{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T10:59:31Z","timestamp":1789037971681,"version":"build-2803163510"},"reference-count":24,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2022,10,10]],"date-time":"2022-10-10T00:00:00Z","timestamp":1665360000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"AMED","award":["P-DIRECT"],"award-info":[{"award-number":["P-DIRECT"]}]},{"name":"AMED","award":["JP18CK0106332"],"award-info":[{"award-number":["JP18CK0106332"]}]},{"name":"AMED","award":["JP18LK0201066"],"award-info":[{"award-number":["JP18LK0201066"]}]},{"name":"AMED","award":["JP19cm0106502"],"award-info":[{"award-number":["JP19cm0106502"]}]},{"name":"AMED","award":["15H02567"],"award-info":[{"award-number":["15H02567"]}]},{"name":"AMED","award":["16H02778"],"award-info":[{"award-number":["16H02778"]}]},{"name":"MEXT","award":["P-DIRECT"],"award-info":[{"award-number":["P-DIRECT"]}]},{"name":"MEXT","award":["JP18CK0106332"],"award-info":[{"award-number":["JP18CK0106332"]}]},{"name":"MEXT","award":["JP18LK0201066"],"award-info":[{"award-number":["JP18LK0201066"]}]},{"name":"MEXT","award":["JP19cm0106502"],"award-info":[{"award-number":["JP19cm0106502"]}]},{"name":"MEXT","award":["15H02567"],"award-info":[{"award-number":["15H02567"]}]},{"name":"MEXT","award":["16H02778"],"award-info":[{"award-number":["16H02778"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>We previously used microarrays to show that high expression of DHRS3, NROB1, and CYP26A1 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, DHRS3 and NROB1 led to rapid inhibition and arrest of cell growth, though CYP26A1 did not affect cell-growth rate or cell cycle. DHRS3 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.<\/jats:p>","DOI":"10.3390\/cells11193171","type":"journal-article","created":{"date-parts":[[2022,10,10]],"date-time":"2022-10-10T05:12:21Z","timestamp":1665378741000},"page":"3171","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["In Vitro Transfection of Up-Regulated Genes Identified in Favorable-Outcome Neuroblastoma into Cell Lines"],"prefix":"10.3390","volume":"11","author":[{"given":"Yoko","family":"Hiyama","sequence":"first","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emi","family":"Yamaoka","sequence":"additional","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Fukazawa","sequence":"additional","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masato","family":"Kojima","sequence":"additional","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"},{"name":"Department of Pediatric Surgery, Hiroshima University Hospital, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusuke","family":"Sotomaru","sequence":"additional","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9179-5037","authenticated-orcid":false,"given":"Eiso","family":"Hiyama","sequence":"additional","affiliation":[{"name":"Biomedical Science Division, Natural Science Center for Basic Research and Development (N-BARD), Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"},{"name":"Department of Pediatric Surgery, Hiroshima University Hospital, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8551, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/s00383-003-1077-3","article-title":"Expression profiling of favorable and unfavorable neuroblastomas","volume":"20","author":"Hiyama","year":"2004","journal-title":"Pediatric Surg. Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1042\/BJ20081834","article-title":"High incidence of DNA mutations and gene amplifications of the ALK gene in advanced sporadic neuroblastoma tumours","volume":"416","author":"Caren","year":"2008","journal-title":"Biochem. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1007\/s00383-009-2448-1","article-title":"Evaluation of genes identified by microarray analysis in favorable neuroblastoma","volume":"25","author":"Kamei","year":"2009","journal-title":"Pediatric Surg. Int."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"31477","DOI":"10.1074\/jbc.M113.514984","article-title":"Dhrs3 protein attenuates retinoic acid signaling and is required for early embryonic patterning","volume":"288","author":"Kam","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"C1146","DOI":"10.1152\/ajpcell.00034.2009","article-title":"Retinoic acid-induced nNOS expression depends on a novel PI3K\/Akt\/DAX1 pathway in human TGW-nu-I neuroblastoma cells","volume":"297","author":"Nagl","year":"2009","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1242\/dev.00357","article-title":"Developing with lethal RA levels: Genetic ablation of Rarg can restore the viability of mice lacking Cyp26a1","volume":"130","author":"Dolle","year":"2003","journal-title":"Development"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_8","first-page":"156","article-title":"Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol -chloroform extraction","volume":"162","author":"Chromczynski","year":"1987","journal-title":"Anal. Biochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1471","DOI":"10.1038\/sj.bjc.6605014","article-title":"International consensus for neuroblastoma molecular diagnostics: Report from the International Neuroblastoma Risk Group (INRG) Biology Committee","volume":"100","author":"Ambros","year":"2009","journal-title":"Br. J. Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1730","DOI":"10.1016\/j.jpedsurg.2003.08.042","article-title":"Differential gene expression profiles between neuroblastomas with high telomerase activity and low telomerase activity","volume":"38","author":"Hiyama","year":"2003","journal-title":"J. Pediatr. Surg."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1002\/1096-911X(20010101)36:1<67::AID-MPO1017>3.0.CO;2-S","article-title":"Biological characteristics of neuroblastoma with partial deletion in the short arm of chromosome 1","volume":"36","author":"Hiyama","year":"2001","journal-title":"Med. Pediatr. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1627","DOI":"10.1038\/sj.bjc.6605029","article-title":"Consensus criteria for sensitive detection of minimal neuroblastoma cells in bone marrow, blood and stem cell preparations by immunocytology and QRT-PCR: Recommendations by the International Neuroblastoma Risk Group Task Force","volume":"100","author":"Beiske","year":"2009","journal-title":"Br. J. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1038\/nm0395-249","article-title":"Correlating telomerase activity levels with human neuroblastoma outcomes","volume":"1","author":"Hiyama","year":"1995","journal-title":"Nat. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2080","DOI":"10.1016\/j.jpedsurg.2016.09.042","article-title":"Telomere biology including TERT rearrangements in neuroblastoma: A useful indicator for surgical treatments","volume":"51","author":"Kawashima","year":"2016","journal-title":"J. Pediatr. Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1038\/ng.3438","article-title":"TERT rearrangements are frequent in neuroblastoma and identify aggressive tumors","volume":"47","author":"Valentijn","year":"2015","journal-title":"Nat. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1126\/science.aat6768","article-title":"A mechanistic classification of clinical phenotypes in neuroblastoma","volume":"362","author":"Ackermann","year":"2018","journal-title":"Science"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"28343","DOI":"10.1074\/jbc.M111.254227","article-title":"p53-Inducible DHRS3 is an endoplasmic reticulum protein associated with lipid droplet accumulation","volume":"286","author":"Deisenroth","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"21790","DOI":"10.1074\/jbc.273.34.21790","article-title":"Molecular characterization of a novel short-chain dehydrogenase\/reductase that reduces all-trans-retinal","volume":"273","author":"Haeseleer","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1146\/annurev-pathol-011110-130303","article-title":"Retinoids, retinoic acid receptors, and cancer","volume":"6","author":"Tang","year":"2011","journal-title":"Annu. Rev. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4735","DOI":"10.1158\/1078-0432.CCR-07-4372","article-title":"Gene expression profiling of papillary thyroid carcinoma identifies transcripts correlated with BRAF mutational status and lymph node metastasis","volume":"14","author":"Oler","year":"2008","journal-title":"Clin. Cancer Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1080\/1061186X.2019.1710157","article-title":"Liposomal delivery of hydrophobic RAMBAs provides good bioavailability and significant enhancement of retinoic acid signalling in neuroblastoma tumour cells","volume":"28","author":"Bilip","year":"2020","journal-title":"J. Drug Target"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s13073-017-0407-3","article-title":"Retinoic acid and TGF-beta signalling cooperate to overcome MYCN-induced retinoid resistance","volume":"9","author":"Duffy","year":"2017","journal-title":"Genome Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1038\/sj.bjc.6602700","article-title":"The retinoid anticancer signal: Mechanisms of target gene regulation","volume":"93","author":"Liu","year":"2005","journal-title":"Br. J. 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