The Influence Of Estradiol And Estrogen Receptor Inhibitor On The Epithelial-mesenchymal Transition Of Epithelial Ovarian Cancer SKOV3 Cell

Objective:By testing of estrogen and its receptor inhibitor on in vitro cultured ovarian cancer cell line SKOV3 signaling pathways and the transformation of epithelial mesenchymal related protein expression, the influence of to explore the role of estrogen in ovarian cancer development mechanism.Methods:Conventional in vitro cultured human ovarian serous papillary cystadenocarcinoma SKOV3 cell lines, divided into four groups of normal control group, E2 group, ICI182,780 group,E2 + ICI182,780 group. Methods using MTT Colorimetry to detect the conditions of cell proliferation; Annexin V/PI cytometry method to detect cell apoptosis rate; In cultured cells and respectively by different point collection and then Western-Blot method to detect AKT, p-AKT, E-cadherin,Vimentin protein expression; RT-PCR to detect Snail, Slug m RNA transcription factor expression. And using immunofluorescence to detect the ongoing changes of ER-α, ER-β protein expression.Conclusion:1. In vitro culture of SKOV3 cells pseudopodia, began to appear in the estrogen effect after 24 hours cells by oval, spindle mesenchymal cells into polygon sample morphology, intercellular qualitative change significantly over time, the nucleus shrivel.2. E2 action on SKOV3 cell strains after 24 hours and 48 hours, cell apoptosis rates(1.203 ± 0.0764%, 1.99 ± 0.1127%) were significantly lower than control group(2.857 ± 0.3150%, 4.21 ± 0.0173%)(P<0.05). ICI182780 action on SKOV3 cell strains after 24 hours and 48 hours, cell apoptosis rates(5.787 ± 0.8857%, 8.44 ± 1.1530%) were significantly higher than control group(P<0.05).(E2 +ICI182780)group of drugs action on the SKOV3 cell strains after 24 hours and 48 hours, cell apoptosis rates(4.153 ± 0.1457%, 5.98 ± 0.2352%) were higher than the control group, lower than that ICI182780 group(P<0.05).3. E2 can raise SKOV3 cell p-AKT protein expression, and there has time dependent, E2 + ICI182780 combined use to raise p-AKT protein expression level lower than E2 group, ICI182,780 can partially inhibit the above effect of E2, the difference was statistically significant(P<0.05). Three groups of drug are not change AKT protein expression, the difference was not statistically significant(P>0.05).4. E2 can down-regulation of SKOV3 cells E-cadherin protein expression, E2 + ICI182, 780 groups of combined use of down-regulation of E-cadherin protein expression level is less than E2 group, ICI182,780 can partially inhibit E2 to down-regulation the effect of E-cadherin protein(P<0.05). E2 can raise the expression of Vimentin protein of SKOV3 cells, E2 + ICI182, 780 groups of combined use of raised Vimentin protein expression level is less than E2 group, ICI182, 780 can partially inhibit the role of E2 raise Vimentin protein(P<0.05). And E-cadherin expression and Vimentin expression has a negatively correlated between. ICI182,780 group of drugs being applied separately to E-cadherin and Vimentin, and there was no statistically significant difference of two kinds of protein expression(P>0.05).5. E2 action on SKOV3 cells after 24 hours, Snail m RNA, Slug m RNA relative expression levels increased, E2 + ICI182,780 group action on SKOV3 cells after 24 hours, Snail m RNA, Slug m RNA relative expression levels increased. The relative quantitative expression of Snail m RNA, Slug m RNA, the E2 group>E2 + ICI182,780 group>control group(P<0.05). ICI182,780 group being applied separately, the relative quantitative expression difference of Snail m RNA, Slug m RNA was no statistically significant(P>0.05).6. SKOV3 high expression of ER-α,E2 can reduce the protein expression of ER-α, ICI182780 can further reduced the protein expression, the difference was statistically significant(P<0.05).SKOV3 lower expression of ER-β,and ER beta express various dosing group were no significant changes,the difference was no statistically significant(P>0.05).Conclusion:SKOV3 cells and its possible mechanism is E2 by raising p- AKT activation PI3K/AKT signaling pathway, through the classic way to raise the EMT upstream signal transcription factor Snail m RNA, Slug m RNA expression and suppressing the expression of E- cadherin, at the same time increase the stromal cell markers Vimentin protein, induce EMT SKOV3 cells, and estrogen receptor inhibitor ICI182, 780 only partially inhibit the role of E2.