Human rhabdomyosarcoma cells express functional erythropoietin receptor: Potential therapeutic implications

Poniewierska-Baran,Agata; Suszynska,Malwina; Sun,Wenyue; Abdelbaset-Ismail,Ahmed; Schneider,Gabriela; Barr,Frederic   G.; Ratajczak,Mariusz  Z.

doi:10.3892/ijo.2015.3184

Human rhabdomyosarcoma cells express functional erythropoietin receptor: Potential therapeutic implications

Authors:
- Agata Poniewierska-Baran
- Malwina Suszynska
- Wenyue Sun
- Ahmed Abdelbaset-Ismail
- Gabriela Schneider
- Frederic G. Barr
- Mariusz Z. Ratajczak
View Affiliations

Affiliations: Stem Cell Institute at James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA, Department of Regenerative Medicine, Warsaw Medical University, Warsaw, Poland, Laboratory of Pathology, National Cancer Institute, Bethesda, MD, USA
Published online on: September 24, 2015 https://doi.org/10.3892/ijo.2015.3184
Pages: 1989-1997

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The erythropoietin receptor (EpoR) is expressed by cells from the erythroid lineage; however, evidence has accumulated that it is also expressed by some solid tumors. This is an important observation, because recombinant erythropoietin (EPO) is employed in cancer patients to treat anemia related to chemo/radiotherapy. In our studies we employed eight rhabdomyosarcoma (RMS) cell lines (three alveolar-type RMS cell lines and five embrional‑type RMS cell lines), and mRNA samples obtained from positive, PAX7-FOXO1‑positive, and fusion-negative RMS patient samples. Expression of EpoR was evaluated by RT-PCR, gene array and FACS. The functionality of EpoR in RMS cell lines was evaluated by chemotaxis, adhesion, and direct cell proliferation assays. In some of the experiments, RMS cells were exposed to vincristine (VCR) in the presence or absence of EPO to test whether EPO may impair the therapeutic effect of VCR. We report for a first time that functional EpoR is expressed in human RMS cell lines as well as by primary tumors from RMS patients. Furthermore, EpoR is detectably expressed in both embryonal and alveolar RMS subtypes. At the functional level, several human RMS cell lines responded to EPO stimulation by enhanced proliferation, chemotaxis, cell adhesion, and phosphorylation of MAPKp42/44 and AKT. Moreover, RMS cells became more resistant to VCR treatment in the presence of EPO. Our findings have important potential clinical implications, indicating that EPO supplementation in RMS patients may have the unwanted side effect of tumor progression.

