ANALYSIS OF APOPTOSIS FACTORS IN PATIENTS WITH PREINVASIVE AND INVASIVE CERVICAL CANCER

This article assesses the disturbance degree of apoptotic program in patients with preinvasive and invasive cervical cancer by investigating the expression level of genes of caspases-3, -6, -8 и -9 in mononuclear cells of periphery blood and in tumor tissue on the two regulating levels – on mRNA level (transcriptional) and proteolytic activity (post transcriptional). 75 patients with stage III of cervical intraepithelial neoplasias (CIN) III  (middle age of 32,9 ± 7,4),45 patients with stage IA (31,3 ± 6,0), 21 – with stage II (43,6 ± 13,2), 15 – with stage III–IV (46,9 ± 11,1) have been examined. The control group has been formed from 30 almost healthy donors without any cervical pathology and papilloma human virus (control 1) and 30 patients with a preinvasive and microinvasive cervical cancer (control 2). It has been found that in proportion of progress of cervix cancer, the membranous expression of CD95 increases in MPB – fraction (peripheral blood monocytes) when the a CIN and initial stages of cervix cancer, more than two times. Herewith number CD95+-lymphocytes is positively correlated with stage of cervical cancer (r = 0,91; R2  = 0,82; p << 0,01). It has been found out that the activity gain of caspase-8 (r = 0,92; R2  = 0,86; p << 0,01), caspase-6 (r = 0,77; R2 = 0,59; p << 0,01) and reduction activity of caspase-9 (r = –0,60;  R2 = 0,36; p < 0,01) in mononuclear cells of peripheral blood pointed out on the sensitivity increase to Fas-induced apoptosis. Opposite, in tumor tissue, beginning from CIN stage III, apoptosis-resistant phenotype is formed, it were defined by the expression of caspase-3 (r = –0,72; R2 = 0,52, p < 0,01), caspase-6 (r = –0,59; R2 = 0, 38; p < 0,01) и caspase-9 (r = –0,67; R2 = 0,45; p < 0,01) by mRNA level and proteolytic activity. It has been shown, that the cervical cancer development is accompanied by multilateral disturbances of apoptotic processes, which are realized in decreased function of caspases and multilateral resistance of tumor cells in against signals of apoptosis. The obtained results give an opportunity to examine the genes of caspase as a probable prospective biomarkers in complex diagnostic of a CIN and early diagnosis of a cervical cancer (in combination with morphological criteria and other molecular markers of viral origin)

1. Бахидзе Е.В., Лавринович О.Е., Чепик О.Ф., Киселев О.И. Роль вируса папилломы че- ловека в лимфогенном метастазировании плоскоклеточного рака шейки матки. Вопросы онкологии 2011;57(3):318–21. [Bakhidze E.V., Lavrinovich O.E., Chepik O.F., Kiselev O.I. The role of human papilloma virus in lymphatic cancer spread of squamous cell cancer of the cervix uteri. Voprosy onkologii = Issues in Oncology 2011;57(3):318–21. (In Russ.)].

2. Кадагидзе З.Г., Черткова А.И., Славина Е.Г. и др. Фенотип иммунокомпетентных кле- ток и его значение в противоопухолевом иммунном ответе. Вестник РАМН 2011;12:20–5. [Kadagidze Z.G., Chertkova A.I., Slavina E.G. et al. Phenotype of immunocompetent cells and its importance in anti-tumor immune response. Vestnik RAMN = Bulletin of Russian Academy of Medical Sciences 2011;12:20–5. (In Russ.)].

3. Jayshree R.S., Sreenivas A., Tessy M., Krishna S. Cell intrinsic and extrinsic factors in cervical carcinogenesis. Indian J Med Res 2009;130(3):286–95.

4. Du J., Chen G.G., Vlantis A.C. et al. Resistance to apoptosis of HPV 16-infected laryngeal cancer cells is associated with decreased Bak and increased Bcl-2 expression. Cancer Lett 2004;205(1):81–8.

5. Tungteakkhun S.S., Filippova M., Fodor N., Duerksen-Hughes P.J. The full-length isoform of human papillomavirus 16 E6 and its splice variant E6* bind to different sites on the procaspase 8 death effector domain. J Virol 2010;84(3):1453–63.

