Homeobox B4 gene expression is upregulated by ghrelin through PI3-kinase signaling pathway in rat’s bone marrow stromal cells

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Objective. Ghrelin, a 28 amino acid peptide, has diverse physiological roles. Phosphatidylino-sitol-bisphosphate 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) are involved in some of the recognized actions of ghrelin. It has been shown that ghrelin upregulates HOXB4 gene expression but the real mechanism of this effect is not clear.

Methods. Rat bone marrow stromal cells (BMSCs) were cultured in DMEM. BMSCs were treated with ghrelin (100 μM) for 48 h. Real-time PCR for HOXB4 was performed from Control (untreated BMSCs), BG (BMSCs treated with 100 µM ghrelin), PD (BMSCs treated with 10 µM PD98059, a potent inhibitor of mitogen-activated protein kinase, and 100 µM ghrelin), LY (BM-SCs treated with 10 µM LY294002, a strong inhibitor of phosphoinositide 3-kinase, and 100 µM ghrelin) and SY (BMSCs treated with 10 µM LY294002 plus 10 µM PD98059, and 100 µM ghrelin) groups. Relative gene expression changes were determined using Relative expression software tool 9 (REST 9).

Results. HOXB4 gene has been overexpressed in ghrelin-treated BMSCs (p<0.05). PI3K inhi-bition by LY294002 significantly downregulated the ghrelin-induced overexpression of HOXB4 (p<0.05).

Conclusion. We can conclude that ghrelin, through PI3K/Akt pathway, may improve BMSC transplantation potency by reducing its apoptosis. Moreover, upregulating HOXB4 in BMSC and its possible differentiation to HSCs might in the future open the doors to new treatment for hematologic disorders. Therefore, activating the PI3K/Akt pathway, instead of using a non-specific inducer, could be the principal point to increase the efficiency of BMSC-based cell therapies in the future.

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  • Abdanipour A Shahsavandi B Dadkhah M Alipour M Feizi H. The antiapoptotic effect of ghrelin in the H2O2 treated Bone Marrow-derived Mesenchymal Stem cells of rat. J Zanjan Univ Med Sci Health Serv 25 58–68 2017.

  • Abdanipour A Shahsavandi B Alipour M Feizi H. Ghrelin upregulates HOXB4 gene expression in the rat BMSCs. Cell J 20 183–187 2018a.

  • Abdanipour A Dadkhah M Alipour M Feizi H. Effect of ghrelin on caspase 3 and Bcl2 gene expression in H2O2 treated rat’s bone marrow stromal cells. Adv Pharm Bull 8 429–435 2018b.

  • Albarran-Zeckler RG Smith RG. The ghrelin receptors (GHS-R1a and GHS-R1b). Endocr Dev 25 5–15 2013.

  • Anthony BA Link DC. Regulation of hematopoietic stem cells by bone marrow stromal cells. Trends Immunol 35 32–37 2014.

  • Antonchuk J Sauvageau G Humphries RK. HOXB4 overexpression mediates very rapid stem cell regeneration and competitive hematopoietic repopulation. Exp Hemato 29 1125–1134 2001.

  • Antonchuk J Sauvageau G Humphries RK. HOXB4-induced expansion of adult hematopoietic stem cells ex vivo. Cell 109 39–45 2002.

  • Baldanzi G Filigheddu N Cutrupi S Catapano F Bonissoni S Fubini A Malan D Baj G Granata R Broglio F Papotti M Surico N Bussolino F Isgaard J Deghenghi R Sinigaglia F Prat M Muccioli G Ghigo E Graziani A. Ghrelin and des-acyl ghrelin inhibit cell death in cardiomyocytes and endothelial cells through ERK1/2 and PI 3-kinase/AKT. J Cell Biol 159 1029–1037 2002.

  • Bi L Wang G Yang D Li S Liang B Han Z. Effects of autologous bone marrow-derived stem cell mobilization on acute tubular necrosis and cell apoptosis in rats. Exp Ther Med 10 851–856 2015.

  • Bianco P Riminucci M Gronthos S Robey PG. Bone marrow stromal stem cells: nature biology and potential applications. Stem Cells 19 180–192 2001.

  • Chung H Seo S Moon M Park S. Phosphatidylinositol-3-kinase/Akt/glycogen synthase kinase-3 beta and ERK1/2 pathways mediate protective effects of acylated and unacylated ghrelin against oxygen-glucose deprivationinduced apoptosis in primary rat cortical neuronal cells. J Endocrinol 198 511–521 2008.

  • Chung H Chung HY Bae CW Kim CJ Park S. Ghrelin suppresses tunicamycinor thapsigargin-triggered endoplasmic reticulum stress-mediated apoptosis in primary cultured rat cortical neuronal cells. Endocr J 58 409–420 2011.

  • Chung H Li E Kim Y Kim S Park S. Multiple signaling pathways mediate ghrelin-induced proliferation of hippocampal neural stem cells. J Endocrinol 218 49–59 2013.

