Role Of Decidual Cell Conditioned Media In The Expression Of Invasion-associated Gene Of Ovarian Tumor Cell COC1

1.To study the effect of DCM and its cytokines TNF- a and EGF on the expression of gene associated invasion of ovarian tumor cell COC1 that included MMPs/TIMPs and uPA/PAI-I.2. To explore the molecular mechanism of DCM that interfering MMPs/TIMPs axis and uPA/PAI-I axis of COC1.3. The effect of TNF-? and EGF in DCM were investigated to determine whether they could influence on expression of MMPs/TIMPs and uPA/PAI-I of COC1.METHOD:Trypsin digestion was used to primary culture the human first trimester and third trimester decidual cells. And then DCM was got from primary culture of first trimester and third trimester decidual cells. The immunohistochemical staining was employed to identify purity of decidual cell. Anti-EGF antibody and anti- TNF-? antibody were respectively used to block the function of EGF and TNF- a in DCM. COC1 were respectively treated with DMEM including 15% DCM (control group) , DMEM including 15% DCM and TNF-a antibody at lOng/ml (TNF- a Ab group) and DMEM including 15% DCM and EGF antibody at 10ng/ml(EGF Ab group). Expression level of invasion-associated genes of COC1 was detected by employing the method of reversal transcription polymerase chain reaction. Transwell invasion system was employed to observe the effect of such different substance on invasion capacity of COC1 in vitro.RESULTS:1. MMP-2, TIMP-2 and PAI-1 mRNA were be expressed in COC1, MMP-9, TIMP-1 and uPA mRNA were not. Expression of the MMP-2 of COC1 was apparently down-regulated by first trimester and third trimester DCM compared with that of the controlled group (P<0.01). Expression of the TIMP-2 and PAI-1 mRNA of COC1 were also be apparently up-regulated by both of first trimester and third trimester DCM compared with that of the controlled group (P<0.01).2. MMP-2 mRNA and TIMP-2mRNA were expressed in COC1. MMP-9 mRNA and TIMP-lmRNA were not expressed in COC1. Expression of MMP-2 mRNA of COC1 was significantly up-regulated in TNF- a Ab group (P<0.01) and was not in EGF Ab group (P=0. 28) compared to that of control group. Expression of TIMP-2mRNA of COC1 was significantly down-regulated in TNF-a Ab group P<0.01) and not in EGF Ab group (P>0.05) compared to that of the control group..3. Expression of PAI-1 mRNA of COC1 was significantly up-regulated by TNF-a antibody group compared with that of the controlled group (P<0.01). Such outcome was the same as EGF antibody group (P<0.01).4. Invasion ability of COC1 cultured in vitro was be significantly reduced by the first trimester DCM and third trimester DCM (P<0.05). And the role of first trimester DCM was significantly high compared with that of third trimester DCM (P<0.05). Invasion capacity of COC1 was not significantly enhanced by TNF-a antibody group compared with that of the controlled group DCM (P> 0.05). EGF-Ab group had a trend of debasing invasion capacity of COC1, but not significant compared with that of the controlled group (P > 0.05).CONCLUSION:The decidual cell conditioned media can lower the ability of COCI invasion by interrupting the MMP-2/TIMP-2 axis and uPA /PAI-1 axis of COCI. TNF-a could make role in this mechanism. TNF-a could reduce the ability of DCM that inhibited PA/PAI system of COCI. But EGF could enhance. This outcome show that PA/PAI system could be not dominant invasion system of COCI, but MMPs/TIMPs could be leading system.