INVOLVEMENT OF HUMAN BETA-DEFENSIN-2 IN PROLIFERATION OF TRANSFORMED CELLS OF HUMAN CERVIX

Markeeva N.V.1, Lysovskiy I.2, Zhuravel E.3, Soldatkina M.3, Lyzogubov V.4, Usenko V.5, Potapov V.6, Pogrebnoy P.V.3

Summary. Aim: To evaluate the influence of human beta-defensin-2 (hBD-2) on viability and proliferation of cultured human epithelial cells and the patterns of hBD-2 expression in normal tissues and early-stage human cervical neoplasia in the relation to proliferative state of these cells. Materials and Methods: The influence of recombinant hBD-2 on viability and proliferation of cultured cells of A431 and M-HeLa lines in vitro was performed by MTT-test, 3H-thymidine incorporation and cell counting techniques. Immunohistochemical analysis of expression of hBD-2 and PCNA in tissue samples (10 normal cases (control), 30 carcinomas of the cervix uteri: 15 – squamous cell carcinoma in situ (Stage 0), and 15 squamous cell carcinoma (Stage Ia)) was performed with the use of anti-hBD-2 and anti-PCNA-mAbs, respectively. Results: We have revealed that hBD-2 significantly stimulated proliferation of A431 and M-HeLa cells in a concentration-dependent manner in the range of 0.1-2 µg/ml, whilst at higher concentrations (> 3-5 µg/ml) it negatively influenced cell viability. The results of immunohistochemical study have shown that malignant transformation of human cervical epithelium is accompanied by the increase of expression of hBD-2 and PCNA. However, the correlative analysis of the expression of the mentioned markers has revealed no relation between them. Conclusion: The effect of hBD-2 on viability and proliferation of cultured epithelial cells possesses a concentration-dependent character. Expression of hBD-2 is increased in early-stage cervical carcinoma.

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