| Objective:1.To study the effects of folic acid and mir-375 on the biological characteristics of cervical cancer cell Siha;2.Verify the effect of folic acid on the expression of mir-375 in cervical cancer cell Siha;3.Study the effects of folic acid and mir-375 on DNMT1 expression in cervical cancer cell Siha.Methods:1.The cervical cancer cell Siha was cultured with different concentrations of folic acid,and the mi R-375 analog and mi R-375 inhibitor were used to transfect the cervical cancer cell Siha.The invasion and migration ability of the cells in 96 orifice plates,mi R-375 inhibitor,mi R-375 mimics and the control group were observed respectively by the scratch test.Each group had 6 holes,with a total of 18 holes,using Cy.The tation5 cell imaging microplate detection system collects the images of each hole in 0h and 48 h respectively.Through the image J software,the collected images are processed and analyzed.The percentage of Wound Healing per hole at 48 h =(the initial scratch area-the area of a certain time point)/ the initial scratch area is calculated.2.The invasiveness and migration ability of cells in 96 orifice plates containing 500nmol/L,250nmol/L,100nmol/L,50nmol/L,20nmol/L and 0nmol/L were observed by scratch test,with 3 holes in each group,with a total of 18 holes.The same Cytation5 cell imaging microplate detection system was used to collect the images of each hole in 0h and 48 H,respectively.The percentage of Wound Healing per hole in 48 h was calculated by image J software.3.The cervical cancer cell line Siha was cultured with different concentrations of folic acid,and the expression level of mi R-375 was detected by real time fluorescence quantitative PCR technique.The Ct value of each sample was calculated according to the2-??Ctvalue of each group.4.Using a culture medium containing different concen Use of mi R-375 inhibitor,mi R-375 mimics,mi R-375 NC transfection of cervical cancer cells,after 48 hours,the antibody incubation process,such as using Cytation5 cell imaging detection system count every hole collected images,and calculate the control group,mi R-375 inhibitor,mi R-375 mimics group,mi R-375 average intensity of red fluorescence expression in negative control group image(6 holes in each group),on behalf of DNMT1 expression in cervical cancer cells.5.trations of folic acid,the cervical cancer cell line Siha was cultured,and the mi R-375 mimics and mi R-375 inhibitors were used to transfect the cervical cancer cell line Siha.The concentration of folic acid contained in the 96 orifice plates was 0nmol/L,20nmol/L,50nmol/L and 100nmol/L,respectively,by using the Cytation5 cell imaging microplate detection system.Cell images in the medium of 250nmol/L and 500nmol/L,and count the mean red fluorescence intensity in each hole,representing the expression of DNMT1 in cervical cancer cells.Results:1.The mean percentage of wound healing in mir-375 inhibitor group,mir-375 mimics group and control group was(40.6± 5.6)%,(30.4 ±6.3)%,(36.5 ±3.4)%,and there was a significant statistical difference(P<0.05).2.Six groups of different concentrations of folic acid medium the mean percentage of wound healing in cervical cancer cell lines,respectively(20.9±4.7)%,(30.72±3.58)%,(32.5±2.5)%,(34.4±5.4)%,(38.2±7.2)%,(45.2±5.2)%,as a result,the differences were statistically significant(P < 0.05).3.Different concentrations of folic acid medium cervical cancer cell lines Siha,as calculated as 6 groups of sample 2-??Ctaverage respectively(1.0 ±0),(4.5±0.1),(8.6±0.4),(16.8±1.2),(18.6±2.0),(20.3±1.7),the comparison between the two groups,each had statistical significance(P < 0.05).4.Control group,mi R-375 inhibitor,mi R-375 mimics group,mi R-375 negative control group image intensity of red fluorescence expression in average(6 holes in each group),respectively(28465±3221),(29462±4578),(27641±2292),(28739±2374),the comparison between every two groups,suggests in addition to the control group,with no statistical difference between mi R-375 in the negative control group,the rest any comparison between the two groups were statistically significant differences(P < 0.05).5.6 groups of folic acid concentrations in the cell images of 0nmol/L,20nmol/L,50nmol/L,100nmol/L,250nmol/L,500nmol/L,respectively,in the cell images of each cell in the cell images of each hole(3 holes for each group),respectively(25884±1134),(26773±2145),(28968±968),(29625±1280),(32114±2007),(35568±666).There was significant difference between every two groups(P<0.05).Conclusion:1.The proliferation and migration ability of Siha in cervical cancer cells could be inhibited by the use of mi R-375 inhibitor,mi R-375 mimics and Siha of cervical cancer cells using mi R-375 inhibitor and mi R-375 mimics cells with different folic acid concentrations.2.Folic acid can promote the expression of mi R-375 in cervical cancer cell Siha in vitro.3.Folic acid and mir-375 can inhibit the expression of DNMT1 in cervical cancer cell Siha. |
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