PKNOX2 expression and regulation in the bone marrow mesenchymal stem cells of Fanconi anemia patients and healthy donors
- 86 Downloads
HOX and TALE transcription factors are important regulators of development and homeostasis in determining cellular identity. Deregulation of this process may drive cancer progression. The aim of this study was to investigate the expression of these transcription factors in the bone marrow derived mesenchymal stem cells (BM-MSCs) of Fanconi anemia (FA) patients, which is a cancer-predisposing disease. Expression levels of HOX and TALE genes in BM-MSCs were obtained from FA patients and healthy donors by RT-qPCR and highly conserved expression levels were observed between patient and donor cells, except PKNOX2, which is a member of TALE class. PKNOX2 was significantly downregulated in FA cells compared to donors (P < 0.05). PKNOX2 expression levels did not change with diepoxybutane (DEB), a DNA crosslinking agent, in either donor or FA cells except one patient’s with a truncation mutation of FANCA. A difference of PKNOX2 protein level was not obtained between FA patient and donor BM-MSCs by western blot analysis. When human TGF-β1 (rTGF-β1) recombinant protein was provided to the cultures, PKNOX2 as well as TGF-β1 expression increased both in FA and donor BM-MSCs in a dose dependent manner. 5 ng/mL rTGF-β stimulation had more dominant effect on the gene expression of donor BM-MSCs compared to FA cells. Decreased PKNOX2 expression in FA BM-MSCs may provide new insights into the molecular pathophysiology of the disease and TGF-β1 levels of the microenvironment may be the cause of PKNOX2 downregulation.
KeywordsPKNOX2 HOX genes TALE class TGF-β1 Fanconi anemia Bone marrow mesenchymal stem cells
This study was supported by The Scientific and Technological Research Council of Turkey (TUBITAK; Project No: 110S021 in conjunction with EU COST Action BM0805 designated as ‘HOX and TALE transcription factors in Development and Disease’ and TUBITAK Project No: 214Z033). The data in this study is a part of Ilgin Cagnan’s Ph.D. thesis.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 10.Yamamoto M, Takai D, Yamamoto F (2003) Comprehensive expression profiling of highly homologous 39 hox genes in 26 different human adult tissues by the modified systematic multiplex RT-pCR method reveals tissue-specific expression pattern that suggests an important role of chromosomal structure in the regulation of Hox gene expression in adult tissues. Gene Expr 11:199–210CrossRefGoogle Scholar
- 13.Bhattacharjee S, Nandi S (2018) Rare genetic diseases with defects in DNA repair: opportunities and challenges in orphan drug development for targeted cancer therapy. Cancers 10(298). https://doi.org/10.3390/cancers10090298
- 16.Zhan-He W (2013) The concept and practice of Fanconi Anemia: from the clinical bedside to the laboratory bench. Transl Pediatr 2:112–119. https://doi.org/10.3978/j.issn.2224-4336.2013.07.01 Google Scholar
- 18.Auerbach AD (1993) Fanconi anemia diagnosis and the diepoxybutane (DEB) test. Exp Hematol 21:731–733Google Scholar
- 24.Cagnan I (2018) HOX and TALE transcription factors in Fanconi anemia bone-marrow mesenchymal stem cells: gene expression and protein interactions. Dissertation, Hacettepe UniversityGoogle Scholar
- 29.Waljee AK, Mukherjee A, Singal AG et al (2013) Comparison of imputation methods for missing laboratory data in medicine. BMJ Open 3. https://doi.org/10.1136/bmjopen-2013-002847
- 30.Davarinejad H (2017) Quantifications of western blots with Image J. http://www.yorku.ca/yisheng/Internal/Protocols/ImageJ.pdf. (Accessed 01 October 2017)
- 36.Ramos TL, Sánchez-Abarca LI, Rosón-Burgo B et al (2017) Mesenchymal stromal cells (MSC) from JAK2 + myeloproliferative neoplasms differ from normal MSC and contribute to the maintenance of neoplastic hematopoiesis. PLoS ONE 12:e0182470. https://doi.org/10.1371/journal.pone.0182470 CrossRefGoogle Scholar
- 39.Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, Saffrich R, Benes V, Blake J, Pfiser S, Ecstein V et al (2008) Replicative senescence of mesenchymal stem cells: a continuous and organized process. PLoS ONE 3. https://doi.org/10.1371/journal.pone.0002213
- 51.Li J, Tripathi BJ, Chalam KV, Tripathi RC (1996) Transforming growth factor-beta 1 and -beta 2 positively regulate TGF-beta 1 mRNA expression in trabecular cells. Invest Ophthalmol Vis Sci 37:2778–2782Google Scholar