The Anti-tumoral Effect of β-D-Mannuronic Acid (M2000) as a Novel NSAID on Treg Cells Frequency and MMP-2, MMP-9, CCL22 and TGFβ1 Gene Expression in Pre-surgical Breast Cancer Patients
With respect to the role of chronic inflammation in the induction and progression of breast cancer (BC). The relationship between tumor and tumor microenvironment may be a hopeful strategy for BC therapy. According to the effect of β-D-Mannuronic acid (M2000) as a novel non-steroidal anti-inflammatory drug (NSAID) on BC murine model and 4T1 cell line, we started to study that was a phase II, randomized, controlled clinical trial. 24 women with BC were included in this study and were followed by fixed oral doses of M2000, 500 mg two times a day (6-8 weeks). Blood samples were collected at baseline and weeks 6-8. To compare the patterns of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), C-C motif chemokine ligand 22 (CCL22) and The transforming growth factor-beta 1 (TGFβ1) gene expression and T regulatory cells (Tregs) frequency of healthy women normal controls with BC patients, a set of 10 blood samples of women healthy volunteers was collected. The gene expression was evaluated by quantitative Real-time PCR (qRT-PCR) and the frequency of Tregs was assessed by flow cytometry. Our results showed, reduction in MMP-2 (p=0.08), MMP-9 (p=0.03), CCL22 (p=0.003) and TGFβ1 (p=0.1) gene expression and Tregs frequency (p=0.01) which play a main role in the development of chronic inflammation, angiogenesis, tumorigenesis and metastasis. Our findings demonstrated that M2000 therapy as a novel designed NSAID had valuable therapeutic effects on BC. No adverse effects were observed following the use of M2000 after 6-8 weeks.
of breast cancer. annu rev public health 1996; 17(1):47-67.
2. Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns
in GLOBOCAN 2012. Int J Cancer 2015; 136(5):E359-E86.
3. Akbari ME, Sayad S, Sayad S, Khayamzadeh M, Shojaee L, Shormeji Z, et al. Breast Cancer Status in Iran: Statistical Analysis of 3010 Cases between 1998 and 2014. Int J Breast Cancer 2017; 2017:248.
4. Owonikoko TK, Arbiser J, Zelnak A, Shu H-KG, Shim H, Robin AM, et al. Current approaches to the treatment of metastatic brain tumours. Nat Rev Clin Oncol 2014; 11(4):203-22.
5. Kazemi T, Younesi V, Jadidi-Niaragh F, Yousefi M. Immunotherapeutic approaches for cancer therapy: an updated review. Artif Cells Nanomed Biotechnol 2016; 44(3):769-79.
6. Whiteside T. The tumor microenvironment and its role in promoting tumor growth. Oncogene 2008; 27(45):5904-12.
7. Mylona E, Nomikos A, Magkou C, Kamberou M, Papassideri I, Keramopoulos A, et al. The clinicopathological and prognostic significance of membrane type 1 matrix metalloproteinase (MT1-MMP) and MMP-9 according to their localization in invasive breast carcinoma. Histopathology 2007; 50(3):338-47.
8. Ranogajec I, Jakić-Razumović J, Puzović V, Gabrilovac J. Prognostic value of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase N/CD13 in breast cancer patients. Med Oncol 2012; 29(2):561-9.
9. Sung H, Choi J-Y, Lee S-A, Lee K-M, Han S, Jeon S, et al. The association between the preoperative serum levels of lipocalin-2 and matrix metalloproteinase-9 (MMP-9) and prognosis of breast cancer. BMC cancer 2012; 12(1):193.
10. Zeng Y, Liu C, Dong B, Li Y, Jiang B, Xu Y, et al. Inverse correlation between Naa10p and MMP-9 expression and the combined prognostic value in breast cancer patients. Med Oncol 2013; 30(2):562.
11. Bierie B, Moses HL. Tumour microenvironment: TGFβ: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer 2006; 6(7):506-20.
12. Neuzillet C, Tijeras-Raballand A, Cohen R, Cros J, Faivre S, Raymond E, et al. Targeting the TGFβ pathway for cancer therapy. Pharmacol Ther 2015; 147:22-31.
13. Zhu Q, Han X, Peng J, Qin H, Wang Y. The role of CXC chemokines and their receptors in the progression and treatment of tumors. J Mol Histol 2012; 43(6):699-713.
14. Jafarzadeh A, Fooladseresht H, Minaee K, Bazrafshani M, Khosravimashizi A, Nemati M, et al. Higher circulating levels of chemokine CCL22 in patients with breast cancer: evaluation of the influences of tumor stage and chemokine gene polymorphism. Tumour Biol 2015; 36(2):1163-71.
15. Franciszkiewicz K, Boissonnas A, Boutet M, Combadière C, Mami-Chouaib F. Role of chemokines and chemokine receptors in shaping the effector phase of the antitumor immune response. Cancer res 2012; 72(24):6325-32.
