The Anti-Tumor Activities of Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) and Recombinant Fas Molecules in Breast Cancer Tumor Microenvironment

ABSTRACT

Background: Breast cancer, the most common malignancy in women, is a major health problem that is characterized by a defect in cell death (apoptosis). TRAIL and Fas are members of TNF super family that can induce apoptosis and kill malignant cells as wells as bacterial and viral infected cells through activating both apoptotic extrinsic and intrinsic apoptotic pathways. As evasion of apoptosis is thought to be a mechanism of tumor escape, we aimed at the current study to investigate the apoptotic activities of TRAIL and recombinant Fas molecules in breast cancer tumor microenvironment.

Methods:  Brest tumor/normal tissue samples were collected from 30 radically mestectomized breast cancer patients and cultured individually in absence and presence of either TRAIL or rFas molecules. Apoptosis level was measured immune-histochemicaly according to caspase3 staining reaction.

Results: Our results revealed that there is a significant increase in the level of induced apoptosis within the breast tumor tissue cultured with recombinant TRAIL molecules (TT ) than that induced in those cultured with recombinant Fas molecules (TF) or in absence of either (Mean ranks 2.8, 2.2 and 1.1 respectively, p<0.0001).  No significant increase in induced apoptosis levels observed within the normal tissue culture systems N, NF and NT (mean rank 1.8,2 and 2.2 respectively, p = 0.396) while there was a significant higher levels of induced apoptosis observed in the treated tumor culture systems than that in corresponding normal ones (TF vs. NF and TT vs. NT, P<0.001).

Conclusion: TRAIL and Fas have a significant value in selective induction of apoptosis for breast cancer cells within the TME with the superiority of recombinant TRAIL.

Key words:  Breast cancer, tumor microenvironment, Apoptosis, TRAIL, Fas.

REFERENCES

1. Tao Z, Shi A, Lu C, Song T, Zhang Z, Zhao J. Breast Cancer: Epidemiology and Etiology. Cell Biochem Biophys 2015;72(2):333–8.
2. Gerl R. Apoptosis in the development and treatment of cancer. Carcinogenesis 2005;26(2):263–70.
3. Lavrik IN, Krammer PH. Regulation of CD95/Fas signaling at the DISC. Cell Death Differ 2012;19(1):36–41.
4. Johnstone RW, Frew AJ, Smyth MJ. The TRAIL apoptotic pathway in cancer onset, progression and therapy. Nat Rev Cancer 2008;8(10):782–98.
5. Krammer PH. CD95’s deadly mission in the immune system. Nature 2000;407(6805):789–95.
6. Strasser A, Jost PJ, Nagata S. The Many Roles of FAS Receptor Signaling in the Immune System. Immunity2009;30(2):180–92.
7. Wong RSY. Apoptosis in cancer: from pathogenesis to treatment. J Exp Clin Cancer Res 2011;30(1):87.
8. Igney FH, Behrens CK, Krammer PH. CD95L mediates tumor counterattack in vitro but induces neutrophil-independent tumor rejection in vivo. Int J Cancer 2005;113(1):78–87.
9.  Abou-shousha S, Moaz M, Sheta M, Motawea MA. An Approach to Breast Cancer Immunotherapy: The Apoptotic Activity of Recombinant Anti-Interleukin-6 Monoclonal Antibodies in Intact Tumor Microenvironment of Breast Carcinoma . Scand J Immunol 2016;83:427–37.
10. Rahman M, Davis SR, Pumphrey JG, Bao J, Nau MM, Meltzer PS, et al. TRAIL induces apoptosis in triple-negative breast cancer cells with a mesenchymal phenotype. Breast Cancer Res Treat 2009;113(2):217–30.
11. Sateesh Pujari, & Estari Mamidala. (2015). Anti-diabetic activity of Physagulin-F isolated from Physalis angulata fruits. The American Journal of Science and Medical Research, 1(2), 53–60. https://doi.org/10.5281/zenodo.7352308
12. Siegmund D, Wicovsky A, Schmitz I, Schulze-Osthoff K, Kreuz S, Leverkus M, et al. Death receptor-induced signaling pathways are differentially regulated by gamma interferon upstream of caspase 8 processing. Mol Cell Biol 2005;25(15):6363–79.
13. Diao Z, Shi J, Zhu J, Yuan H, Ru Q, Liu S, et al. TRAIL suppresses tumor growth in mice by inducing tumor-infiltrating CD4+CD25+ Treg apoptosis. Cancer Immunol Immunother 2013;62(4):653–63.
14. Kagi D, Vignaux F, Ledermann B, Burki K, Depraetere V, Nagata S, et al. Fas and perforin pathways as major mechanisms of T cell-mediated cytotoxicity. Science (80- ) 1994;265(5171):528–30.
15. Ogasawara J, Watanabe-Fukunaga R, Adachi M, Matsuzawa A, Kasugai T, Kitamura Y, et al. Lethal effect of the anti-Fas antibody in mice. Nature 1993;364(6440):806–9.
16. Kim R, Emi M, Tanabe K, Uchida Y, Toge T. The role of Fas ligand and transforming growth factor beta in tumor progression: molecular mechanisms of immune privilege via Fas-mediated apoptosis and potential targets for cancer therapy. Cancer 2004;100(11):2281–91.
17. Barnhart BC, Legembre P, Pietras E, Bubici C, Franzoso G, Peter ME. CD95 ligand induces motility and invasiveness of apoptosis-resistant tumor cells. EMBO J 2004;23(15):3175–85.
18. Igney FH, Behrens CK, Krammer PH. CD95L mediates tumor counterattack in vitro but induces neutrophil-independent tumor rejection in vivo. Int J Cancer 2005;113(1):78–87.
19. Zhang L, Fang B. Mechanisms of resistance to TRAIL-induced apoptosis in cancer. Cancer Gene Ther 2005;12(3):228–37.
20. Tuettenberg J, Seiz M, Debatin K-M, Hollburg W, von Staden M, Thiemann M, et al. Pharmacokinetics, pharmacodynamics, safety and tolerability of APG101, a CD95-Fc fusion protein, in healthy volunteers and two glioma patients. Int. Immunopharmacol.2012;13(1):93–100.