Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells
Cutaneous melanoma is often resistant to therapy due to its high plasticity, as well as its ability to metabolise chemotherapeutic drugs. Sphingolipid signalling plays a pivotal role in its progression and metastasis. One of the ways melanoma alters sphingolipid rheostat is via over-expression of ly...
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MDPI AG
2021-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/6/3247 |
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| author | Michele Lai Veronica La Rocca Rachele Amato Giulia Freer Mario Costa Pietro Giorgio Spezia Paola Quaranta Giuseppe Lombardo Daniele Piomelli Mauro Pistello |
| author_facet | Michele Lai Veronica La Rocca Rachele Amato Giulia Freer Mario Costa Pietro Giorgio Spezia Paola Quaranta Giuseppe Lombardo Daniele Piomelli Mauro Pistello |
| author_sort | Michele Lai |
| collection | DOAJ |
| description | Cutaneous melanoma is often resistant to therapy due to its high plasticity, as well as its ability to metabolise chemotherapeutic drugs. Sphingolipid signalling plays a pivotal role in its progression and metastasis. One of the ways melanoma alters sphingolipid rheostat is via over-expression of lysosomal acid ceramidase (AC), which catalyses the hydrolysis of pro-apoptotic long-chain ceramides into sphingosine and fatty acid. In this report, we examine the role of acid ceramidase in maintaining cellular homeostasis through the regulation of autophagy and mitochondrial activity in melanoma cell lines. We show that under baseline conditions, wild-type melanoma cells had 3-fold higher levels of the autophagy marker, microtubule-associated proteins 1A/1B light chain 3B (LC3 II), compared to AC-null cells. This difference was further magnified after cell starvation. Moreover, we noticed autophagy impairment in A375 AC-null cells, possibly due to local accumulation of non-metabolized ceramides. Nonetheless, we observed that AC-null cells exhibited a significant increase in mitochondrial membrane potential compared to control cells. Consistent with this observation, we found that, after total starvation, ~30% of AC-null cells undergo apoptosis compared to ~6% of wild-type cells. As expected, AC transfection restored viability in A375 AC-null cells. Together, these findings suggest that AC-null melanoma cells change and adapt their metabolism to survive in the absence of AC, although in a way that does not allow them to cope with the stress of nutrient deprivation. |
| first_indexed | 2024-03-10T13:00:12Z |
| format | Article |
| id | doaj.art-a19448011a15494aa907a5002aceabd6 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T13:00:12Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a19448011a15494aa907a5002aceabd62023-11-21T11:36:34ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-03-01226324710.3390/ijms22063247Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma CellsMichele Lai0Veronica La Rocca1Rachele Amato2Giulia Freer3Mario Costa4Pietro Giorgio Spezia5Paola Quaranta6Giuseppe Lombardo7Daniele Piomelli8Mauro Pistello9Retrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyRetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalySant’Anna School of Advanced Studies, 56100 Pisa, ItalyRetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyInstitute of Neuroscience, Italian National Research Council (CNR), 56100 Pisa, ItalyRetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyRetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyRetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyAnatomy and Neurobiology, University of California, Irvine, California, 829 Health Sciences Rd, Irvine, CA 92617, USARetrovirus Center and Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, 56100 Pisa, ItalyCutaneous melanoma is often resistant to therapy due to its high plasticity, as well as its ability to metabolise chemotherapeutic drugs. Sphingolipid signalling plays a pivotal role in its progression and metastasis. One of the ways melanoma alters sphingolipid rheostat is via over-expression of lysosomal acid ceramidase (AC), which catalyses the hydrolysis of pro-apoptotic long-chain ceramides into sphingosine and fatty acid. In this report, we examine the role of acid ceramidase in maintaining cellular homeostasis through the regulation of autophagy and mitochondrial activity in melanoma cell lines. We show that under baseline conditions, wild-type melanoma cells had 3-fold higher levels of the autophagy marker, microtubule-associated proteins 1A/1B light chain 3B (LC3 II), compared to AC-null cells. This difference was further magnified after cell starvation. Moreover, we noticed autophagy impairment in A375 AC-null cells, possibly due to local accumulation of non-metabolized ceramides. Nonetheless, we observed that AC-null cells exhibited a significant increase in mitochondrial membrane potential compared to control cells. Consistent with this observation, we found that, after total starvation, ~30% of AC-null cells undergo apoptosis compared to ~6% of wild-type cells. As expected, AC transfection restored viability in A375 AC-null cells. Together, these findings suggest that AC-null melanoma cells change and adapt their metabolism to survive in the absence of AC, although in a way that does not allow them to cope with the stress of nutrient deprivation.https://www.mdpi.com/1422-0067/22/6/3247acid ceramidaseadjuvant therapymelanomaautophagycancer biologyceramides |
| spellingShingle | Michele Lai Veronica La Rocca Rachele Amato Giulia Freer Mario Costa Pietro Giorgio Spezia Paola Quaranta Giuseppe Lombardo Daniele Piomelli Mauro Pistello Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells International Journal of Molecular Sciences acid ceramidase adjuvant therapy melanoma autophagy cancer biology ceramides |
| title | Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells |
| title_full | Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells |
| title_fullStr | Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells |
| title_full_unstemmed | Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells |
| title_short | Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells |
| title_sort | ablation of acid ceramidase impairs autophagy and mitochondria activity in melanoma cells |
| topic | acid ceramidase adjuvant therapy melanoma autophagy cancer biology ceramides |
| url | https://www.mdpi.com/1422-0067/22/6/3247 |
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