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Objectives: Molecular mechanism of carcinogenesis associated with high risk (HR) type HPV is related to the activity of virus oncoproteins E5, E6 and E7. Their task is to bring the cells to a state enabling synthesis of viral DNA, copying viral particles and promotion of uncontrolled cell growth. Proliferative factors of a feminine genital tract epithelium are also sex hormones — estradiol and progesterone. Steroids influence the transcription of genes involved in cell cycle, acting through specific nuclear receptors (ER and PR). The aim of this study was evaluation of the effect of selected concentrations of 17ß-estradiol and progesterone on survival and proliferation of HeLa line cells. Material and methods: The study was done on a transformed HeLa cell line containing integrated genome HPV of type 18. The lines were incubated in the presence of 17ß-estradiol at the concentrations of lx10-4M, lx10-5M, lx10-7M, lx10-8M. Cell survival was determined with the use of 0.5% of water solution of toluidine blue and the cell proliferation rate were evaluated with the use of BrdU Labelling and Detection Kit and the method ELISA (Roche). Results: The obtained results point to 10% toxic influence on HeLa line cells of 17ß-estradiol at high concentrations after 72 h of incubation. The influence of the hormone on proliferation rate was diversified depending on the hormone concentration and time.
The relationship between HPV (Human Papillomavirus) and cervical carcinoma is very strong, causal, constant, specific and universal. HPV infection precedes the occurrence of preinvasive carcinoma and the evidence for biological probability of such relationship does not leave any doubt. Out of high-risk HPV types, the most often occurring in cervical infections types 16 and 18 were placed in the first group of human carcinogens. However, HPV infection is not the only factor of malignant neoplasm development. Neuroplastic transformation of cervical epithelium requires the presence of cofactors. In recent years, higher role is ascribed to the influences of steroid hormones, including estradiol and progesterone. In normal and neoplastic cells of uterine cervix, the occurrence of receptors for steroid hormones was described. In the regulatory region of oncogenic HPV hormone response elements were found. Receptors combined with steroid hormones functioning as transcription factors may influence the expression of viral oncogens E6 and E7 through stimulation of proliferation and transformation. The aim of this study was to analyse the influence of 17ß-estradiol on the amount of proteins E6 and E7 HPV 18 and to evaluate the transcript E6/E7 after incubation of HeLa line cells with 17ß-estradiol. The results revealed stimulating influence of the hormone on the expression of proteins and mRNA at concentrations of lx10-7M. At high concentrations of 1x10- 4M, estradiol was inhibiting transcription and expression of oncoproteins.
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