Chordoma and its embryonic determinants
DOI:
https://doi.org/10.26481/marble.2014.v2.308Abstract
Background. Chordomas are rare malignant tumors occurring along the axial skeleton that originate from notochordal remnants. It is hypothesized that embryonic factors are involved in their aetiology. In the present study, four such factors are of interest: EVX1, which mediates posterior patterning of the embryo and is found in prostate. Wnt3a, localized to the placenta, is involved in different proliferation signaling pathways. Twist is a mediator of the epithelial-mesenchymal transition (EMT) and is expressed in the neuroblastoma cell line SHSY 5Y. Finally, brachyury is a crucial EMT mediator and involved in cell cycle regulation. Research question. The research question is tripartite. (I) It is the aim to obtain positive control tissue for validation of embryonic primers for the EVX1, Wnt3a and Twist genes. (II) Second, expression of brachyury mRNA in cultured chordoma cells will be assessed in order to validate the cell line. (III) Last, it will be investigated whether fibroblast conditioned cell culturing medium exerts a positive effect on chordoma proliferation in vitro. Materials and methods. (I) Placenta and prostate samples were obtained from patients in the Maastricht University Hospital. mRNA was isolated from prostate, placenta and SHSY 5Y cells and cDNA of the genes of interest was generated after which PCR and gel electrophoresis was employed to yield amplicon bands. (II) Chordoma cells were cultured and brachyury mRNA expression was assessed by PCR assay. (III) Cultured chordoma cells were grown in enriched DMEM or in enriched DMEM containing 33% fibroblast conditioned medium and proliferation was assessed. Results. (I) Both EVX1 and Twist are positive in prostate and SHSY 5Y respectively. Wnt3a is not positive in placenta. (II) Brachyury expression is either strongly downregulated or not expressed in two slowly growing chordoma cell cultures. (III) First data indicate enriched DMEM containing 33% fibroblast conditioned medium does not stimulate chordoma proliferation in vitro as compared to enriched DMEM without fibroblast conditioned medium.References
Chan WCW, Au TYK, Tam V, Cheah KSE, Chan D. Coming together is a beginning: the making of an intervertebral disc. Birth Defects Res C Embryo Today. 2014;102(1):83–100.
Pazzaglia UE, Salisbury JR, Byers PD. Development and involution of the notochord in the human spine. J R Soc Med. 1989;82(7):413–5.
Scheil-Bertram S, Kappler R, von Baer A, Hartwig E, Sarkar M, Serra M, et al. Molecular profiling of chordoma. Int J Oncol. 2014;44(4):1041–55.
McMaster ML, Goldstein AM, Bromley CM, Ishibe N, Parry DM. Chordoma: Incidence and survival patterns in the United States, 1973-1995. Cancer Causes Control. 2001;12:1–11.
Miranda JCF, Gardner PA, Snyderman CH, Devaney KO, Mendenhall WM, Su C, et al. Clival chordomas : A pathological , surgical , and radiotherapeutic review. 2014;(June).
Safari M, Khoshnevisan A. An overview of the role of cancer stem cells in spine tumors with a special focus on chordoma. World J Stem Cells. 2014;6(1):53–64.
Fernando RI, Litzinger M, Trono P, Hamilton DH, Schlom J, Palena C. The T-box transcription factor Brachyury promotes epithelial-mesenchymal transition in human tumor cells. J Clin Invest. 2010;120:533–44.
Fernando RI, Castillo MD, Litzinger M, Hamilton DH, Palena C. IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells. Cancer Res. 2011;71:5296–306.
Faiella A, D’Esposito M, Rambaldi M, Acampora D, Balsofiore S, Stornaiuolo A, et al. Isolation and mapping of EVX1, a human homeobox gene homologous to even-skipped, localized at the 5’ end of HOX1 locus on chromosome 7. Nucleic Acids Res. 1991;19:6541–5.
Yun M-S, Kim S-E, Jeon SH, Lee J-S, Choi K-Y. Both ERK and Wnt/beta-catenin pathways are involved in Wnt3ainduced proliferation. J Cell Sci. 2005;118:313–22.
Yamaguchi TP, Takada S, Yoshikawa Y, Wu N, McMahon AP. T (Brachyury) is a direct target of Wnt3a during paraxial mesoderm specification. Genes Dev. 1999;13:3185–90.
Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C, et al. Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell. 2004;117:927–39.
The Human Protein Atlas. EVX1 RNA Expression [Internet]. [cited 2014 Jun 15]. Available from: http://www. proteinatlas.org/ENSG00000106038/tissue
Saitoh T, Hirai M, Katoh M. Molecular cloning and characterization of WNT3A and WNT14 clustered in human chromosome 1q42 region. Biochem Biophys Res Commun. 2001;284(5):1168–75.
The Human Protein Atlas. TWIST1 RNA Expression [Internet]. [cited 2014 Jun 19]. Available from: http://www. proteinatlas.org/ENSG00000122691/cell
Kang Y, Massagué J. Epithelial-mesenchymal transitions: Twist in development and metastasis. Cell. 2004. p. 277–9.
Pozharskaya V, Torres-González E, Rojas M, Gal A, Amin M, Dollard S, et al. Twist: A regulator of epithelialmesenchymal transition in lung fibrosis. PLoS One. 2009;4.
The Human Protein Atlas. Wnt3a RNA Expression [Internet]. [cited 2014 Jun 15]. Available from: http://www.proteinatlas.org/ENSG00000154342/tissue
Rinner B. Establishment and detailed functional and molecular genetic characterisation of a novel sacral chordoma cell line, MUG-Chor1. International Journal of Oncology. 2011.
Liu X, Nielsen GP, Rosenberg AE, Waterman PR, Yang W, Choy E, et al. Establishment and characterization of a novel chordoma cell line: CH22. J Orthop Res. 2012;30(10):1666–73.
Nelson AC, Pillay N, Henderson S, Presneau N, Tirabosco R, Halai D, et al. An integrated functional genomics approach identifies the regulatory network directed by brachyury (T) in chordoma. J Pathol. 2012;228:274–85.
Hsu W, Mohyeldin A, Shah SR, ap Rhys CM, Johnson LF, Sedora-Roman NI, et al. Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target. J Neurosurg. 2011 Oct;115(4):760–9.
Presneau N, Shalaby A, Ye H, Pillay N, Halai D, Idowu B, et al. Role of the transcription factor T (brachyury) in the pathogenesis of sporadic chordoma: A genetic and functional-based study. J Pathol. 2011;223:327–35.
