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目的探讨慢性粒细胞白血病细胞中影响miR-196b表达水平的表观遗传学因素。方法分别采用DNA甲基化转移酶抑制剂5-氮杂-2’-脱氧胞苷(5-Aza-2’-dc)、组蛋白去乙酰化酶抑制剂4-苯基丁酸(PBA)和两者联合处理K562细胞,采用实时定量PCR(Real-time PCR)检测miR-196b的表达水平变化。结果 PBA的半数抑制浓度为1.58mmol/L。和正常人骨髓细胞miR-196b的表达水平相比,Aza组、PBA组和阴性对照组miR-196b的表达水平显著降低且基本一致,Aza+PBA组miR-196b的表达水平和正常人表达水平基本一致。结论单独使用5-Aza-2’-dc或PBA不能使K562细胞中miR-196b的表达恢复正常,两者联合使用共同处理K562细胞,可使miR-196b的表达恢复正常,表明K562细胞中miR-196b的表达水平和基因组甲基化及组蛋白乙酰化均有关系。
Objective To investigate the epigenetic factors affecting the expression of miR-196b in chronic myeloid leukemia cells. Methods DNA methyltransferase inhibitors 5-Aza-2’-deoxycytidine (5-Aza-2’-dc), histone deacetylase inhibitors 4-phenylbutyric acid K562 cells were treated with both of them, and the expression level of miR-196b was detected by real-time PCR. Results The median inhibitory concentration of PBA was 1.58 mmol / L. The expression of miR-196b in Aza group, PBA group and negative control group was significantly lower than that in normal bone marrow cells, and the expression level of miR-196b in Aza + PBA group was significantly lower than that in normal control group Basically the same. CONCLUSION: 5-Aza-2’-dc or PBA alone can not restore the normal expression of miR-196b in K562 cells. The combination of the 5-Aza-2’-dc and PBA can restore the expression of miR-196b in K562 cells. The expression of -196b is related to genomic methylation and histone acetylation.