Suppression of STAT3 by S31-201 to reduce the production of immunoinhibitory cytokines in a HIF1-α-dependent manner: a study on the MCF-7 cell line

  • Amirhossein Jahangiri
  • Maryam Dadmanesh
  • Khodayar GhorbanEmail author


Signal transducer and activator of transcription 3 (STAT3) interacts with many gene promoters and transcription factors such as hypoxia-induced factor 1α (HIF-1α). Recent evidences proposed that STAT3 and HIF-1α together are responsible for angiogenesis and immune response suppression. The main aim of this study was to inhibit STAT3 and HIF-1α and assess their effects on the expression of immunosuppressive cytokines. S31-201 and PX-478 were used to inhibit STAT3 and HIF-1α, respectively. In both hypoxic and normoxic conditions, intracellular levels of HIF-1α were evaluated by western blotting and flow cytometry. Supernatant levels were also measured for VEGF, IL-10, and TGF-β concentration. S31-201 suppressed proliferation of MCF-7 cells and led to reduced HIF-1α expression in both hypoxic and normoxic conditions. It also decreased production of the immunosuppressive cytokines. STAT3 inhibition suppressed tumor cell growth and cytokine production in a HIF-1α-dependent manner, and can be used as a promising target in cancer therapies.


STAT3 HIF-1α S31-201 VEGF IL-10 TGF-β 



The authors thank Gholamreza Valizade and Sara Shirinzade from Iran University of Medical Science for their suggestions and discussion.

