Cytokines secreted by human Wharton's jelly stem cells inhibit the proliferation of ovarian cancer (OVCAR3) cells in vitro

Kalamegam,Gauthaman; Sait,Khalid  Hussein Wali; Anfinan,Nisreen; Kadam,Roaa; Ahmed,Farid; Rasool,Mahmood; Naseer,Mohammad  Imran; Pushparaj,Peter  Natesan; Al‑Qahtani,Mohammed

doi:10.3892/ol.2019.10094

Cytokines secreted by human Wharton's jelly stem cells inhibit the proliferation of ovarian cancer (OVCAR3) cells in vitro

Authors:
- Gauthaman Kalamegam
- Khalid Hussein Wali Sait
- Nisreen Anfinan
- Roaa Kadam
- Farid Ahmed
- Mahmood Rasool
- Mohammad Imran Naseer
- Peter Natesan Pushparaj
- Mohammed Al‑Qahtani
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Affiliations: Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah 21589, Kingdom of Saudi Arabia, Department of Obstetrics and Gynaecology, Faculty of Medicine, King Abdulaziz University Hospital, Jeddah 22252, Kingdom of Saudi Arabia
Published online on: March 1, 2019 https://doi.org/10.3892/ol.2019.10094
Pages: 4521-4531
Copyright: © Kalamegam et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cytokines enhance tumour cell recognition via cytotoxic effector cells and are therefore effectively used in cancer immunotherapy. Mesenchymal stem cells have efficient homing potential and have been used to target and inhibit various types of cancer mediated by the release of soluble/bioactive factors. Initial evaluation of the human Wharton's jelly stem cell conditioned medium (hWJSC‑CM) and cell lysate (hWJSC‑CL) against an ovarian cancer cell line (OVCAR3) demonstrated their inhibitory effect in vitro. The secreted cytokine profile was then studied to understand whether the OVCAR3 inhibitory effect was mediated by the cytokines. Expression of cytokines in OVCAR3 following 48 h treatment with hWJSC extracts, namely the hWJSC‑CM (50%) and hWJSC‑CL (10 µg/ml), was evaluated using multiplex cytokine assay. Paclitaxel (5 nM) was used as a positive control. Cytokines tumour necrosis factor α, interleukin (IL)‑4, IL‑6, IL‑8, IL‑10, IL‑13, IL‑17, IL‑1β and granulocyte colony‑stimulating factor, reported to be involved in tumour growth, invasion and migration, were significantly decreased. Cytokines with antitumour effects, namely IL‑1 receptor antagonist (IL‑1RA), IL‑2, IL‑2 receptor, IL‑5, IL‑7, IL‑12, IL‑15, interferon (IFN)‑α and IFN‑γ, were mildly increased or decreased. Only the increases in IL‑1RA (with paclitaxel, hWJSC‑CM and hWJSC‑CL) and granulocyte‑macrophage colony‑stimulating factor (with hWJSC‑CL) were statistically significant. The chemokines monocyte chemoattractant protein 1, macrophage inflammatory protein (MIP)‑1α, MIP‑1β and Regulated Upon Activation, Normally T‑Expressed, and Secreted were significantly decreased while monokine induced by IFN‑γ, IFN‑γ induced protein 10 and Eotaxin demonstrated mild decreases. The growth factors basic fibroblast growth factor, vascular endothelial growth factor and hepatocyte growth factor were significantly decreased. Heatmaps demonstrated differential fold changes in cytokines and hierarchical cluster analysis revealed 3 major and 7 minor sub‑clusters of associated cytokines, chemokines and growth factors. In conclusion, the hWJSC extracts decreased the expression of oncogenic cytokines, chemokines and growth factors, which mediated the inhibition of OVCAR3 cells in vitro.

