BML-111 Inhibits The Migration Of Cervical Cancer Hela Cells By Down-regulating Hypoxia Inducible Factor 1α And 5-lipoxygenase

Objective:Cervical cancer is currently one of the most common gynecological malignant tumors that seriously threatens the health of women around the world.According to previous studies,lipoxygen A4 plays an important role in the development of various tumors.We speculate that the lipoxygen analogues BML-111 may inhibit the migration of cervical cancer cells by regulating hypoxia inducible factor 1αand5-lipoxygenaset,and On this basis,to explore its related mechanism of action and provide experimental basis for finding new treatments for cervical cancer.Methods:1.Immunohistochemistry（IHC）was used to detect the expression of HIF-1αand 5-LOX in 42 cases of cervical cancer（36 cases of squamous cell carcinoma and 6cases of adenocarcinoma）,compared with 20 cases of chronic cervicitis tissue,CD68labeled macrophages and CD31 labeled blood vessels were used to detect the number of macrophages and the density of blood vessels in the two tissues.2.The effect of BML-111 on the survival rate of Hela cells after CoCl₂induction was detected by MTT method.3.Wound Healing assay and transwell experiment were used to detect the effect of BML-111 on the migration ability in CoCl₂-induced Hela cells.4.The effect of BML-111 on the expression of HIF-1αin Hela cells and the localization of HIF-1αin the cells were observed through immunofluorescence experiments.5.Using Western blotting（WB）to detect the effects of BML-111 on the expression of HIF-1α、5-LOX、MMP2 and MMP9 in CoCl₂-induced Hela cells.Results:1.Immunohistochemical results showed that the positive expression rate of HIF-1αin cervical cancer tissue was 71.43%（30/42）,and the positive expression rate in chronic cervicitis tissue was 0%（0/20）.The positive expression rate of 5-LOX in cervical cancer tissue was 78.57%（33/42）,and the positive expression rate in chronic cervicitis tissue was 40%（8/20）.The positive expression rate of CD68 in cervical cancer tissue was 71.43%（30/42）,and the positive expression rate in chronic cervicitis tissue was 15%（3/20）.The expressions of HIF-1α,5-LOX and CD68 in chronic cervicitis tissues were lower than those in cervical cancer tissues（P<0.01）.The number of microvessels in cervical cancer tissue 43.111±10.3230 was significantly higher than that of chronic cervicitis tissue 8.500±1.7795（P<0.01）.2.MTT results:BML-111 has a significant inhibitory effect on the proliferation of Hela cells induced by CoCl₂（P<0.05）.3.Wound Healing assay and transwell experimen results:The effect of BML-111 on the migration ability of Hela cells induced by CoCl₂is as follows:Compared with the control group,the migration ability of Hela cells in CoCl₂group is significantly enhanced（P<0.01）;Compared with the CoCl₂group,the migration ability of Hela cells in the CoCl₂+BML-111 group was significantly reduced（P<0.01）;Compared with CoCl₂+BML-111 group,the migration ability of Hela cells in CoCl₂+BML-111+Boc-2 group was significantly enhanced（P<0.01）.4.Immunofluorescence experiment results:HIF-1αis mainly expressed in the nucleus and cytoplasm.Compared with the control group,the expression of HIF-1αin the CoCl₂group is significantly enhanced,.the expression of HIF-1αwas shown decreased after BML-111,however,Boc-2 could reverse this effect.5.Western blot results:Compared with the blank group,the expression of HIF-1α,5-LOX,MMP2 and MMP9 of Hela cells in CoCl₂group were higher（P<0.05）.Compared with the CoCl₂group,the expression of HIF-1α,5-LOX,MMP2,and MMP9 expression of Hela cells in CoCl₂+BML-111 group were reduced（P<0.05）..Compared with the CoCl₂+BML-111 group,the expression of HIF-1α,5-LOX,MMP2 and MMP9 of Hela cells in CoCl₂+BML-111+Boc-2 group were were increased（P<0.05）.Conclusion:1.BML-111 can inhibit the migration ability of CoCl₂-induced Hela cells.2.BML-111 can inhibit the migration ability of Hela cells by down-regulating hypoxia inducible factor 1αand 5-lipoxygenase.