View Figures

View References

1	Collins MH, Zhao H, Womer RB and Barr FG: Proliferative and apoptotic differences between alveolar rhabdomyosarcoma subtypes: A comparative study of tumors containing PAX3-FKHR or PAX7-FKHR gene fusions. Med Pediatr Oncol. 37:83–89. 2001. View Article : Google Scholar : PubMed/NCBI
2	Sandberg AA, Stone JF, Czarnecki L and Cohen JD: Hematologic masquerade of rhabdomyosarcoma. Am J Hematol. 68:51–57. 2001. View Article : Google Scholar : PubMed/NCBI
3	Davis RJ, D'Cruz CM, Lovell MA, Biegel JA and Barr FG: Fusion of PAX7 to FKHR by the variant t(1;13)(p36;q14) translocation in alveolar rhabdomyosarcoma. Cancer Res. 54:2869–2872. 1994.PubMed/NCBI
4	Schneider G, Bowser MJ, Shin DM, Barr FG and Ratajczak MZ: The paternally imprinted DLK1-GTL2 locus is differentially methylated in embryonal and alveolar rhabdomyosarcomas. Int J Oncol. 44:295–300. 2014.
5	Liang K, Esteva FJ, Albarracin C, Stemke-Hale K, Lu Y, Bianchini G, Yang CY, Li Y, Li X, Chen CT, et al: Recombinant human erythropoietin antagonizes trastuzumab treatment of breast cancer cells via Jak2-mediated Src activation and PTEN inactivation. Cancer Cell. 18:423–435. 2010. View Article : Google Scholar : PubMed/NCBI
6	Todaro M, Turdo A, Bartucci M, Iovino F, Dattilo R, Biffoni M, Stassi G, Federici G, De Maria R and Zeuner A: Erythropoietin activates cell survival pathways in breast cancer stem-like cells to protect them from chemotherapy. Cancer Res. 73:6393–6400. 2013. View Article : Google Scholar : PubMed/NCBI
7	Trost N, Stepisnik T, Berne S, Pucer A, Petan T, Komel R and Debeljak N: Recombinant human erythropoietin alters gene expression and stimulates proliferation of MCF-7 breast cancer cells. Radiol Oncol. 47:382–389. 2013. View Article : Google Scholar : PubMed/NCBI
8	Morais C, Johnson DW, Vesey DA and Gobe GC: Functional significance of erythropoietin in renal cell carcinoma. BMC Cancer. 13:142013. View Article : Google Scholar : PubMed/NCBI
9	Seibold ND, Schild SE, Gebhard MP, Noack F, Schröder U and Rades D: Prognosis of patients with locally advanced squamous cell carcinoma of the head and neck. Impact of tumor cell expression of EPO and EPO-R. Strahlenther Onkol. 189:559–565. 2013. View Article : Google Scholar : PubMed/NCBI
10	Wang L, Li HG, Xia ZS, Wen JM and Lv J: Prognostic significance of erythropoietin and erythropoietin receptor in gastric adenocarcinoma. World J Gastroenterol. 17:3933–3940. 2011. View Article : Google Scholar : PubMed/NCBI
11	Kumar SM, Zhang G, Bastian BC, Arcasoy MO, Karande P, Pushparajan A, Acs G and Xu X: Erythropoietin receptor contributes to melanoma cell survival in vivo. Oncogene. 31:1649–1660. 2012. View Article : Google Scholar :
12	Lopez TV, Lappin TR, Maxwell P, Shi Z, Lopez-Marure R, Aguilar C and Rocha-Zavaleta L: Autocrine/paracrine erythropoietin signalling promotes JAK/STAT-dependent proliferation of human cervical cancer cells. Int J Cancer. 129:2566–2576. 2011. View Article : Google Scholar : PubMed/NCBI
13	Aguilar C, Aguilar C, Lopez-Marure R, Jiménez-Sánchez A and Rocha-Zavaleta L: Co-stimulation with stem cell factor and erythropoietin enhances migration of c-Kit expressing cervical cancer cells through the sustained activation of ERK1/2. Mol Med Rep. 9:1895–1902. 2014.PubMed/NCBI
14	Batra S, Perelman N, Luck LR, Shimada H and Malik P: Pediatric tumor cells express erythropoietin and a functional erythropoietin receptor that promotes angiogenesis and tumor cell survival. Lab Invest. 83:1477–1487. 2003. View Article : Google Scholar : PubMed/NCBI
15	Suszynska M, Poniewierska-Baran A, Gunjal P, Ratajczak J, Marycz K, Kakar SS, Kucia M and Ratajczak MZ: Expression of the erythropoietin receptor by germline-derived cells - further support for a potential developmental link between the germline and hematopoiesis. J Ovarian Res. 7:662014. View Article : Google Scholar : PubMed/NCBI
16	Jacobs JF, Brasseur F, Hulsbergen-van de Kaa CA, van de Rakt MW, Figdor CG, Adema GJ, Hoogerbrugge PM, Coulie PG and de Vries IJ: Cancer-germline gene expression in pediatric solid tumors using quantitative real-time PCR. Int J Cancer. 120:67–74. 2007. View Article : Google Scholar
17	Virchow R: Archiv fuer pathologische anatomie und physiologie und fuer klinische. Medizin. 8:23–54. 1855.(In German).
18	Conheim J: Congenitales, quergestreiftes Muskelsarkon der Nireren. Virchows Arch. 65:64–69. 1875.(In German). View Article : Google Scholar
