Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth
Abstract The protein alpha-synuclein (α-syn) is unusual because, depending on its conformation and the type of cell in which it is expressed, it is pro-death or pro-survival, triggering neurodegeneration in Parkinson’s disease and enhancing cell survival of some melanomas. To probe the function of α...
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Nature Portfolio
2021-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-84443-y |
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| author | Sahar Shekoohi Santhanasabapathy Rajasekaran Dhaval Patel Shu Yang Wang Liu Shile Huang Xiuping Yu Stephan N. Witt |
| author_facet | Sahar Shekoohi Santhanasabapathy Rajasekaran Dhaval Patel Shu Yang Wang Liu Shile Huang Xiuping Yu Stephan N. Witt |
| author_sort | Sahar Shekoohi |
| collection | DOAJ |
| description | Abstract The protein alpha-synuclein (α-syn) is unusual because, depending on its conformation and the type of cell in which it is expressed, it is pro-death or pro-survival, triggering neurodegeneration in Parkinson’s disease and enhancing cell survival of some melanomas. To probe the function of α-syn in melanoma, we used CRISPR/Cas9 to knockout SNCA, the gene that codes for α-syn, in SK-Mel-28 melanoma cells. The SNCA-knockout clones in culture exhibited a decrease in the transferrin receptor 1 (TfR1), an increase in ferritin, an increase of reactive oxygen species and proliferated slower than control cells. These SNCA-knockout clones grafted into SCID mice grew significantly slower than the SK-Mel-28 control cells that expressed α-syn. In the excised SNCA-knockout xenografts, TfR1 decreased 3.3-fold, ferritin increased 6.2-fold, the divalent metal ion transporter 1 (DMT1) increased threefold, and the iron exporter ferroportin (FPN1) decreased twofold relative to control xenografts. The excised SNCA-KO tumors exhibited significantly more ferric iron and TUNEL staining relative to the control melanoma xenografts. Collectively, depletion of α-syn in SK-Mel-28 cells dysregulates cellular iron metabolism, especially in xenografts, yielding melanoma cells that are deficient in TfR1 and FPN1, that accumulate ferric iron and ferritin, and that undergo apoptosis relative to control cells expressing α-syn. |
| first_indexed | 2024-12-17T20:29:05Z |
| format | Article |
| id | doaj.art-97f081a407604a379445ec7aeb245044 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-17T20:29:05Z |
| publishDate | 2021-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-97f081a407604a379445ec7aeb2450442022-12-21T21:33:38ZengNature PortfolioScientific Reports2045-23222021-03-0111111510.1038/s41598-021-84443-yKnocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growthSahar Shekoohi0Santhanasabapathy Rajasekaran1Dhaval Patel2Shu Yang3Wang Liu4Shile Huang5Xiuping Yu6Stephan N. Witt7Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterDepartment of Biochemistry and Molecular Biology, Louisiana State University Health Sciences CenterAbstract The protein alpha-synuclein (α-syn) is unusual because, depending on its conformation and the type of cell in which it is expressed, it is pro-death or pro-survival, triggering neurodegeneration in Parkinson’s disease and enhancing cell survival of some melanomas. To probe the function of α-syn in melanoma, we used CRISPR/Cas9 to knockout SNCA, the gene that codes for α-syn, in SK-Mel-28 melanoma cells. The SNCA-knockout clones in culture exhibited a decrease in the transferrin receptor 1 (TfR1), an increase in ferritin, an increase of reactive oxygen species and proliferated slower than control cells. These SNCA-knockout clones grafted into SCID mice grew significantly slower than the SK-Mel-28 control cells that expressed α-syn. In the excised SNCA-knockout xenografts, TfR1 decreased 3.3-fold, ferritin increased 6.2-fold, the divalent metal ion transporter 1 (DMT1) increased threefold, and the iron exporter ferroportin (FPN1) decreased twofold relative to control xenografts. The excised SNCA-KO tumors exhibited significantly more ferric iron and TUNEL staining relative to the control melanoma xenografts. Collectively, depletion of α-syn in SK-Mel-28 cells dysregulates cellular iron metabolism, especially in xenografts, yielding melanoma cells that are deficient in TfR1 and FPN1, that accumulate ferric iron and ferritin, and that undergo apoptosis relative to control cells expressing α-syn.https://doi.org/10.1038/s41598-021-84443-y |
| spellingShingle | Sahar Shekoohi Santhanasabapathy Rajasekaran Dhaval Patel Shu Yang Wang Liu Shile Huang Xiuping Yu Stephan N. Witt Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth Scientific Reports |
| title | Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| title_full | Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| title_fullStr | Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| title_full_unstemmed | Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| title_short | Knocking out alpha-synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| title_sort | knocking out alpha synuclein in melanoma cells dysregulates cellular iron metabolism and suppresses tumor growth |
| url | https://doi.org/10.1038/s41598-021-84443-y |
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