| The effect and the mechanisms of estrogen andprogesterone on proliferation on ovarian cancer cell lines in vitroAbstractObjective : To investigate 17-0 estradiol and progesterone on the effect of cell proliferative, cell cycle kinetics and apoptosis in ovarian cancer cell lines in vitro. And to study the regulatory effect of estradiol and progesterone on expression of the estradiol receptor .progesterone receptor and TGF- a protein in ovarian cancer cell lines.Methods: 1 The proliferation capacity of the ovarian carcinoma cell lines HO8910,PE04 and PEG 14 in the culture medium with 17- & estradiol and progesterone was evaluated by the mircoculture tetrazolium assay (MTT) and cell counter ,and cell cycle was analyzed by flow cytometry. 2 The expression! of the estrogen receptor, progesterone receptor of the cell lines was examined by the immunohistochemical staining . And the expression of ER- a ,ER- & and PRmRNA of the cell lines was examined by the reverse transcrptase - polymerase chain reaction (RT-PCR). 3 The cell apoptosis was detected by flow cytometry , DNA agarose gel electrophoresis and termined deoxynucleotidyl transferase-mediated duTP nick end labeling (TUNEL). 4 The expression of TGF- a protein was analyzed by western blotting. Results : 1 There was not significant difference in proliferation of three celllines was not changed after 24 hours cultured in the medium with 17-P estradiol . After 48 hours ,the growth of HO8910 cell line was inhibited by 17- ?estradiol (1X lO"6-! X10'5 mol/L), and the inhibitive rate of 17- 0 estradiol (1 X 10"M X 10'5 mol/L) was 15%^0%.Compared with the parent cell line ,there was significant difference, PO.05~0.01. 2 Progesterone inhibited the proliferative of three cell lines. After 48 hourstreated with progesterone (1X10'10 mol/L, 1X1(T*~1X10J mol/L), theinhibitive rate of HO8910 cells was 2736%, 14^58%, l+Woand 26r?%,respectively. Progesterone also inhibited the proliferative of PE04 and PE014 after 24 hours , with a dose-dependence manner. The inhibitive rate of PE04 and PE014 treated with progesterone (5 X10"5 mol/L) was 85.75% and 81.2%, respectively. 3 After 48 hours ,the mixture of 17- ?estradiol and progesterone in concentration of 1 X 10"l?mol/L, 1X 10'7~1 X 10"5 mol/L inhibited the proliferative of HO8910 cell lines, and the inhibitive rate was 27.26%, 16.58%, 14.77%and 26.87%,respectively. But the inhibitive rate did not increase after 72 hours. 4 By means of MTT assay ,the result of growth of HO8910 treated with 17- ?estradiol or progesterone in concentration of 1X10"5 mol/L in 21 days inhibited the proliferative of HO8910 cell lines. And the inhibitive rate was 71.51% and 68.04%, respectively. The mixture of 17-0 estradiol and progesterone in concentration of 1X10"5 mol/L in 21 days inhibited the proliferative of HO8910 cell lines too, and the inhibitive rate was 66.77%. 5 By means of cell counter, the test was further used to analysis the effect of PE04 and PE014 ovarian cancer cell lines treated with varying concentration of estrogen or progesterone in 20 days. The result shows that6estrogen-mediated growth effect of PE04 in concentration of 5XIO"5?mol/L was higher than that of control(p<0.05).Comparing with control, itwas increased 39.6% in 5 X 10'5 mol/L and 20% in 1 X 10'5 mol/L respectively. It shows a dose-dependence manner. Varying concentration of estrogen could inhibit the growth of PE014 slightly ,and there was no significance statistically. Varying concentration of progesterone could inhibit the growth of PE04 and PEG 14 cell lines, and it showed a dose-dependence manner.. The result shows that progestrerone-mediated growth effect of cells in concentration of 5 X10"5 mol/L was highest. The inhibitive rate of PE04 treated with progesterone (1-5X10"5 mol/L )was 19.3% and 35.4%,respectively. The inhibitive rate of PE014 treated with progesterone (1-5 X10'5 mol/L )was 28.9% and 37.6%,respectively. 6 Ther were no significant difference in estrogen receptor and progesterone receptor of HO...
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