Methylation And Demethylation Of MicroRNA-147’s Promoter Region In Glioma

Objective:(1).We collected tumor specimens from 45 patients with different WHO grades of gliomas then detected the methylation of the promoter region of microRNA-147 with MSP and analyzed its relationship with clinicopathological characteristics.(2).We cultured U87 cells in 5’aza-dC medium of different degrees to determine the methylation degree of the promoter region of miR-147 in and cell proliferation status in each group.(3).To determine whether the methylation of the promoter region of miR-147 is related to cell proliferation.Methods:In the first part,methylation-specific polymerase chain reaction(MSP)was used to detect the methylation level of the promoter region of microRNA-147 in the samples collected from glioma patients,and statistical analysis was performed.In the second part,U87 cells were cultured in different concentrations of 5’aza-dC(0 umol/L,4 umol/L and 16umol/L)medium.The 0 umol/L group was used as blank group and the other two groups were used as experimental group.The methylation level of the promoter region of microRNA-147 in each group was detected by Bisulfite Modified Sequencing(BSP)after24 hours.Cell proliferation was observed when cultured cell about 72 hours and was recorded the number of cells.In the third part,Calculate the degree of inhibition of cells in the experimental group.Results:1.Methylation of the promoter region of microRNA-147 was detected by MSP in 45 glioma specimens of different grades.Methylation was found in the promoter region of microRNA-147 in 30 glioma specimens.There was no statistical difference between the methylation and the gender,and age(P > 0.05),but has statistical difference with WHO grades(P<0.05).2.The highest methylation rate was found in the blank group(14.6%),followed by the4umol/L group(8.9%)and the 16umol/L group(3.4%)with the lowest relative cell number.3.About 19.4% of cells were inhibited in 4umol/L group and 51.6% in 16umol/L group.Conclusion:1.The methylation of the promoter region of microRNA-147 in 45 glioma patients was detected.It was proved that the methylation of the promoter region of miR-147 in glioma was not related to the sex and age but related to WHO grade of glioma.2.Methyltransferase inhibitor 5’aza-dC inhibits the methylation of promoter region of microRNA-147,and abnormal expression of methylation in the promoter region of microRNA-147 may be one of the reasons that affect the proliferation of U87 cells,and the lower the degree of methylation,the more obvious the inhibition of cells is.3.Reduced methylation in the promoter region of microRNA-147 may be one of the reasons for the slow proliferation of U87 cells.