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1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar : PubMed/NCBI
2	Yang JY, Yoshihara K, Tanaka K, Hatae M, Masuzaki H, Itamochi H; Cancer Genome Atlas (TCGA) Research Network, ; Takano M, Ushijima K, Tanyi JL, et al: Predicting time to ovarian carcinoma recurrence using protein markers. J Clin Invest. 123:3740–3750. 2013. View Article : Google Scholar : PubMed/NCBI
3	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar : PubMed/NCBI
4	Holschneider CH and Berek JS: Ovarian cancer: Epidemiology, biology, and prognostic factors. Semin Surg Oncol. 19:3–10. 2000. View Article : Google Scholar : PubMed/NCBI
5	Rosenberg L, Palmer JR, Zauber AG, Warshauer ME, Lewis JL Jr, Strom BL, Harlap S and Shapiro S: A case-control study of oral contraceptive use and invasive epithelial ovarian cancer. Am J Epidemiol. 139:654–661. 1994. View Article : Google Scholar : PubMed/NCBI
6	Barbieri F, Bajetto A and Florio T: Role of chemokine network in the development and progression of ovarian cancer: A potential novel pharmacological target. J Oncol. 2010:4269562010. View Article : Google Scholar : PubMed/NCBI
7	Ren L, Xiao L, Hu J, Li Z and Wang Z: MDR1 and MDR3 genes and drug resistance to cisplatin of ovarian cancer cells. J Huazhong Univ Sci Technolog Med Sci. 27:721–724. 2007. View Article : Google Scholar : PubMed/NCBI
8	Wu J, Qu Z, Fei ZW, Wu JH and Jiang CP: Role of stem cell-derived exosomes in cancer. Oncol Lett. 13:2855–2866. 2017. View Article : Google Scholar : PubMed/NCBI
9	Mohr A and Zwacka R: The future of mesenchymal stem cell-based therapeutic approaches for cancer-From cells to ghosts. Cancer Lett. 414:239–249. 2018. View Article : Google Scholar : PubMed/NCBI
10	D'souza N, Burns JS, Grisendi G, Candini O, Veronesi E, Piccinno S, Horwitz EM, Paolucci P, Conte P and Dominici M: MSC and tumors: Homing, differentiation, and secretion influence therapeutic potential. Adv Biochem Eng Biotechnol. 130:209–266. 2012.
11	Sohni A and Verfaillie CM: Mesenchymal stem cells migration homing and tracking. Stem Cells Int. 2013:1307632013. View Article : Google Scholar : PubMed/NCBI
12	Rhee KJ, Lee JI and Eom YW: Mesenchymal stem cell-mediated effects of tumor support or suppression. Int J Mol Sci. 16:30015–30033. 2015. View Article : Google Scholar : PubMed/NCBI
13	Rea IM, Gibson DS, McGilligan V, McNerlan SE, Alexander HD and Ross OA: Age and age-related diseases: Role of inflammation triggers and cytokines. Front Immunol. 9:5862018. View Article : Google Scholar : PubMed/NCBI
14	Jammal MP, Martins-Filho A, Silveira TP, Murta EF and Nomelini RS: Cytokines and prognostic factors in epithelial ovarian cancer. Clin Med Insights Oncol. 10:71–76. 2016. View Article : Google Scholar : PubMed/NCBI
15	Zhang L, Conejo-Garcia JR, Katsaros D, Gimotty PA, Massobrio M, Regnani G, Makrigiannakis A, Gray H, Schlienger K, Liebman MN, et al: Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med. 348:203–213. 2003. View Article : Google Scholar : PubMed/NCBI
16	Fong CY, Subramanian A, Biswas A, Gauthaman K, Srikanth P, Hande MP and Bongso A: Derivation efficiency, cell proliferation, freeze-thaw survival, stem-cell properties and differentiation of human Wharton's jelly stem cells. Reprod Biomed Online. 21:391–401. 2010. View Article : Google Scholar : PubMed/NCBI