16. Kim S, Lee A, Lim W, Park S, Cho MS, Koo H, et al. Zonal difference and prognostic significance of foxp3 regulatory T cell infiltration in breast cancer. J breast cancer 2014; 17(1):8-17.
17. Shou J, Zhang Z, Lai Y, Chen Z, Huang J. Worse outcome in breast cancer with higher tumor-infiltrating FOXP3+ Tregs: a systematic review and meta-analysis. BMC cancer 2016; 16(1):687.
18. Mirshafiey A, Rehm B, Abhari RS, Borzooy Z, Sotoude M, Razavi A. Production of M2000 (β-d-mannuronic acid) and its therapeutic effect on experimental nephritis. Environ Toxicol Pharmacol 2007; 24(1):60-6.
19. Mirshafiey A, Cuzzocrea S, Rehm B, Mazzon E, Saadat F, Sotoude M. Treatment of experimental arthritis with M2000, a novel designed non‐steroidal anti‐inflammatory drug. Scand J Immunol 2005; 61(5):435-41.
20. Mirshafiey A, Cuzzocrea S, Rehm B, Matsuo H. M2000: a revolution in pharmacology. Med Sci Monit 2005; 11(8):PI53-PI63.
21. Mirshafiey A, Matsuo H, Nakane S, Rehm BH, Koh C-S, Miyoshi S. Novel immunosuppressive therapy by M2000 in experimental multiple sclerosis. Immunopharmacol Immunotoxicol 2005; 27(2):255-65.
22. Hosseini F, Hassannia H, Mahdian Shakib A, Jadidi Niaragh F, Enderami SE, Fattahi M, et al. Targeting of crosstalk between tumor and tumor microenvironment by β D mannuronic acid (M2000) in murine breast cancer model. Cancer med 2017; 6(3):640-50.
23. Agrawal A, Fentiman I. NSAIDs and breast cancer: a possible prevention and treatment strategy. Int J Clin Pract 2008; 62(3):444-9.
24. Cuzick J, Otto F, Baron JA, Brown PH, Burn J, Greenwald P, et al. Aspirin and non-steroidal anti-inflammatory drugs for cancer prevention: an international consensus statement. lancet oncolo 2009; 10(5):501-7.
25. Khuder S, Mutgi A. Breast cancer and NSAID use: a meta-analysis. Br j cancer 2001; 84(9):1188.
26. Jafarnezhad-Ansariha F, Yekaninejad MS, Jamshidi A-r, Mansouri R, Vojdanian M, Mahmoudi M, et al. The effects of β-d-mannuronic acid (M2000), as a novel NSAID, on COX1 and COX2 activities and gene expression in ankylosing spondylitis patients and the murine monocyte/macrophage, J774 cell line. Inflammopharmacology 2018; 26(2):375-84.
27. Rastegari Pouyani M, Mostafaie A, Mansouri K, Mortazavi-Jahromi SS, Mohammadi‐Motlagh HR, Mirshafiey A. Anti-angiogenesis effect of β-D-mannuronic acid (M2000) as a novel NSAID with immunosuppressive properties under experimental model. Clin Exp Pharmacol Physiol 2018; 45(4):370-6.
28. Lee AH, Happerfield LC, Bobrow LG, Millis RR. Angiogenesis and inflammation in ductal carcinoma in situ of the breast. J Pathol 1997; 181(2):200-6.
29. Lin EY, Li J-F, Gnatovskiy L, Deng Y, Zhu L, Grzesik DA, et al. Macrophages regulate the angiogenic switch in a mouse model of breast cancer. Cancer res. 2006;66(23):11238-46.
30. Klemm F, Joyce JA. Microenvironmental regulation of therapeutic response in cancer. Trends Cell Biol 2015; 25(4):198-213.
31. Farahani MM, Motevaseli E, Maghsood F, Heidari-Kharaji M, Mirshafiey A. Anti-inflammatory property of β-D-mannuronic acid (M2000) on expression and activity of matrix metalloproteinase-2 and-9 through CD147 molecule in phorbol myristate acetate-differentiated THP-1 cells. Iran J Allergy Asthma Immunol 2017; 16(5):443-51.
32. Mirshafiey A, Khorramizadeh MR, Saadat F, Rehm BH. Chemopreventive effect of M2000, a new anti-inflammatory agent. Med Sci Monit 2004; 10(10):PI105-I9.
33. Mirshafiey A, Taeb M, Mortazavi-Jahromi S, Jafarnezhad-Ansariha F, Rehm BH, Esposito E, et al. Introduction of β-d-mannuronic acid (M2000) as a novel NSAID with immunosuppressive property based on COX-1/COX-2 activity and gene expression. Pharmacol rep 2017; 69(5):1067-72.
34. Irjala H, Alanen K, Grénman R, Heikkilä P, Joensuu H, Jalkanen S. Mannose receptor (MR) and common lymphatic endothelial and vascular endothelial receptor (CLEVER)-1 direct the binding of cancer cells to the lymph vessel endothelium. Cancer res 2003; 63(15):4671-6.