EFFECT OF BRCA1 AFTER UV-B-IRRADIATION IN CELL CYCLE PROGRESSION BETWEEN HUMAN MELANOMA CELL LINES DETERMINED BY FLOW CYTOMETER.
AbstractThe aim of the current study was to evaluated the effect of BRCA1(breast cancer 1 early-0nsent) after UV-B-irradiation in a panel of human malignant melanoma cells using flow cytometry method. Different human melanoma cell lines (SK-MEL28,SK-MEL93, ,SK-MeL93/shBRCA1 and SK-Mel93/shplKO.1) were cultured for triplicaten experiments. Cells were exposed to UV-B (10mJ/cm2)for different point time(1/2,1,3,6,9,12,and 24h). Cells at 0hr were a non-irradiation control. The cell cycle was evaluated by flow cytometry. SK-MEL28 cell line seemed to be synchronized at the G1/S boundary phase until 12h after UVB-irradiation. a decrease in the fraction of S cells and a reciprocal increase of cells in Sub-G1 phase was observed , from 12 to 24h.While, the SK-MEL93 a rapid increase (about 35%) of cells in Sub-G1 phase was observed already 6h after UVB irradiation. Moreover, we found that the SKMel93/shBRCA1 cells seemed an increase (about 15%) of cells in Sub-G1 phase already 12h after UVB irradiation compared with cells infected with the empty lentiviral vector (SK-Mel93/shplKO.1). we conclude that an important role played by BRCA1 providing new insights into the molecular mechanisms underlying UV-induced melanomagenesis.
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