Effect Of Folic Acid On E-Cadherin And Vimentin In Cervical Intraepithelial Neoplasia Cells

Objective:Cervical intraepithelial neoplasia(CIN)is a series of precursor lesions closely related to cervical cancer(CC).To study CIN,a stable and good cell model is necessary.The immortalized cervical squamous cell MODEL Ect1/E6E7 carrying HPV16/E6E7 virus can be used as an in vitro cell model for biological studies.To investigate the effects of folic acid on the biological characteristics of Ect1/E6E7 cells and the expression of e-cadherin and Vimentin,markers related to cervical lesions,and then analyze the effects of folic acid on cervical precancerous lesions.Methods:1.Establish CIN cell model in vitro by tissue method and enzyme digestion method,and identify it by immunofluorescence technique;2.Routinely culture cervical immortalized squamous cells Ect1/E6E7,recover the Ect1/E6E7 cell strain,and pass the experiment;3.The concentrations of folic acid in groups of 0,10,100,200,1000 and 2000 nM were used to intervene Ect1/E6E7 cells for 24 h and 48 h,respectively.The inhibition rate of folic acid on Ect1/E6E7 cells was detected by MTT method,and the effect of different concentrations of folic acid on the proliferation of Ect1/E6E7 cells was detected.4.Western Blot was used to detect the expression of E-Cadherin and Vimentin in Ect1/E6E7 cells cultured in groups of 0,10,100,200,1000,2000 nM folate.Results:1.In vitro primary culture experiment of CIN cells was conducted.The primary cell culture lasted a long time,and the cell growth was not ideal.2.MTT method was used to detect the intervention of different concentrations of folic acid on Ect1/E6E7 cells.The results showed that the inhibition rate was statistically significant between the two groups at different intervention times with the same concentration of folic acid(P < 0.05).Under the same intervention time,the inhibition rate was statistically significant between the two groups with different folic acid concentrations(P < 0.05).After 24 h of intervention,Ect1/E6E7 cells proliferated rapidly at a folic acid concentration of 200 nM,and the cell proliferation rate decreased when the concentration was greater than 200 nM.After 48 h of intervention,Ect1/E6E7 cells proliferated rapidly at a folic acid concentration of 200 nM,followed by an increase in the concentration and a decrease in the proliferation rate.3.Western Blot results showed that relative expression levels of Vimentin were 1.097±0.038,0.874±0.04,0.967±0.036,0.856±0.07,0.732±0.032,0.634±0.043,respectively,when folic acid concentration was 0,10,100,200,1000 and 2000 nM,and the relative expression levels of e-cadherin were: Among them,compared with the control group(0nM),the Vimentin expression levels in the folic acid intervention group with different concentrations decreased,and the differences were statistically significant(P values were 0.002,0.013,0.006,0.001,0.001,respectively).Compared with the control group(0nM),the expression level of e-cadherin in the intervention group with different concentrations of folic acid increased,with statistically significant differences(P values of 0.002,0.001,0.011,0.012,0.001,respectively).It was suggested that with the increase of folic acid concentration,the expression of Vimentin in Ect1/E6E7 cells decreased and the expression of e-cadherin increased.Conclusion:The high-concentration folic acid environment inhibited the proliferation of Ect1/E6E7 cells,inhibited the expression of Vimentin in Ect1/E6E7 cells,and promoted the increased expression of E-Cadherin.Folic acid has a certain protective effect on the malignant progression of cervical precancerous cells.