17	Gauthaman K, Yee FC, Cheyyatraivendran S, Biswas A, Choolani M and Bongso A: Human umbilical cord Wharton's jelly stem cell (hWJSC) extracts inhibit cancer cell growth in vitro. J Cell Biochem. 113:2027–2039. 2012. View Article : Google Scholar : PubMed/NCBI
18	Gauthaman K, Abbas M, Gari M, Alsehli H, Kadam R, Alkaff M, Chaudhary A, Al-Qahtani M, Abuzenadah A, Kafienah W and Mobasheri A: Pellet culture protects bone marrow derived mesenchymal stem cells from the effects of heat shock. Front Physiol. 7:1802016.PubMed/NCBI
19	Sturn A, Quackenbush J and Trajanoski Z: Genesis: Cluster analysis of microarray data. Bioinformatics. 18:207–208. 2002. View Article : Google Scholar : PubMed/NCBI
20	Lee HY and Hong IS: Double-edged sword of mesenchymal stem cells: Cancer-promoting versus therapeutic potential. Cancer Sci. 108:1939–1946. 2017. View Article : Google Scholar : PubMed/NCBI
21	Trompet S, de Craen AJ, Mooijaart S, Stott DJ, Ford I, Sattar N, Jukema W and Westendorp RG: High innate production capacity of proinflammatory cytokines increases risk for death from cancer: Results of the PROSPER study. Clin Cancer Res. 15:7744–7748. 2009. View Article : Google Scholar : PubMed/NCBI
22	Gocheva V, Chen X, Peters C, Reinheckel T and Joyce JA: Deletion of cathepsin H perturbs angiogenic switching, vascularization and growth of tumors in a mouse model of pancreatic islet cell cancer. Biol Chem. 391:937–945. 2010. View Article : Google Scholar : PubMed/NCBI
23	Dobrzycka B, Mackowiak-Matejczyk B, Terlikowska KM, Kulesza-Bronczyk B, Kinalski M and Terlikowski SJ: Serum levels of IL-6, IL-8 and CRP as prognostic factors in epithelial ovarian cancer. Eur Cytokine Netw. 24:106–113. 2013.PubMed/NCBI
24	Mustea A, Könsgen D, Braicu EI, Pirvulescu C, Sun P, Sofroni D, Lichtenegger W and Sehouli J: Expression of IL-10 in patients with ovarian carcinoma. Anticancer Res. 26:1715–1718. 2006.PubMed/NCBI
25	Fujisawa T, Joshi B, Nakajima A and Puri RK: A novel role of interleukin-13 receptor alpha2 in pancreatic cancer invasion and metastasis. Cancer Res. 69:8678–8685. 2009. View Article : Google Scholar : PubMed/NCBI
26	Charles KA, Kulbe H, Soper R, Escorcio-Correia M, Lawrence T, Schultheis A, Chakravarty P, Thompson RG, Kollias G, Smyth JF, et al: The tumor-promoting actions of TNF-alpha involve TNFR1 and IL-17 in ovarian cancer in mice and humans. J Clin Invest. 119:3011–3023. 2009. View Article : Google Scholar : PubMed/NCBI
27	Zhu W, Shiojima I, Ito Y, Li Z, Ikeda H, Yoshida M, Naito AT, Nishi J, Ueno H, Umezawa A, et al: IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature. 454:345–349. 2008. View Article : Google Scholar : PubMed/NCBI
28	Kumar J, Fraser FW, Riley C, Ahmed N, McCulloch DR and Ward AC: Granulocyte colony-stimulating factor receptor signalling via Janus kinase 2/signal transducer and activator of transcription 3 in ovarian cancer. Br J Cancer. 110:133–145. 2014. View Article : Google Scholar : PubMed/NCBI
29	Lewis AM, Varghese S, Xu H and Alexander HR: Interleukin-1 and cancer progression: The emerging role of interleukin-1 receptor antagonist as a novel therapeutic agent in cancer treatment. J Transl Med. 4:482006. View Article : Google Scholar : PubMed/NCBI
30	Konishi N, Miki C, Yoshida T, Tanaka K, Toiyama Y and Kusunoki M: Interleukin-1 receptor antagonist inhibits the expression of vascular endothelial growth factor in colorectal carcinoma. Oncology. 68:138–145. 2005. View Article : Google Scholar : PubMed/NCBI