Funding information

This study was supported by a grant from Aja Medical University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Adachi M, Cui C, Dodge CT, Bhayani MK, Lai SY (2012) Targeting STAT3 inhibits growth and enhances radiosensitivity in head and neck squamous cell carcinoma. Oral Oncol 48(12):1220–1226CrossRefGoogle Scholar
  2. Chanmee T, Ontong P, Konno K, Itano N (2014) Tumor-associated macrophages as major players in the tumor microenvironment. Cancers 6(3):1670–1690CrossRefGoogle Scholar
  3. Chen L, Shi Y, Yuan J, Han Y, Qin R, Wu Q, Jia B, Wei B, Wei L, Dai G, Jiao S (2014) HIF-1 alpha overexpression correlates with poor overall survival and disease-free survival in gastric cancer patients post-gastrectomy. PLoS One 9(3):e90678CrossRefGoogle Scholar
  4. Courtnay R, Ngo DC, Malik N, Ververis K, Tortorella SM, Karagiannis TC (2015) Cancer metabolism and the Warburg effect: the role of HIF-1 and PI3K. Mol Biol Rep 42(4):841–851CrossRefGoogle Scholar
  5. Favaro E, Lord S, Harris AL, Buffa FM (2011) Gene expression and hypoxia in breast cancer. Genome Med 3(8):55CrossRefGoogle Scholar
  6. Fukada T, Ohtani T, Yoshida Y, Shirogane T, Nishida K, Nakajima K et al (1998) STAT3 orchestrates contradictory signals in cytokine-induced G1 to S cell-cycle transition. EMBO J 17(22):6670–6677CrossRefGoogle Scholar
  7. Gao P, Niu N, Wei T, Tozawa H, Chen X, Zhang C et al (2017) The roles of signal transducer and activator of transcription factor 3 in tumor angiogenesis. Oncotarget 8(40):69139–69161CrossRefGoogle Scholar
  8. Janji B, Viry E, Moussay E, Paggetti J, Arakelian T, Mgrditchian T et al (2016) The multifaceted role of autophagy in tumor evasion from immune surveillance. Oncotarget 7(14):17591–17607CrossRefGoogle Scholar
  9. Klaus A, Fathi O, Tatjana T-W, Bruno N, Oskar K. (2018) Expression of hypoxia-associated protein HIF-1α in follicular thyroid cancer is associated with distant metastasis. Pathol Oncol Res 24(2):289–296CrossRefGoogle Scholar
  10. Mannino MH, Zhu Z, Xiao H, Bai Q, Wakefield MR, Fang Y (2015) The paradoxical role of IL-10 in immunity and cancer. Cancer Lett 367(2):103–107CrossRefGoogle Scholar
  11. Marotta LL, Almendro V, Marusyk A, Shipitsin M, Schemme J, Walker SR et al (2011) The JAK2/STAT3 signaling pathway is required for growth of CD44(+)CD24(−) stem cell-like breast cancer cells in human tumors. J Clin Invest 121(7):2723–2735CrossRefGoogle Scholar
  12. Masoud GN, Li W (2015) HIF-1α pathway: role, regulation and intervention for cancer therapy. Acta Pharm Sin B 5(5):378–389CrossRefGoogle Scholar
  13. Niu G, Briggs J, Deng J, Ma Y, Lee H, Kortylewski M, Kujawski M, Kay H, Cress WD, Jove R, Yu H (2008) Signal transducer and activator of transcription 3 is required for hypoxia-inducible factor-1α RNA expression in both tumor cells and tumor-associated myeloid cells. Mol Cancer Res 6(7):1099–1105CrossRefGoogle Scholar
  14. O'Shea JJ, Murray PJ (2008) Cytokine signaling modules in inflammatory responses. Immunity 28(4):477–487CrossRefGoogle Scholar
  15. Palayoor ST, Mitchell JB, Cerna D, DeGraff W, John-Aryankalayil M, Coleman CN (2008) PX-478, an inhibitor of hypoxia-inducible factor-1α, enhances radiosensitivity of prostate carcinoma cells. Int J Cancer 123(10):2430–2437CrossRefGoogle Scholar
  16. Palomares O, Martin-Fontecha M, Lauener R, Traidl-Hoffmann C, Cavkaytar O, Akdis M et al (2014) Regulatory T cells and immune regulation of allergic diseases: roles of IL-10 and TGF-beta. Genes Immun 15(8):511–520CrossRefGoogle Scholar
  17. Pang M, Ma L, Gong R, Tolbert E, Mao H, Ponnusamy M, Chin YE, Yan H, Dworkin LD, Zhuang S (2010) A novel STAT3 inhibitor, S3I-201, attenuates renal interstitial fibroblast activation and interstitial fibrosis in obstructive nephropathy. Kidney Int 78(3):257–268CrossRefGoogle Scholar
  18. Pawlus MR, Wang L, Hu CJ (2014) STAT3 and HIF1α cooperatively activate HIF1 target genes in MDA-MB-231 and RCC4 cells. Oncogene 33(13):1670–1679CrossRefGoogle Scholar
  19. Schindler C, Levy DE, Decker T (2007) JAK-STAT signaling: from interferons to cytokines. J Biol Chem 282(28):20059–20063CrossRefGoogle Scholar
  20. Seo HS, Ku JM, Choi HS, Woo JK, Jang BH, Shin YC et al (2014) Induction of caspase-dependent apoptosis by apigenin by inhibiting STAT3 signaling in HER2-overexpressing MDA-MB-453 breast cancer cells. Anticancer Res 34(6):2869–2882PubMedGoogle Scholar
  21. Shuai K, Liu B (2003) Regulation of JAK-STAT signalling in the immune system. Nat Rev Immunol 3(11):900–911CrossRefGoogle Scholar
  22. Siddiquee K, Zhang S, Guida WC, Blaskovich MA, Greedy B, Lawrence HR, Yip MLR, Jove R, McLaughlin MM, Lawrence NJ, Sebti SM, Turkson J (2007) Selective chemical probe inhibitor of Stat3, identified through structure-based virtual screening, induces antitumor activity. Proc Natl Acad Sci U S A 104(18):7391–7396CrossRefGoogle Scholar
  23. Sui H, Zhao J, Zhou L, Wen H, Deng W, Li C, Ji Q, Liu X, Feng Y, Chai N, Zhang Q, Cai J, Li Q (2017) Tanshinone IIA inhibits beta-catenin/VEGF-mediated angiogenesis by targeting TGF-beta1 in normoxic and HIF-1alpha in hypoxic microenvironments in human colorectal cancer. Cancer Lett 403:86–97CrossRefGoogle Scholar
  24. Sun X, Zhang J, Hou Z, Han Q, Zhang C, Tian Z (2015) miR-146a is directly regulated by STAT3 in human hepatocellular carcinoma cells and involved in anti-tumor immune suppression. Cell Cycle 14(2):243–252CrossRefGoogle Scholar
  25. Teng Y, Ross JL, Cowell JK (2014) The involvement of JAK-STAT3 in cell motility, invasion, and metastasis. JAKSTAT 3(1):e28086PubMedPubMedCentralGoogle Scholar
  26. Wang Z, Dabrosin C, Yin X, Fuster MM, Arreola A, Rathmell WK, Generali D, Nagaraju GP, el-Rayes B, Ribatti D, Chen YC, Honoki K, Fujii H, Georgakilas AG, Nowsheen S, Amedei A, Niccolai E, Amin A, Ashraf SS, Helferich B, Yang X, Guha G, Bhakta D, Ciriolo MR, Aquilano K, Chen S, Halicka D, Mohammed SI, Azmi AS, Bilsland A, Keith WN, Jensen LD (2015) Broad targeting of angiogenesis for cancer prevention and therapy. Semin Cancer Biol 35:S224–SS43CrossRefGoogle Scholar
  27. Xue C, Xie J, Zhao D, Lin S, Zhou T, Shi S et al (2017) The JAK/STAT3 signalling pathway regulated angiogenesis in an endothelial cell/adipose-derived stromal cell co-culture, 3D gel model. Cell Prolif 50(1):e12307CrossRefGoogle Scholar
  28. Yue FY, Dummer R, Geertsen R, Hofbauer G, Laine E, Manolio S, Burg G (1997) Interleukin-10 is a growth factor for human melanoma cells and down-regulates HLA class-I, HLA class-II and ICAM-1 molecules. Int J Cancer 71(4):630–637CrossRefGoogle Scholar
  29. Yue P, Lopez-Tapia F, Paladino D, Li Y, Chen CH, Namanja AT, Hilliard T, Chen Y, Tius MA, Turkson J (2016) Hydroxamic acid and benzoic acid-based STAT3 inhibitors suppress human glioma and breast cancer phenotypes in vitro and in vivo. Cancer Res 76(3):652–663CrossRefGoogle Scholar
  30. Zhang Y, Zhang L, Zuo Q, Wang Y, Zhang Y, Xu Q, Li B, Chen G (2017) JAK-STAT signaling regulation of chicken embryonic stem cell differentiation into male germ cells. In Vitro Cell Dev Biol Anim 53(8):728–743CrossRefGoogle Scholar
  31. Zheng Y, Ni Y, Huang X, Wang Z, Han WEI (2013) Overexpression of HIF-1α indicates a poor prognosis in tongue carcinoma and may be associated with tumour metastasis. Oncol Lett 5(4):1285–1289CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2018

Authors and Affiliations

  1. 1.Department of Immunology, School of MedicineAja University of Medical SciencesTehranIran
  2. 2.Department of Infectious Diseases, School of MedicineAja University of Medical SciencesTehranIran

Personalised recommendations