Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets
Expression of 328 ion channel genes was investigated, by in silico analysis, in 170 human melanoma samples and controls. Ninety-one members of this gene-family (i.e., about 28%) show a significant (<i>p</i> < 0.05) differential expression in melanoma- vs. nevi-biopsies, taken from...
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| Format: | Article |
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MDPI AG
2019-03-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/11/4/446 |
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| author | Daniela D’Arcangelo Francesca Scatozza Claudia Giampietri Paolo Marchetti Francesco Facchiano Antonio Facchiano |
| author_facet | Daniela D’Arcangelo Francesca Scatozza Claudia Giampietri Paolo Marchetti Francesco Facchiano Antonio Facchiano |
| author_sort | Daniela D’Arcangelo |
| collection | DOAJ |
| description | Expression of 328 ion channel genes was investigated, by in silico analysis, in 170 human melanoma samples and controls. Ninety-one members of this gene-family (i.e., about 28%) show a significant (<i>p</i> < 0.05) differential expression in melanoma- vs. nevi-biopsies, taken from the GEO database. ROC (receiver operating characteristic) analysis selected 20 genes as potential markers showing the highest discrimination ability of melanoma vs. nevi (AUC > 0.90 and <i>p</i> < 0.0001). These 20 genes underwent a first in silico-validation round in an independent patients-dataset from GEO. A second-in silico-validation step was then carried out on a third human dataset in Oncomine. Finally, five genes were validated, showing extremely high sensitivity and specificity in melanoma detection (>90% in most cases). Such five genes (namely, <i>SCNN1A</i>, <i>GJB3</i>, <i>KCNK7</i>, <i>GJB1</i>, <i>KCNN2</i>) are novel potential melanoma markers or molecular targets, never previously related to melanoma. The “druggable genome„ analysis was then carried out. Miconazole, an antifungal drug commonly used in clinics, is known to target <i>KCNN2</i>, the best candidate among the five identified genes. Miconazole was then tested in vitro in proliferation assays; it dose-dependently inhibited proliferation up to 90% and potently induced cell-death in A-375 and SKMEL-28 melanoma cells, while it showed no effect in control cells. Moreover, specific silencing of <i>KCNN2</i> ion channel was achieved by siRNA transfection; under such condition miconazole strongly increases its anti-proliferative effect. In conclusion, the present study identified five ion channels that can potentially serve as sensitive and specific markers in human melanoma specimens and demonstrates that the antifungal drug miconazole, known to target one of the five identified ion channels, exerts strong and specific anti-melanoma effects in vitro. |
| first_indexed | 2024-03-12T06:56:06Z |
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| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T06:56:06Z |
| publishDate | 2019-03-01 |
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| spelling | doaj.art-7018037f8037463a9588c3d2cb1baa7b2023-09-03T00:01:55ZengMDPI AGCancers2072-66942019-03-0111444610.3390/cancers11040446cancers11040446Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular TargetsDaniela D’Arcangelo0Francesca Scatozza1Claudia Giampietri2Paolo Marchetti3Francesco Facchiano4Antonio Facchiano5Istituto Dermopatico dell’Immacolata (IDI-IRCCS), 00167 Rome, ItalyIstituto Dermopatico dell’Immacolata (IDI-IRCCS), 00167 Rome, ItalyDepartment of Anatomy, Histology, Forensic Medicine and Orthopedics, Unit of Histology and Medical Embryology, Sapienza University of Rome, 00161 Rome, ItalyMedical Oncology, Sapienza, University of Rome, 00161 Rome, ItalyDepartment of Oncology and Molecular Medicine, Istituto Superiore di Sanità (ISS), 00161 Rome, ItalyIstituto Dermopatico dell’Immacolata (IDI-IRCCS), 00167 Rome, ItalyExpression of 328 ion channel genes was investigated, by in silico analysis, in 170 human melanoma samples and controls. Ninety-one members of this gene-family (i.e., about 28%) show a significant (<i>p</i> < 0.05) differential expression in melanoma- vs. nevi-biopsies, taken from the GEO database. ROC (receiver operating characteristic) analysis selected 20 genes as potential markers showing the highest discrimination ability of melanoma vs. nevi (AUC > 0.90 and <i>p</i> < 0.0001). These 20 genes underwent a first in silico-validation round in an independent patients-dataset from GEO. A second-in silico-validation step was then carried out on a third human dataset in Oncomine. Finally, five genes were validated, showing extremely high sensitivity and specificity in melanoma detection (>90% in most cases). Such five genes (namely, <i>SCNN1A</i>, <i>GJB3</i>, <i>KCNK7</i>, <i>GJB1</i>, <i>KCNN2</i>) are novel potential melanoma markers or molecular targets, never previously related to melanoma. The “druggable genome„ analysis was then carried out. Miconazole, an antifungal drug commonly used in clinics, is known to target <i>KCNN2</i>, the best candidate among the five identified genes. Miconazole was then tested in vitro in proliferation assays; it dose-dependently inhibited proliferation up to 90% and potently induced cell-death in A-375 and SKMEL-28 melanoma cells, while it showed no effect in control cells. Moreover, specific silencing of <i>KCNN2</i> ion channel was achieved by siRNA transfection; under such condition miconazole strongly increases its anti-proliferative effect. In conclusion, the present study identified five ion channels that can potentially serve as sensitive and specific markers in human melanoma specimens and demonstrates that the antifungal drug miconazole, known to target one of the five identified ion channels, exerts strong and specific anti-melanoma effects in vitro.https://www.mdpi.com/2072-6694/11/4/446<i>KCNN2</i>ion channelsmelanomamiconazole |
| spellingShingle | Daniela D’Arcangelo Francesca Scatozza Claudia Giampietri Paolo Marchetti Francesco Facchiano Antonio Facchiano Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets Cancers <i>KCNN2</i> ion channels melanoma miconazole |
| title | Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets |
| title_full | Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets |
| title_fullStr | Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets |
| title_full_unstemmed | Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets |
| title_short | Ion Channel Expression in Human Melanoma Samples: In Silico Identification and Experimental Validation of Molecular Targets |
| title_sort | ion channel expression in human melanoma samples in silico identification and experimental validation of molecular targets |
| topic | <i>KCNN2</i> ion channels melanoma miconazole |
| url | https://www.mdpi.com/2072-6694/11/4/446 |
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