6. Krammer P.H., Arnold R., Lavrik I.N. Life and death in peripheral T cells. Nat Rev Immunol 2007;7(7):532–42.

7. Olsson M., Zhivotovsky B. Caspases and cancer. Cell Death Differ 2011;18(9):1441–9.

8. Yuen M.F., Hughes R.D., Heneghan M.A. et al. Expression of Fas antigen (CD95) in peripheral blood lymphocytes and in liver- infiltrating, cytotoxic lymphocytes in patients with hepatocellular carcinoma. Cancer 2001;92(8):2136–41.

9. Dworacki G., Meidenbauer N., Kuss I. et al. Decreased zeta chain expression and apoptosis in CD3+ peripheral blood T lymphocytes of patients with melanoma. Clin Cancer Res 2001;7(3 Suppl):947s–57s.

10. Ma Y.X., Ye F., Chen H.Z. et al. Study of apoptosis and Fas expression of peritoneal fluid and peripheral blood T lymphocytes in patients with epithelial ovarian cancer and their relationship with CA125. Zhonghua Yi Xue Za Zhi 2007;87(11): 734–9.

11. Hoffmann T.K., Dworacki G., Tsukihiro T. et al. Spontaneous apoptosis of circulating T lymphocytes in patients with head and neck cancer and its clinical importance. Clin Cancer Res 2002;8(8):2553–62.

12. Yoshikawa T., Saito H., Osaki T. et al. Elevated Fas expression is related to increased apoptosis of circulating CD8+ T cell in patients with gastric cancer. J Surg Res 2008;148(2):143–51.

13. Hoser G., Wasilewska D., Domagała- Kulawik J. Expression of Fas receptor on peripheral blood lymphocytes from patients with non-small cell lung cancer. Folia Histochem Cytobiol 2004;42(4):249–52.

14. Reesink-Peters N., Hougardy B.M., van den Heuvel F.A. et al. Death receptors and ligands in cervical carcinogenesis: an immunohistochemical study. Gynecol Oncol 2005;96(3):705–13.

15. Aréchaga-Ocampo E., Pereira-Suárez A.L., del Moral-Hernández O. et al. HPV+ cervical carcinomas and cell lines display altered expression of caspases. Gynecol Oncol 2008;108(1):10–8.

16. Ekonomopoulou M.T., Babas E., Mioglou-Kalouptsi E. et al. Changes in activities of caspase-8 and caspase-9 in human cervical malignancy. Int J Gynecol Cancer 2011;21(3):435–8.

17. Wang W., Fang Y., Sima N. et al. Triggering of death receptor apoptotic signaling by human papillomavirus 16 E2 protein in cervical cancer cell lines is mediated by interaction with c- FLIP. Apoptosis 2011;16(1):55–66.

18. Jäger R., Zwacka R.M. The enigmatic roles of caspases in tumor development. Cancers 2010;2(4):1952–79.

19. Ibrahim R., Frederickson H., Parr A. et al. Expression of FasL in squamous cell carcinomas of the cervix and cervical intraepithelial neoplasia and its role in tumor escape mechanism. Cancer 2006;106(5): 1065–77.

20. Patel S., Chiplunkar S. Host immune responses to cervical cancer. Curr Opin Obstet Gynecol 2009;21(1):54–9.

21. de Wilde J., De-Castro Arce J., Snijders P.J. et al. Alterations in AP-1 and AP-1 regulatory genes during HPV-induced carcinogenesis. Cell Oncol 2008;30(1):77–87.

22. Jacobs J.F., Idema A.J., Bol K.F. et al. Prognostic significance and mechanism of Treg infiltration in human brain tumors. J Neuroimmunol 2010;225(1–2): 195–9.

23. Würstle M.L., Laussmann M.A., Rehm M. The caspase-8 dimerization/dissociation balance is a highly potent regulator of caspase-8, -3, -6 signaling. J Biol Chem 2010;285(43):33209–18.

24. Zhou J.H., Chen H.Z., Ye F. et al. Fas- mediated pathway and apoptosis in normal cervix, cervical intraepithelial neoplasia and cervical squamous cancer. Oncol Rep 2006;16(2):307–11.