In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines
We previously used microarrays to show that high expression of <i>DHRS3</i>, <i>NROB1</i>, and <i>CYP26A1</i> predicts favorable NB outcomes. Here, we investigated whether expression of these genes was associated with suppression of NB cell (SK-N-SH, NB12, and TGW...
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MDPI AG
2022-10-01
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| author | Yoko Hiyama Emi Yamaoka Takahiro Fukazawa Masato Kojima Yusuke Sotomaru Eiso Hiyama |
| author_facet | Yoko Hiyama Emi Yamaoka Takahiro Fukazawa Masato Kojima Yusuke Sotomaru Eiso Hiyama |
| author_sort | Yoko Hiyama |
| collection | DOAJ |
| description | We previously used microarrays to show that high expression of <i>DHRS3</i>, <i>NROB1</i>, and <i>CYP26A1</i> predicts favorable NB outcomes. Here, we investigated whether expression of these genes was associated with suppression of NB cell (SK-N-SH, NB12, and TGW) growth. We assessed morphology and performed growth, colony-formation, and migration assays, as well as RNA sequencing. The effects of the transient expression of these genes were also assessed with a tetracycline-controlled expression (Tet-On) system. Gene overexpression reduced cell growth and induced morphological senescence. Gene-expression analysis identified pathways involving cellular senescence and cell adhesion. In these cells, transduced gene dropout occurred during passage, making long-term stable gene transfer difficult. Tet-On-induced gene expression caused more pronounced cell-morphology changes. Specifically, <i>DHRS3</i> and <i>NROB1</i> led to rapid inhibition and arrest of cell growth, though <i>CYP26A1</i> did not affect cell-growth rate or cell cycle. <i>DHRS3</i> arrested the cell cycle by interacting with the all-trans-retinol pathway and drove differentiation and senescence in tumors. Overexpression of these genes reduced the malignant grade of these cells. A new therapeutic strategy might be the induction of these genes, as they suppress the growth of high-risk neuroblastoma and lead to differentiation and senescence. |
| first_indexed | 2024-03-09T21:52:42Z |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T21:52:42Z |
| publishDate | 2022-10-01 |
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| series | Cells |
| spelling | doaj.art-4e7072f86bdc473aab2ca19c380018752023-11-23T20:04:03ZengMDPI AGCells2073-44092022-10-011119317110.3390/cells11193171In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell LinesYoko Hiyama0Emi Yamaoka1Takahiro Fukazawa2Masato Kojima3Yusuke Sotomaru4Eiso Hiyama5Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanBiomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanBiomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanBiomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanBiomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanBiomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, JapanWe previously used microarrays to show that high expression of <i>DHRS3</i>, <i>NROB1</i>, and <i>CYP26A1</i> predicts favorable NB outcomes. Here, we investigated whether expression of these genes was associated with suppression of NB cell (SK-N-SH, NB12, and TGW) growth. We assessed morphology and performed growth, colony-formation, and migration assays, as well as RNA sequencing. The effects of the transient expression of these genes were also assessed with a tetracycline-controlled expression (Tet-On) system. Gene overexpression reduced cell growth and induced morphological senescence. Gene-expression analysis identified pathways involving cellular senescence and cell adhesion. In these cells, transduced gene dropout occurred during passage, making long-term stable gene transfer difficult. Tet-On-induced gene expression caused more pronounced cell-morphology changes. Specifically, <i>DHRS3</i> and <i>NROB1</i> led to rapid inhibition and arrest of cell growth, though <i>CYP26A1</i> did not affect cell-growth rate or cell cycle. <i>DHRS3</i> arrested the cell cycle by interacting with the all-trans-retinol pathway and drove differentiation and senescence in tumors. Overexpression of these genes reduced the malignant grade of these cells. A new therapeutic strategy might be the induction of these genes, as they suppress the growth of high-risk neuroblastoma and lead to differentiation and senescence.https://www.mdpi.com/2073-4409/11/19/3171neuroblastomacell line<i>DHRS3</i><i>NROB1</i><i>CYP26A1</i>transfection |
| spellingShingle | Yoko Hiyama Emi Yamaoka Takahiro Fukazawa Masato Kojima Yusuke Sotomaru Eiso Hiyama In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines Cells neuroblastoma cell line <i>DHRS3</i> <i>NROB1</i> <i>CYP26A1</i> transfection |
| title | In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines |
| title_full | In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines |
| title_fullStr | In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines |
| title_full_unstemmed | In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines |
| title_short | In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines |
| title_sort | in vitro transfection of up regulated genes identified in favorable outcome neuroblastoma into cell lines |
| topic | neuroblastoma cell line <i>DHRS3</i> <i>NROB1</i> <i>CYP26A1</i> transfection |
| url | https://www.mdpi.com/2073-4409/11/19/3171 |
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