31	Owens GL, Price MJ, Cheadle EJ, Hawkins RE, Gilham DE and Edmondson RJ: Ex vivo expanded tumour-infiltrating lymphocytes from ovarian cancer patients release anti-tumour cytokines in response to autologous primary ovarian cancer cells. Cancer Immunol Immunother. 67:1519–1531. 2018. View Article : Google Scholar : PubMed/NCBI
32	Chang CJ, Chen YH, Huang KW, Cheng HW, Chan SF, Tai KF and Hwang LH: Combined GM-CSF and IL-12 gene therapy synergistically suppresses the growth of orthotopic liver tumors. Hepatology. 45:746–754. 2007. View Article : Google Scholar : PubMed/NCBI
33	Su EW, Moore CJ, Suriano S, Johnson CB, Songalia N, Patterson A, Neitzke DJ, Andrijauskaite K, Garrett-Mayer E, Mehrotra S, et al: IL-2Rα mediates temporal regulation of IL-2 signaling and enhances immunotherapy. Sci Transl Med. 7:311ra1702015. View Article : Google Scholar : PubMed/NCBI
34	Broughton SE, Dhagat U, Hercus TR, Nero TL, Grimbaldeston MA, Bonder CS, Lopez AF and Parker MW: The GM-CSF/IL-3/IL-5 cytokine receptor family: From ligand recognition to initiation of signaling. Immunol Rev. 250:277–302. 2012. View Article : Google Scholar : PubMed/NCBI
35	Giuntoli RL II, Webb TJ, Zoso A, Rogers O, Diaz-Montes TP, Bristow RE and Oelke M: Ovarian cancer-associated ascites demonstrates altered immune environment: Implications for antitumor immunity. Anticancer Res. 29:2875–2884. 2009.PubMed/NCBI
36	Gao J, Zhao L, Wan YY and Zhu B: Mechanism of action of IL-7 and its potential applications and limitations in cancer immunotherapy. Int J Mol Sci. 16:10267–10280. 2015. View Article : Google Scholar : PubMed/NCBI
37	Green DS, Nunes AT, Annunziata CM and Zoon KC: Monocyte and interferon based therapy for the treatment of ovarian cancer. Cytokine Growth Factor Rev. 29:109–115. 2016. View Article : Google Scholar : PubMed/NCBI
38	Zsiros E, Duttagupta P, Dangaj D, Li H, Frank R, Garrabrant T, Hagemann IS, Levine BL, June CH, Zhang L, et al: The ovarian cancer chemokine landscape is conducive to homing of vaccine-primed and CD3/CD28-costimulated T cells prepared for adoptive therapy. Clin Cancer Res. 21:2840–2850. 2015. View Article : Google Scholar : PubMed/NCBI
39	Ding L, Li B, Zhao Y, Fu YF, Hu EL, Hu QG, Ni YH and Hou YY: Serum CCL2 and CCL3 as potential biomarkers for the diagnosis of oral squamous cell carcinoma. Tumor Biol. 35:10539–10546. 2014. View Article : Google Scholar
40	Bronger H, Singer J, Windmüller C, Reuning U, Zech D, Delbridge C, Dorn J, Kiechle M, Schmalfeldt B, Schmitt M and Avril S: CXCL9 and CXCL10 predict survival and are regulated by cyclooxygenase inhibition in advanced serous ovarian cancer. Br J Cancer. 115:553–563. 2016. View Article : Google Scholar : PubMed/NCBI
41	Bermudez Y, Aquino MM, Saunders BO, Coppola D, Nicosia SV and Kruk PA: Cytokines modulate telomerase activity in human ovarian cancer cell lines. AACR. 64:2242004.
42	Zhang C, Yang SJ, Wen Q, Zhong JF, Chen XL, Stucky A, Press MF and Zhang X: Human-derived normal mesenchymal stem/stromal cells in anticancer therapies. J Cancer. 8:85–96. 2017. View Article : Google Scholar : PubMed/NCBI
43	Gauthaman K, Fong CY, Subramanian A, Biswas A and Bongso A: ROCK inhibitor Y-27632 increases thaw-survival rates and preserves stemness and differentiation potential of human Wharton's jelly stem cells after cryopreservation. Stem Cell Rev. 6:665–676. 2010. View Article : Google Scholar : PubMed/NCBI