PMID- 25888913 OWN - NLM STAT- MEDLINE DCOM- 20160209 LR - 20181202 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 6 IP - 1 DP - 2015 Feb 20 TI - Metastatic neuroblastoma cancer stem cells exhibit flexible plasticity and adaptive stemness signaling. PG - 2 LID - 10.1186/s13287-015-0002-8 [doi] LID - 2 AB - INTRODUCTION: High-risk neuroblastoma (HR-NB) presenting with hematogenous metastasis is one of the most difficult cancers to cure. Patient survival is poor. Aggressive tumors contain populations of rapidly proliferating clonogens that exhibit stem cell properties, cancer stem cells (CSCs). Conceptually, CSCs that evade intensive multimodal therapy dictate tumor progression, relapse/recurrence, and poor clinical outcomes. Herein, we investigated the plasticity and stem-cell related molecular response of aggressive metastatic neuroblastoma cells that fit the CSC model. METHODS: Well-characterized clones of metastatic site-derived aggressive cells (MSDACs) from a manifold of metastatic tumors of clinically translatable HR-NB were characterized for their CSC fit by examining epithelial-to-mesenchymal transition (EMT) (E-cadherin, N-Cadherin), survival (NFkappaB P65, p50, IkappaB and pIkappaB) and drug resistance (ABCG2) by immunoblotting; pluripotency maintenance (Nanog, SOX2) by immunofluorescence; and EMT and stemness related transcription of 93 genes by QPCR profiling. Plasticity of MSDACs under sequential alternation of culture conditions with serum and serum-free stem-cell conditions was assessed by clonal expansion (BrdU incorporation), tumorosphere formation (anchorage independent growth), EMT and stemness related transcriptome (QPCR profiling) and validated with MYC, SOX2, EGFR, NOTCH1 and CXCL2 immunoblotting. RESULTS: HR-NB MSDACs maintained in alternated culture conditions, serum-free stem cell medium to growth medium with serum and vice versa identified its flexible revocable plasticity characteristics. We observed signatures of stem cell-related molecular responses consistent with phenotypic conversions. Successive reintroduction to the favorable niche not only regained identical EMT, self-renewal capacity, pluripotency maintenance, and other stem cell-related signaling events, but also instigated additional events depicting aggressive adaptive plasticity. CONCLUSIONS: Together, these results demonstrated the flexible plasticity of HR-NB MSDACs that typically fit the CSC model, and further identified the intrinsic adaptiveness of the successive phenotype switching that clarifies the heterogeneity of HR-NB. Moreover, the continuous ongoing acquisition of stem cell-related molecular rearrangements may hold the key to the switch from favorable disease to HR-NB. FAU - Pandian, Vijayabaskar AU - Pandian V AD - Department of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. vpandian@ouhsc.edu. FAU - Ramraj, Satishkumar AU - Ramraj S AD - Department of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. sramraj@ouhsc.edu. FAU - Khan, Faizan H AU - Khan FH AD - Department of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. fkhan4@ouhsc.edu. FAU - Azim, Tasfia AU - Azim T AD - Department of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. Tasfia.Azim@OSSM.EDU. FAU - Aravindan, Natarajan AU - Aravindan N AD - Department of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. naravind@ouhsc.edu. LA - eng GR - P20 GM103639/GM/NIGMS NIH HHS/United States GR - 1P20GM103639-01/GM/NIGMS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20150220 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 RN - 0 (ABCG2 protein, human) RN - 0 (ATP Binding Cassette Transporter, Subfamily G, Member 2) RN - 0 (ATP-Binding Cassette Transporters) RN - 0 (Cadherins) RN - 0 (NF-kappa B p50 Subunit) RN - 0 (Neoplasm Proteins) RN - 0 (RELA protein, human) RN - 0 (Transcription Factor RelA) RN - EC 2.7.11.10 (I-kappa B Kinase) SB - IM EIN - Stem Cell Res Ther. 2016;7:32. PMID: 26892617 MH - ATP Binding Cassette Transporter, Subfamily G, Member 2 MH - ATP-Binding Cassette Transporters/metabolism MH - Cadherins/metabolism MH - Cell Plasticity/*physiology MH - Cell Proliferation MH - Epithelial-Mesenchymal Transition/*physiology MH - Gene Expression Regulation, Neoplastic MH - Humans MH - I-kappa B Kinase/metabolism MH - NF-kappa B p50 Subunit/metabolism MH - Neoplasm Metastasis/genetics/pathology MH - Neoplasm Proteins/metabolism MH - Neoplasm Recurrence, Local MH - Neoplastic Stem Cells/*pathology MH - Neuroblastoma/*pathology MH - Signal Transduction MH - Spheroids, Cellular MH - Transcription Factor RelA/metabolism MH - Tumor Cells, Cultured PMC - PMC4396071 EDAT- 2015/04/19 06:00 MHDA- 2016/02/10 06:00 PMCR- 2015/02/20 CRDT- 2015/04/19 06:00 PHST- 2014/10/24 00:00 [received] PHST- 2015/02/03 00:00 [accepted] PHST- 2015/02/03 00:00 [revised] PHST- 2015/04/19 06:00 [entrez] PHST- 2015/04/19 06:00 [pubmed] PHST- 2016/02/10 06:00 [medline] PHST- 2015/02/20 00:00 [pmc-release] AID - 10.1186/s13287-015-0002-8 [pii] AID - 2 [pii] AID - 10.1186/s13287-015-0002-8 [doi] PST - epublish SO - Stem Cell Res Ther. 2015 Feb 20;6(1):2. doi: 10.1186/s13287-015-0002-8.