  • Daniels TR Neacato II Rodriguez JA Pandha HS Morgan R Penichet ML. Disruption of HOX activity leads to cell death that canbe enhanced by the interference of iron uptake in malignant B cells. Leukemia 24 1555–1565 2010.

  • Forrester LM Jackson M. Mechanism of action of HOXB4 on the hematopoietic differentiation of embryonic stem cells. Stem Cells 30 379–385 2012.

  • Gan Y Shi C Inge L Hibner M Balducci J Huang Y. Differential roles of ERK and Akt pathways in regulation of EGFR-mediated signaling and motility in prostate cancer cells. Oncogene 29 4947–4958 2010.

  • Han D Huang W Ma S Chen J Gao L Liu T Zhang R Li X Li C Fan M Chen Y Cao F. Ghrelin improves functional survival of engrafted adipose-derived mesenchymal stem cells in ischemic heart through PI3K/Akt signaling pathway. Biomed Res Int 2015 858349 2015.

  • Hu X Yu SP Fraser JL Lu Z Ogle ME Wang JA Wei L. Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis. J Thorac Cardiovasc Surg 135 799–808 2008.

  • Hu C Huang L Gest C Xi X Janin A Soria C Li H Lu H. Opposite regulation by PI3K/Akt and MAPK/ERK pathways of tissue factor expression cell-associated procoagulant activity and invasiveness in MDA-MB-231 cells. J Hematol Oncol 5 16 2012.

  • Jackson M Axton RA Taylor AH Wilson JA Gordon-Keylock SA Kokkaliaris KD Brickman JM Schulz H Hummel O Hubner N Forrester LM. HOXB4 can enhance the differentiation of embryonic stem cells by modulating the hematopoietic niche. Stem Cells 30 150–160 2012.

  • Kanoski SE Fortin SM Ricks KM Grill HJ. Ghrelin signaling in the ventral hippocampus stimulates learned and motivational aspects of feeding via PI3K-Akt signaling. Biol Psychiatry 73 915–923 2013.

  • Kirito K Fox N Kaushansky K. Thrombopoietin stimulates Hoxb4 expression: an explanation for the favorable effects of TPO on hematopoietic stem cells. Blood 102 3172–3178 2003.

  • Kojima M Kangawa K. Ghrelin: structure and function. Physiol Rev 85 495–522 2005.

  • Lee ER Kim JY Kang YJ Ahn JY Kim JH Kim BW Choi HY Jeong MY Cho SG. Interplay between PI3K/Akt and MAPK signaling pathways in DNA-damaging drug-induced apoptosis. Biochim Biophys Acta 1763 958–968 2006.

  • Lee GS Kim BS Sheih JH Moore MA. Forced expression of HoxB4 enhances hematopoietic differentiation by human embryonic stem cells. Mol Cells 25 487–493 2008.

  • Li E Chung H Kim Y Kim DH Ryu JH Sato T Kojima M Park S. Ghrelin directly stimulates adult hippocampal neurogenesis: implications for learning and memory. Endocr J 60 781–789 2013.

  • Mahmood A Lu D Chopp M. Marrow stromal cell transplantation after traumatic brain injury promotes cellular proliferation within the brain. Neurosurgery 55 1185–1193 2004.

  • Moon M Kim S Hwang L Park S. Ghrelin regulates hippocampal neurogenesis in adult mice. Endocrine J 56 525–531 2009.

  • Murata M Okimura Y Iida K Matsumoto M Sowa H Kaji H Kojima M Kangawa K Chihara K. Ghrelin modulates the downstream molecules of insulin signaling in hepatoma cells. J Biol Chem 277 5667–5674 2002.

  • Park SW Won KJ Lee YS Kim HS Kim YK Lee HW Kim B Lee BH Kim JH Kim DK. Increased HoxB4 inhibits apoptotic cell death in Pro-B cells. Korean J Physiol Pharmacol 16 265–271 2012.Potier E Ferreira E Meunier A Sedel L Logeart-Avramoglou D Petite H. Prolonged hypoxia concomitant with serum deprivation induces massive human mesenchymal stem cell death. Tissue Eng 13 1325–1331 2007.

  • Svingen T Tonissen KF. Hox transcription factors and their elusive mammalian gene targets. Heredity 97 88–96 2006.

  • Thorsteinsdottir U Sauvageau G Humphries RK. Enhanced in vivo regenerative potential of HOXB4-transduced hematopoietic stem cells with regulation of their pool size. Blood 94 2605–2612 1999.

  • van der Lely AJ Tschop M Heiman ML Ghigo E. Biological physiological pathophysiological and pharmacological aspects of ghrelin. Endocr Rev 25 426–457 2004.

  • Yang J Liu GQ Wei R Hou WF Gao MJ Zhu MX Wang HN Chen GA Hong TP. Ghrelin promotes differentiation of human embryonic stem cells into cardiomyocytes. Acta Pharmacol Sin 32 1239–1245 2011.

  • Yin Y Li Y Zhang W. The growth hormone secretagogue receptor: its intracellular signaling and regulation. Int J Mol Sci 15 4837–4855 2014.

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