| Objective:Human papilloma virus(HPV) in cervical chronic inflammation caused by persistent infection is the most important reason of cervical cancer. Lipoxins(lipoxins, LXs) is "brake signal" of inflammatory response [1,2]. It is speculated that lipoxin analogs(BML-111) may inhibit chronic inflammation,which also suppress the occurrence of cervical cancer. The experiment proposed by lipopolysaccharide(LPS) in vitro inflammatory microenvironment, which is to explore the effect of BML-111 on the proliferation and migration in LPS-induced cervical cancer(Hela cell). And compared with the OPN gene silencing Hela cells group, the effect of proliferation and migration and invasion caused by BML-111,which was observed on LPS-induced Hela cell. The aim is to provide experimental evidence for the clinical treatment of cervical cancer. Methods:(1) Using MTT to detect the effect of viability caused by BML-111 on LPS-induced Hela cell;(2) Using Western blot to detect the effect of BML-111 on the expression of OPN, P53, MMP2 and MMP 9 protein in LPS-induced Hela cells;(3) By transfecting Sh RNA-OPN in human Hela cells,the expression of OPN was inhibited. Using Western blot method again to detect the effect of BML-111 on the expression of OPN, P53, MMP2 and MMP 9 protein in LPS-induced Hela cells;(4) Through scratches and Transwell experiments, we observed the the effect of BML-111 on the migration in LPS-induced Hela cells; Results :(1) MTT analysis showed: at 24 h and 48 h in vitro, We are supposed to use different concentrations of LPS to stimulate Hela cells.As the control group, 100,10,1,0.1μg/m L LPS stimulation group OD values were 0.8319 ± 0.01894,1.1611 ± 0.01548,1.1587 ± 0.01805,1.1486 ± 0.01209 and 1.1502 ± 0.01653 respectively, This four concentrations of LPS could promote the proliferation of Hela cells(P<0.05); but there was no significant difference(P value among groups with different concentrations> 0.05).(2) MTT showed, 200μg/L BML-111 was used to detect the proliferation on LPS-induced hela cells.As a result, the control group, LPS group, LPS + BML-111 group, LPS + BML-111 + Boc-2 group,which OD values were 0.8314 ± 0.01462,1.1744 ± 0.01667,0.8338 ± 0.01867 and 1.0344 ± 0.02506 respectively. BML-111 significantly inhibited the proliferation of LPS-induced hela cell(P <0.05). After pretreating with Boc-2, the inhibition effect of BML-111 was disappeared(P <0.05).(3) Western blot showed that in non-transfected group, LPS stimulation group OPN, P53, MMP2 and MMP9 expression was significantly increased(P = 0.000 <0.05), BML-111 significantly inhibited the expression of OPN, P53, MMP2, MMP9 protein in LPS-induced hela cells(P = 0.000 <0.05). And after adding Boc-2 Pretreatment, the inhibitory effect of BML-111 disappeared(OPN P = 0.000, P53 P = 0.005 <0.05).(4) Western blot showed that in transfected Sh RNA-OPN Hela cells of each group, 48 hours after transfection, the control group, LPS group, LPS + BML-111 group and the LPS + BML- 111 + Boc-2 group, each of these groups has no significant difference in the expression of OPN, P53, MMP2 and MMP9 Protein.(5) The scratch test and Transwell method results show that in non-transfected group, related to the control group, migration and invasion ability of LPS stimulation group Hela cells was significantly enhanced(P<0.05); with respect to LPS stimulation group, in LPS + BML-111 group, migration and invasion capacity of hela cell was significantly reduced(P<0.05); related to LPS + BML-111 group, in LPS + BML-111 + Boc-2 group, migration and invasion capacity of hela cell was significantly enhanced(P<0.05). However, in transfected groups, there were no differences in all groups. Conclusion:1.BML-111 significantly inhibited the LPS-induced Hela cell proliferation and migration.2. BML-111 inhibited the proliferation and migration of LPS-induced Hela cell by inhibiting the expression of OPN. |
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