MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1
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 mo...
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doaj-4658ee86154f438c926cdb7a302ae30a2020-11-25T02:17:16ZengBMCBMC Cancer1471-24072017-11-0117111210.1186/s12885-017-3721-7MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1Priscila Daniele Ramos Cirilo0Luciana Nogueira de Sousa Andrade1Bruna Renata Silva Corrêa2Mei Qiao3Tatiane Katsue Furuya4Roger Chammas5Luiz Otavio Ferraz Penalva6Instituto do Câncer do Estado de São Paulo, Centro de Investigação Translacional em Oncologia, Laboratório de Oncologia ExperimentalInstituto do Câncer do Estado de São Paulo, Centro de Investigação Translacional em Oncologia, Laboratório de Oncologia ExperimentalThe University of Texas Health Science Center at San Antonio, Children’s Cancer Research InstituteThe University of Texas Health Science Center at San Antonio, Children’s Cancer Research InstituteInstituto do Câncer do Estado de São Paulo, Centro de Investigação Translacional em Oncologia, Laboratório de Oncologia ExperimentalInstituto do Câncer do Estado de São Paulo, Centro de Investigação Translacional em Oncologia, Laboratório de Oncologia ExperimentalThe University of Texas Health Science Center at San Antonio, Children’s Cancer Research InstituteAbstract 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.http://link.springer.com/article/10.1186/s12885-017-3721-7MelanomamicroRNA-195Prohibitin 1CisplatinTemozolomideVemurafenib |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Priscila Daniele Ramos Cirilo Luciana Nogueira de Sousa Andrade Bruna Renata Silva Corrêa Mei Qiao Tatiane Katsue Furuya Roger Chammas Luiz Otavio Ferraz Penalva |
spellingShingle |
Priscila Daniele Ramos Cirilo Luciana Nogueira de Sousa Andrade Bruna Renata Silva Corrêa Mei Qiao Tatiane Katsue Furuya Roger Chammas Luiz Otavio Ferraz Penalva MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 BMC Cancer Melanoma microRNA-195 Prohibitin 1 Cisplatin Temozolomide Vemurafenib |
author_facet |
Priscila Daniele Ramos Cirilo Luciana Nogueira de Sousa Andrade Bruna Renata Silva Corrêa Mei Qiao Tatiane Katsue Furuya Roger Chammas Luiz Otavio Ferraz Penalva |
author_sort |
Priscila Daniele Ramos Cirilo |
title |
MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 |
title_short |
MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 |
title_full |
MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 |
title_fullStr |
MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 |
title_full_unstemmed |
MicroRNA-195 acts as an anti-proliferative miRNA in human melanoma cells by targeting Prohibitin 1 |
title_sort |
microrna-195 acts as an anti-proliferative mirna in human melanoma cells by targeting prohibitin 1 |
publisher |
BMC |
series |
BMC Cancer |
issn |
1471-2407 |
publishDate |
2017-11-01 |
description |
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. |
topic |
Melanoma microRNA-195 Prohibitin 1 Cisplatin Temozolomide Vemurafenib |
url |
http://link.springer.com/article/10.1186/s12885-017-3721-7 |
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