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BMC Cancer. 2017 Nov 10;17(1):750. doi: 10.1186/s12885-017-3721-7.

MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1.

Author information

1
Instituto do Câncer do Estado de São Paulo, Centro de Investigação Translacional em Oncologia, Laboratório de Oncologia Experimental, Av. Dr. Arnaldo,251, São Paulo, SP, CEP 01246-000, Brazil.
2
The University of Texas Health Science Center at San Antonio, Children's Cancer Research Institute, 7703 Floyd Curl Drive, San Antonio, TX, 78229-390, USA.
3
Instituto Hermes Pardini, Setor de Pesquisa e Desenvolvimento, Av das Nações, 2448, Distrito Industrial, Vespasiano, MG, CEP 33200-000, Brazil.
4
Instituto Sírio-Libanês de Ensino e Pesquisa, Centro de Oncologia Molecular, Rua Prof. Daher Cutait, 69, São Paulo, SP, CEP 01308-060, Brazil.
5
The University of Texas Health Science Center at San Antonio, Children's Cancer Research Institute, 7703 Floyd Curl Drive, San Antonio, TX, 78229-390, USA. penalva@uthscsa.edu.

Abstract

BACKGROUND:

Melanoma is the most lethal type of skin cancer. Since chemoresistance is a significant barrier, identification of regulators affecting chemosensitivity is necessary in order to create new forms of intervention. Prohibitin 1 (PHB1) can act as anti-apoptotic or tumor suppressor molecule, depending on its subcellular localization. Our recent data shown that accumulation of PHB1 protects melanoma cells from chemotherapy-induced cell death. Lacking of post-transcriptional regulation of PHB1 could explain this accumulation. Interestingly, most of melanoma patients have down-regulation of microRNA-195. Here, we investigate the role of miR-195, its impact on PHB1 expression, and on chemosensitivity in melanoma cells.

METHODS:

TCGA-RNAseq data obtained from 341 melanoma patient samples as well as a panel of melanoma cell lines were used in an expression correlation analysis between PHB1 and predicted miRNAs. miR-195 impact on PHB1 mRNA and protein levels and relevance of this regulation were investigated in UACC-62 and SK-MEL-5 melanoma lines by RT-qPCR and western blot, luciferase reporter and genetic rescue experiments. Cell proliferation, cell-cycle analysis and caspase 3/7 assay were performed to investigate the potential action of miR-195 as chemosensitizer in melanoma cells treated with cisplatin and temozolomide.

RESULTS:

Analysis of the TCGA-RNAseq revealed a significant negative correlation (Pearson) between miR-195 and PHB1 expression. Moreover, RT-qPCR data showed that miR-195 is down-regulated while PHB1 is up-regulated in a collection of melanoma cells. We demonstrated that miR-195 regulates PHB1 directly by RT-qPCR and western blot in melanoma cells and luciferase assays. To establish PHB1 as a relevant target of miR-195, we conducted rescue experiments in which we showed that PHB1 transgenic expression could antagonize the suppressive effect miR-195 on the proliferation of melanoma cells. Finally, transfection experiments combined with drug treatments performed in the UACC-62 and SK-MEL-5 melanoma cells corroborated miR-195 as potential anti-proliferative agent, with potential impact in sensitization of melanoma cell death.

CONCLUSIONS:

This study support the role of miR-195 as anti-proliferative miRNA via targeting of PHB1 in melanoma cells.

KEYWORDS:

Cisplatin; Melanoma; Prohibitin 1; Temozolomide; Vemurafenib; microRNA-195

PMID:
29126391
PMCID:
PMC5681823
DOI:
10.1186/s12885-017-3721-7
[Indexed for MEDLINE]
Free PMC Article

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