MiR-375 Methylation And Expression Of 5-Aza-CdR In Pancreatic Beta Cell Lines

Object : Discussion 5-Aza-2’-deoxycytidine on mouse pancreatic β cell line in the mi R-375 gene expression and DNA methylation influences.Methods:(1) Mouse pancreatic β cell line were treated with different dose of 5-Aza-Cd R, negative control group without medicine, 0umol/L group with 0umol/L 5-Aza-Cd R, 0.08umol/L group with 0.08umol/L5-Aza-Cd R, 0.4umol/L group with 0.4umol/L 5-Aza-Cd R, 2umol/L group with 2umol/L 5-Aza-Cd R,10umol/L group with 10umol/L 5-Aza-Cd R, 50umol/L group with 50umol/L 5-Aza-Cd R, with a determined by MTT method to detect drug after 24 h, 48 h, 72 h cell growth inhibition.(2) Using matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS)to detect different concentrations of 5-Aza-Cd R treatment 24 h, 48 h, 72 h of MIN6 cell line and NIH3T3 cell line without drug treatment in mi R-375 gene DNA methylation status.(3) Using q RT-PCR detection of NIH3T3 cells and 5-Aza-Cd R(50umol/L) treated MIN6 cells 24 h, 48 h,72h after the expression of mi R-375.Results:(1) The inhibitory effect of 5-Aza-Cd R on MIN6 cells, 2umol/L 5-Aza-Cd R group at 24 h, 48 h, 72 h and the rest of the group, the concentrations of comparison OD value were significant differences(P < 0.05), and the comparison between the periods the difference was statistically significant(P <0.05).(2) MIN6 cells were treated with different concentrations of 5-Aza-Cd R treatment 24 h, 48 h, 72 h, to detect the expression of mi R-375 gene DNA showed hypermethylation state where 50umol/L group, mi R-375 of the average methylation were lower than other concentrations.(3) MIN6 cells were treated with 50 umol/L group of 5-Aza-Cd R treatment 24 h, 48 h, 72 h after detecting expression 0umol / L group differences in each group mi R-375 was statistically significant(P <0.05), and expression of mi R-375 was higher than 0umol/L group.(4) The expression of NIH3T3 in mi R-375 cell group was higher than that in MIN6 cell group, and the difference was statistically significant(P <0.05).Conclusions:(1) 5-Aza-Cd R can inhibit the proliferation of MIN6 cells.(2) Compared with NIH3T3 cells, mi R-375 gene promoter region of DNA methylation status in MIN6 cells was high, low expression.(3) 50 umol / L concentrations of 5-Aza-Cd R make MIN6 cells mi R-375 gene DNA demethylation, activate the expression of mi R-375, and its methylation and expression was negatively correlated.