CAJ | 학술논문

目的探讨DNA甲基转移酶3b(DNMT3b)在人肝癌细胞株(SMMC7721)中对STAT1和c-myc基因表达及其启动子甲基化水平的影响,并进一步探讨DNMT3b的作用.方法用DNMT3bsiRNA抑制DNMT3b在SMMC7721细胞系中的表达,用Western blot技术检测其转染前后DNMT3b及STAT1和c-myc基因的表达.应用甲基化特异性PCR(MSP)技术分别检测两组细胞中STAT1和c-myc基因启动子区的甲基化状况.结果转染DNMT3bsiRNA的实验组DNMT3b表达水平明显低于对照组,STAT1基因的表达高于对照组;c-myc基因的表达低于对照组.两组中STAT1和c-myc基因启动子区甲基化状态无差异,均未发生甲基化.结论 DNM33b可以调节STAT1和c-myc基因的表达,而不改变基因的甲基化状态,可能发挥了转录调控因子的作用.
목적 탐토DNA갑기전이매3b(DNMT3b)재인간암세포주(SMMC7721)중대STAT1화c-myc기인표체급기계동자갑기화수평적영향,병진일보탐토DNMT3b적작용.방법 용DNMT3bsiRNA억제DNMT3b재SMMC7721세포계중적표체,용Western blot기술검측기전염전후DNMT3b급STAT1화c-myc기인적표체.응용갑기화특이성PCR(MSP)기술분별검측량조세포중STAT1화c-myc기인계동자구적갑기화상황.결과 전염DNMT3bsiRNA적실험조DNMT3b표체수평명현저우대조조,STAT1기인적표체고우대조조;c-myc기인적표체저우대조조.량조중STAT1화c-myc기인계동자구갑기화상태무차이,균미발생갑기화.결론 DNM33b가이조절STAT1화c-myc기인적표체,이불개변기인적갑기화상태,가능발휘료전록조공인자적작용.
Objective To invetigate the influence of DNMT3b of SMMC7721 on the expression of STAT1 and c-myc and the methylation of promoters,and further study the functions of DNMT3b.Methods DNMT3b was silenced by siRNA in human hepatocellular carcinoma cell line SMMC-7721.Westem blotting was performed to evaluate the expression of sTAT1 and c-myc.Methylation specific PCR(MSP)was performed to investigate whether the promoters of STAT1 and c-myc were methylated.Results Western blot analysis showed that the expression of DNMT3b and c-myc in DNMT3bsiRNA transfection group was decreased significantly as compared with the control group,and the expression of STATI increased significanfly.There was no significant difierence in the state of methylation between the transfection and control groups.Conclusion DNMT3b may regulate the expression STAT1 and c-myc in SMMC7721 cells,but not change the state of methylation,suggesting DNMT3b may play roles as transcription factors.