19	Aloisio GM, Nakada Y, Saatcioglu HD, Peña CG, Baker MD, Tarnawa ED, Mukherjee J, Manjunath H, Bugde A, Sengupta AL, et al: PAX7 expression defines germline stem cells in the adult testis. J Clin Invest. 124:3929–3944. 2014. View Article : Google Scholar : PubMed/NCBI
20	Grymula K, Tarnowski M, Wysoczynski M, Drukala J, Barr FG, Ratajczak J, Kucia M and Ratajczak MZ: Overlapping and distinct role of CXCR7-SDF-1/ITAC and CXCR4-SDF-1 axes in regulating metastatic behavior of human rhabdomyosarcomas. Int J Cancer. 127:2554–2568. 2010. View Article : Google Scholar : PubMed/NCBI
21	Rölfing JH, Baatrup A, Stiehler M, Jensen J, Lysdahl H and Bünger C: The osteogenic effect of erythropoietin on human mesenchymal stromal cells is dose-dependent and involves non-hematopoietic receptors and multiple intracellular signaling pathways. Stem Cell Rev. 10:69–78. 2014. View Article : Google Scholar
22	Broxmeyer HE: Erythropoietin: Multiple targets, actions, and modifying influences for biological and clinical consideration. J Exp Med. 210:205–208. 2013. View Article : Google Scholar : PubMed/NCBI
23	Libura J, Drukala J, Majka M, Tomescu O, Navenot JM, Kucia M, Marquez L, Peiper SC, Barr FG, Janowska-Wieczorek A, et al: CXCR4-SDF-1 signaling is active in rhabdomyosarcoma cells and regulates locomotion, chemotaxis, and adhesion. Blood. 100:2597–2606. 2002. View Article : Google Scholar : PubMed/NCBI
24	Jankowski K, Kucia M, Wysoczynski M, Reca R, Zhao D, Trzyna E, Trent J, Peiper S, Zembala M, Ratajczak J, et al: Both hepatocyte growth factor (HGF) and stromal-derived factor-1 regulate the metastatic behavior of human rhabdomyosarcoma cells, but only HGF enhances their resistance to radiochemotherapy. Cancer Res. 63:7926–7935. 2003.PubMed/NCBI
25	Kang MH, Smith MA, Morton CL, Keshelava N, Houghton PJ and Reynolds CP: National Cancer Institute pediatric preclinical testing program: Model description for in vitro cytotoxicity testing. Pediatr Blood Cancer. 56:239–249. 2011. View Article : Google Scholar
26	Maxwell P, Melendez-Rodríguez F, Matchett KB, Aragones J, Ben-Califa N, Jaekel H, Hengst L, Lindner H, Bernardini A, Brockmeier U, et al: Novel antibodies directed against the human erythropoietin receptor: Creating a basis for clinical implementation. Br J Haematol. 168:429–442. 2015. View Article : Google Scholar
27	Stolze I, Berchner-Pfannschmidt U, Freitag P, Wotzlaw C, Rössler J, Frede S, Acker H and Fandrey J: Hypoxia-inducible erythropoietin gene expression in human neuroblastoma cells. Blood. 100:2623–2628. 2002. View Article : Google Scholar : PubMed/NCBI
28	Ribatti D, Poliani PL, Longo V, Mangieri D, Nico B and Vacca A: Erythropoietin/erythropoietin receptor system is involved in angiogenesis in human neuroblastoma. Histopathology. 50:636–641. 2007. View Article : Google Scholar : PubMed/NCBI
29	Sartelet H, Fabre M, Castaing M, Bosq J, Racu I, Lagonotte E, Scott V, Lecluse Y, Barette S, Michiels S, et al: Expression of erythropoietin and its receptor in neuroblastomas. Cancer. 110:1096–1106. 2007. View Article : Google Scholar : PubMed/NCBI
30	Tarnowski M, Grymula K, Reca R, Jankowski K, Maksym R, Tarnowska J, Przybylski G, Barr FG, Kucia M and Ratajczak MZ: Regulation of expression of stromal-derived factor-1 receptors: CXCR4 and CXCR7 in human rhabdomyosarcomas. Mol Cancer Res. 8:1–14. 2010. View Article : Google Scholar : PubMed/NCBI
31	Tarnowski M, Grymula K, Liu R, Tarnowska J, Drukala J, Ratajczak J, Mitchell RA, Ratajczak MZ and Kucia M: Macrophage migration inhibitory factor is secreted by rhabdomyosarcoma cells, modulates tumor metastasis by binding to CXCR4 and CXCR7 receptors and inhibits recruitment of cancer-associated fibroblasts. Mol Cancer Res. 8:1328–1343. 2010. View Article : Google Scholar : PubMed/NCBI
32	Wysoczynski M, Miekus K, Jankowski K, Wanzeck J, Bertolone S, Janowska-Wieczorek A, Ratajczak J and Ratajczak MZ: Leukemia inhibitory factor: A newly identified metastatic factor in rhabdomyosarcomas. Cancer Res. 67:2131–2140. 2007. View Article : Google Scholar : PubMed/NCBI
33	Schneider G, Bryndza E, Abdel-Latif A, Ratajczak J, Maj M, Tarnowski M, Klyachkin YM, Houghton P, Morris AJ, Vater A, et al: Bioactive lipids S1P and C1P are prometastatic factors in human rhabdomyosarcoma, and their tissue levels increase in response to radio/chemotherapy. Mol Cancer Res. 11:793–807. 2013. View Article : Google Scholar : PubMed/NCBI
34	Schneider G, Sellers ZP, Abdel-Latif A, Morris AJ and Ratajczak MZ: Bioactive lipids, LPC and LPA, are novel prometastatic factors and their tissue levels increase in response to radio/chemotherapy. Mol Cancer Res. 12:1560–1573. 2014. View Article : Google Scholar : PubMed/NCBI