squamous cell carcinoma regulating cellular proliferation, migration and apoptosis. Oncol Lett. 2016;12(6):4779-4784. [6] Jian B, Li Z, Xiao D, et al. Downregulation of microRNA-193-3p inhibits tumor proliferation migration and chemoresistance in human gastric cancer by regulating PTEN gene. Tumour Biol. 2016;37(7):8941-9. [7] Caramuta S, Egyházi S, Rodolfo M, et al. MicroRNA expression profiles associated with mutational status and survival in malignant melanoma. J Invest Dermatol. 2010;130(8):2062-70. [8] Fogli S, Polini B, Carpi S, et al. Identification of plasma microRNAs as new potential biomarkers with high diagnostic power in human cutaneous melanoma. Tumour Biol. 2017;39(5):1010428317701646.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
