Although the administration of retinoids represents an important part of treatment for children suffering from high-risk neuroblastomas, approximately 50% of these patients do not respond to this therapy or develop resistance to retinoids during treatment. Our study focused on the comparative analysis of the expression of five genes and corresponding proteins (DDX39A, HMGA1, HMGA2, HOXC9 and PBX1) that have recently been discussed as possible predictive biomarkers of clinical response to retinoid differentiation therapy. Expression of these five candidate biomarkers was evaluated at both the mRNA and protein level in the same subset of 8 neuroblastoma cell lines after treatment with natural or synthetic retinoids. We found that the cell lines that were HMGA2-positive and/or HOXC9-negative have a reduced sensitivity to retinoids. Furthermore, the experiments revealed that the retinoid-sensitive cell lines showed a uniform pattern of change after treatment with both natural and sensitive retinoids: increased DDX39A and decreased PBX1 protein levels. Our results showed that in NBL cells, these putative protein biomarkers are associated with sensitivity or resistance to retinoids, and their endogenous or induced expression can distinguish between these two phenotypes.

Department of Pathology University Hospital Brno and Faculty of Medicine Masaryk University Jihlavska Czech Republic

Department of Pediatric Oncology University Hospital Brno and Faculty of Medicine Masaryk University Cernopolni Czech Republic

International Clinical Research Center St Anne's University Hospital Pekarska Czech Republic

Laboratory of Tumor Biology Department of Experimental Biology Faculty of Science Masaryk University Kotlarska Czech Republic

Zobrazit více v PubMed

Cheung N-KV, Dyer MA. Neuroblastoma: developmental biology, cancer genomics and immunotherapy. Nat Rev Cancer. 2013. June;13(6):397–411. 10.1038/nrc3526 PubMed DOI PMC

Irwin MS, Park JR. Neuroblastoma: Paradigm for Precision Medicine. Pediatr Clin North Am. 2015. February 1;62(1):225–56. 10.1016/j.pcl.2014.09.015 PubMed DOI

Moreno L, Marshall LV, Pearson ADJ. At the frontier of progress for paediatric oncology: the neuroblastoma paradigm. Br Med Bull. 2013;108:173–88. 10.1093/bmb/ldt033 PubMed DOI

Park JR, Bagatell R, London WB, Maris JM, Cohn SL, Mattay KK, et al. Children’s Oncology Group’s 2013 blueprint for research: neuroblastoma. Pediatr Blood Cancer. 2013. June;60(6):985–93. 10.1002/pbc.24433 PubMed DOI

Masetti R, Biagi C, Zama D, Vendemini F, Martoni A, Morello W, et al. Retinoids in Pediatric Onco-Hematology: the Model of Acute Promyelocytic Leukemia and Neuroblastoma. Adv Ther. 2012. September 1;29(9):747–62. 10.1007/s12325-012-0047-3 PubMed DOI

Dobrotkova V, Chlapek P, Mazanek P, Sterba J, Veselska R. Traffic lights for retinoids in oncology: molecular markers of retinoid resistance and sensitivity and their use in the management of cancer differentiation therapy. BMC Cancer. 2018. November 1;18(1):1059 10.1186/s12885-018-4966-5 PubMed DOI PMC

Hu J, Liu Y-F, Wu C-F, Xu F, Shen Z-X, Zhu Y-M, et al. Long-term efficacy and safety of all-trans retinoic acid/arsenic trioxide-based therapy in newly diagnosed acute promyelocytic leukemia. Proc Natl Acad Sci USA. 2009. March 3;106(9):3342–7. 10.1073/pnas.0813280106 PubMed DOI PMC

Sterba J. Contemporary therapeutic options for children with high risk neuroblastoma. Neoplasma. 2002;49(3):133–40. PubMed

Chlapek P, Slavikova V, Mazanek P, Sterba J, Veselska R. Why Differentiation Therapy Sometimes Fails: Molecular Mechanisms of Resistance to Retinoids. Int J Mol Sci. 2018. January 3;19(1). 10.3390/ijms19010132 PubMed DOI PMC

Duong V, Rochette-Egly C. The molecular physiology of nuclear retinoic acid receptors. From health to disease. Biochim Biophys Acta. 2011. August;1812(8):1023–31. 10.1016/j.bbadis.2010.10.007 PubMed DOI

Freemantle SJ, Spinella MJ, Dmitrovsky E. Retinoids in cancer therapy and chemoprevention: promise meets resistance. Oncogene. 2003. October 20;22(47):7305–15. 10.1038/sj.onc.1206936 PubMed DOI

Otake K, Uchida K, Ide S, Kobayashi Y, Kobayashi I, Kusunoki M. Identification of DDX39A as a Potential Biomarker for Unfavorable Neuroblastoma Using a Proteomic Approach. Pediatr Blood Cancer. 2016. February;63(2):221–7. 10.1002/pbc.25778 PubMed DOI

