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

Author(s)

Ramos Cirilo, Priscila Daniele ^{[1, 2, 3]} ; de Sousa Andrade, Luciana Nogueira ^[2] ; Silva Correa, Bruna Renata ^{[1, 4]} ; Qiao, Mei ^[1] ; Furuya, Tatiane Katsue ^[2] ; Chammas, Roger ^[2] ; Ferraz Penalva, Luiz Otavio ^[1]

Affiliation

^[1] Univ Texas Hlth Sci Ctr San Antonio, Childrens Canc Res Inst, 7703 Floyd Curl Dr, San Antonio, TX 78229 - USA

^[2] Inst Canc Estado Sao Paulo, Ctr Invest Translac Oncol, Lab Oncol Expt, Ave Dr Arnaldo 251, BR-01246000 Sao Paulo, SP - Brazil

^[3] Inst Hermes Pardini, Setor Pesquisa & Desenvolvimento, Ave Nacoes 2448, BR-33200000 Vespasiano, MG - Brazil

^[4] Inst Sirio Libanes Ensino & Pesquisa, Ctr Oncol Mol, Rua Prof Daher Cutait 69, BR-01308060 Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: BMC CANCER; v. 17, NOV 10 2017.
• Web of Science Citations: 12

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. (AU)

• FAPESP's process: 98/14247-6 - Center for Research on Cell-Based Therapy
• Grantee: Marco Antonio Zago
• Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 13/11721-0 - Post-transcriptional regulation of PHB gene by RNA-binding proteins and microRNAs in human melanoma
• Grantee: Priscila Daniele Ramos Cirilo
• Support type: Scholarships abroad - Research Internship - Post-doctor
• FAPESP's process: 13/25483-4 - Identification and functional analysis of RNA binding proteins associated with the development of glioblastoma multiform
• Grantee: Bruna Renata Silva Corrêa
• Support type: Scholarships abroad - Research Internship - Doctorate