Objective:To explor the bioactive drug β-lapachone of quinone oxidoreductase 1(NQO1)on cervical cancer cell proliferation,migration,invasion metabolism and angiogenesis.The role in the process and its possible molecular mechanism provide new directions and basic experimental support for the clinical treatment of cervical cancer.Methods:The expression of NQO1 in different cervical cancer cells and normal cervical epithelial cells was detected by Western blot.The MTT assay was used to detect the effect of β-lapachone(0,1,2,4,6,8 μmol/L)on the proliferation of cervical cancer cells SiHa and C33a after 24 h,48 h,and 72 h.Cervical cancer cells SiHa(0,1,2,4 μmol/L)and C33a(0,2,4,6 μmol/L)were treated with β-lapachone,and then the morphology of cervical cancer cells was observed with an inverted microscope.The effect of β-lapachone on the colony-forming ability of cervical cancer cells SiHa and C33a was detected by plate clone formation test.The scratch test is used to detect the effect of β-lapachone on the lateral migration ability of cervical cancer cells SiHa and C33a.Transwell test was used to detect the effect of(3-lapachone on the ability of longitudinal migration and invasion of cells.Angiogenesis experiments were used to test the angiogenesis ability of human umbilical vein endothelial cells(HUVECs)cultured from the supernatant of cervical cancer cells SiHa and C33a cells treated with β-lapachone.Western blot test to detect vascular growth factor,epithelial-mesenchymal transition(EMT),Phosphatidylinositol 3-kinase/protein kinase/Bmammalian target ofrapamycin(PI3K/AKT/mTOR)signal pathway related protein expression level.Results:Western blot experiments showed that compared with normal cervical epithelial cells,the expression of NQO1 in cervical cancer cells SiHa and C33a cells was higher,and the expression of NQO1 was lower in HeLa cells.The results of MTT assay showed that after different concentrations of β-lapachone treated SiHa and C33a cells for different time,the proliferation ability of the two cervical cancer cells was significantly reduced(all P values<0.05).Observation under an inverted microscope,it was found that SiHa cells changed from long spindle to oval after β-lapachone treatment,and C33a cells changed from multi-antennary long spindle to oval.The results of the cloning experiment showed that compared with the untreated group,the colony formation ability was significantly inhibited in the treated group(all P values<0.01).The results of the scratch test and the Transwell test showed that,compared with the control group,the drug group significantly inhibited the lateral,vertical migration ability and cell invasion ability of the cells(all P values<0.01).Angiogenesis experiments showed that human umbilical vein endothelial cells cultured with β-lapachone treated cervical cancer cell supernatants had significantly lower angiogenic ability than control group(all P values<0.01).The results of Western blotting showed that the expression levels of vascular endothelial growth factor expression level was down-regulated(all P values<0.05),and the expression level of E-cadherin,an epithelial marker of EMT pathway-related proteins,was up-regulated than that of the control group(all P values<0.01),and the expression levels of interstitial markers Vimentin and Snail were down-regulated(all P values<0.01).PI3K/AKT/mTOR signaling pathway-related proteins expressed p-mTOR and p-AKT expression were down-regulated(all P values<0.05).Conclusion:NQO1’s bioactivation drug β-lapachone mediates PI3K/AKT/mTOR signaling pathway in vitro to inhibit cervical cancer cell proliferation,regulate the expression of pathway-related proteins,and inhibits cell migration and invasion through angiogenesis and epithelial-mesenchymal transition pathway. |