Giannini G, Di Marcotullio L, Ristori E, Zani M, Crescenzi M, Scarpa S, et al. HMGI(Y) and HMGI-C genes are expressed in neuroblastoma cell lines and tumors and affect retinoic acid responsiveness. Cancer Res. 1999. May 15;59(10):2484–92. PubMed

Mao L, Ding J, Zha Y, Yang L, McCarthy BA, King W, et al. HOXC9 links cell-cycle exit and neuronal differentiation and is a prognostic marker in neuroblastoma. Cancer Res. 2011. June 15;71(12):4314–24. 10.1158/0008-5472.CAN-11-0051 PubMed DOI PMC

Schnabel CA, Selleri L, Cleary ML. Pbx1 is essential for adrenal development and urogenital differentiation. Genesis. 2003. November;37(3):123–30. 10.1002/gene.10235 PubMed DOI

Phelan ML, Featherstone MS. Distinct HOX N-terminal arm residues are responsible for specificity of DNA recognition by HOX monomers and HOX.PBX heterodimers. J Biol Chem. 1997. March 28;272(13):8635–43. 10.1074/jbc.272.13.8635 PubMed DOI

Shah N, Wang J, Selich-Anderson J, Graham G, Siddiqui H, Li X, et al. PBX1 is a favorable prognostic biomarker as it modulates 13-cis retinoic acid-mediated differentiation in neuroblastoma. Clin Cancer Res. 2014. August 15;20(16):4400–12. 10.1158/1078-0432.CCR-13-1486 PubMed DOI PMC

Veselska R, Kuglik P, Cejpek P, Svachova H, Neradil J, Loja T, et al. Nestin expression in the cell lines derived from glioblastoma multiforme. BMC Cancer. 2006. February 2;6:32 10.1186/1471-2407-6-32 PubMed DOI PMC

Chlapek P, Redova M, Zitterbart K, Hermanova M, Sterba J, Veselska R. Enhancement of ATRA-induced differentiation of neuroblastoma cells with LOX/COX inhibitors: an expression profiling study. J Exp Clin Cancer Res. 2010. May 11;29:45 10.1186/1756-9966-29-45 PubMed DOI PMC

Cerignoli F, Ambrosi C, Mellone M, Assimi I, Di MARCOTULLIO L, Gulino A, et al. HMGA Molecules in Neuroblastic Tumors. Ann N Y Acad Sci. 2004. December 1;1028(1):122–32. 10.1196/annals.1322.013 PubMed DOI

Wang X, Yang L, Choi J-H, Kitamura E, Chang C-S, Ding J, et al. Genome-wide analysis of HOXC9-induced neuronal differentiation of neuroblastoma cells. Genomics Data. 2014. December 1;2:50–2. 10.1016/j.gdata.2014.04.002 PubMed DOI PMC

Xuan F, Huang M, Liu W, Ding H, Yang L, Cui H. Homeobox C9 suppresses Beclin1-mediated autophagy in glioblastoma by directly inhibiting the transcription of death-associated protein kinase 1. Neuro Oncol. 2016;18(6):819–29. 10.1093/neuonc/nov281 PubMed DOI PMC

Hur H, Lee J-Y, Yang S, Kim JM, Park AE, Kim MH. HOXC9 Induces Phenotypic Switching between Proliferation and Invasion in Breast Cancer Cells. J Cancer. 2016;7(7):768–73. 10.7150/jca.13894 PubMed DOI PMC

Kocak H, Ackermann S, Hero B, Kahlert Y, Oberthuer A, Juraeva D, et al. Hox-C9 activates the intrinsic pathway of apoptosis and is associated with spontaneous regression in neuroblastoma. Cell Death Dis. 2013. April 11;4:e586 10.1038/cddis.2013.84 PubMed DOI PMC

Magnani L, Patten DK, Nguyen VTM, Hong S-P, Steel JH, Patel N, et al. The pioneer factor PBX1 is a novel driver of metastatic progression in ERα-positive breast cancer. Oncotarget. 2015. September 8;6(26):21878–91. 10.18632/oncotarget.4243 PubMed DOI PMC

Jung J-G, Shih I-M, Park JT, Gerry E, Kim TH, Ayhan A, et al. Ovarian Cancer Chemoresistance Relies on the Stem Cell Reprogramming Factor PBX1. Cancer Res. 2016. 01;76(21):6351–61. 10.1158/0008-5472.CAN-16-0980 PubMed DOI PMC

Serotonin limits generation of chromaffin cells during adrenal organ development

Comparative Analysis of Putative Prognostic and Predictive Markers in Neuroblastomas: High Expression of PBX1 Is Associated With a Poor Response to Induction Therapy