Expression of CD8 marker and proliferation of peripheral blood lymphocyte of multiparous women by Co-culture with breast tumor cell, MDA-231 and MCF-7 lines and comparison with nulliparous women

Document Type : Original Article

Authors

1 MSc Student of Immunology, Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Assistant Professor, Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Professor, Department of Immunology, Reproductive Endocrinology Research Center, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4 Professor, Research Institute for Endocrine Sciences and Metabolism, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Introduction: Low frequency of tumor incidence and metastatic case of breast cancer in multiparous mothers accompanied with this hypothesis that the incidence of microchimerism could be one of the reason of this phenomenon. Therefore, in this study, to investigate mononuclear cells response of peripheral blood adjacent in culture with tumor cell lines of breast cancer, it was attempted to implement an in-vitro co-culture model.
Methods: From 48 multiparous mothers and nulliparous women, Peripheral Blood Mononuclear Cells)PBMC( were purified and isolated by contribution of Ficoll (1077) density gradient centrifugation. By using of trans well system, culture of tumor cell lines (MCF-7 , MDA-231) was performed adjacent with isolated cells. Lymphocyte proliferation assay by BrdU kit and immune-phenotyping of cytotoxic T lymphocyte(CD3+ CD8+) by Flow-cytometric analysis in two period of 48 and 72 h after co-culture was performed.
Results: Mean of lymphocyte proliferation was higher in multiparous women than nulliparous women and was statistically significant in 48 h and 72 h (P<0.001). Also, mean of lymphocyte proliferation of multiparous women in MCF-7 line (P=0.026) and in MDA-231cell line (P<0.001) showed significant results. Determination of the percentage of the population of cytotoxic lymphocytes (CD3 + CD8 +) showed no significant difference between two groups (P=0.603).
Conclusion: The cellular immune responses of multiparous mothers in co-culture with tumor cell lines have clear difference compared with nulliparous women that can lead to the resistance of the mothers against breast tumors. This pheromone could be related to micochimerism during pregnancy. Conclusion about the relation of breast tumor and pregnancy needs to investigate the anti-tumor responses with larger sample size.

Keywords


  1. Sadjadi A, Nouraie M, Mohagheghi MA, Mousavi-Jarrahi A, Malekezadeh R, Parkin DM. Cancer occurrence in Iran in 2002, an international perspective. Asian Pac J Cancer Prev 2005; 6(3):359-63.
  2. Mousavi SM, Gouya MM, Ramazani R, Davanlou M, Hajsadeghi N, Seddighi Z. Cancer incidence and mortality in Iran. Ann Oncol 2009; 20(3):556-63.
  3. Taghavi A, Fazeli Z, Vahedi M, Baghestani AR, Pourhoseingholi A, Barzegar F, et al. Increased trend of breast cancer mortality in Iran. Asian Pac J Cancer Prev 2012; 13(1):367-70.
  4. Gadi V, Nelson JL. Fetal microchimerism in women with breast cancer.Cancer Res2007; 67(19):9035-8.
  5. Stagg J, Johnstone RW, Smyth MJ. From cancer immunosurveillance to cancer immunotherapy. Immunol Rev 2007; 220:82–101.
  6. Mlecnik B, Bindea G, Pages F, Galon J. Tumor immunosurveillance in human cancers. Cancer Metastasis Rev 2011; 30(1):5–12.
  7. Fridman WH, Pages F, Sautes-Fridman C, Galon J. The immune contexture in human tumours: impact on clinical outcome. Nat Rev Cancer 2012; 12(4):298–306.
  8. Denkert C, Loibl S, Noske A, Roller M, Muller BM, Komor M, et al. Tumor-associated lymphocytes as an independent predictor of response to neoadjuvant chemotherapy in breast cancer. J Clin Oncol 2010; 28(1):105–13.
  9. Mahmoud SM, Paish EC, Powe DG, Macmillan RD, Grainge MJ, Lee AH, et al. Tumor-infiltrating CD8+ lymphocytes predict clinical outcome in breast cancer. J Clin Oncol 2011; 29(15):1949–55.
  10. Finak G, Bertos N, Pepin F, Sadekova S, Souleimanova M, Zhao H, et al. Stromal gene expression predicts clinical outcome in breast cancer. Nat Med 2008; 14(5):518–27.
  11. Chen G, Goeddel DV. TNF-R1 signaling: a beautiful pathway. Science 2002; 296(5573):1634–5.
  12. Havell EA, Fiers W, North RJ. The antitumor function of tumor necrosis factor (TNF), I. Therapeutic action of TNF against an established murine sarcoma is indirect, immunologically dependent, and limited by severe toxicity. J Exp Med 1988; 167(3):1067–85.
  13. Balkwill F. TNF-alpha in promotion and progression of cancer. Cancer Metastasis Rev 2006; 25(3):409–16.
  14. Woo CH, Eom YW, Yoo MH, You HJ, Han HJ, Song WK, et al. Tumor necrosis factor-𝛼 generates reactive oxygen species via a cytosolic phospholipase A2-linked cascade. J Biol Chem 2000; 275(41):32357–62.
  15. Hussain SP, Hofseth LJ, Harris CC. Radical causes of cancer. Nature Rev Cancer 2003; 3(4):276–85.
  16. Fatima N, Zaman M, Fatima T. Increased risk of breast cancer in multiparous and lactating women attending a breast care clinic in Pakistan: a paradigm shift? Asian Pac J Cancer Prev 2010; 11(5):1219-23.
  17. Britt K, Ashworth A, Smalley M. Pregnancy and the risk of breast cancer. Endocr Relat Cancer 2007; 14(4):907–33.
  18. Bhadoria AS, Kapil U, Sareen N, Singh P. Reproductive factors and breast cancer: a case–control study in tertiary care hospital of North India. Indian J Cancer 2013; 50(4):316-21.
  19. Fasching PA, Ekici AB, Wachter DL, Hein A, Bayer CM, Häberle L, et al. Breast cancer risk – from genetics to molecular understanding of pathogenesis. Geburtshilfe Frauenheilkd 2011; 71(12):1228-35.
  20. Hsieh C, Pavia M, Lambe M, Lan SJ, Colditz GA, Ekbom A, et al. Dual Effect of parity on breast cancer risk. Eur J Cancer 1994; 30(7):969-73.
  21. Seppanen E, Fisk NM, Khosrotehrani K. Pregnancy-acquired fetal progenitor cells. J Reprod Immunol 2013; 97(1):27-35.
  22. Skowron-Cendrzak A, Rudek Z, Sajak A, Kubera M, Basta-Kaim A, Shani J. Effect of multiparity on T-cell proliferation response to mitogen stimulation in elderly women. Int J Immunopharmacol 1999; 21(3):177-83.
  23. Agrawal B, Reddish MA, Krantz MJ, Longenecker BM. Does pregnancy immunize against breast cancer? Cancer Res 1995; 55(11):2257-61.
  24. Chen L, Xiao Z, Meng Y, Zhao Y, Han J, Su G, et al. The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs. Biomaterial 2012; 33(5):1437-44.