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目的研究DNA甲基转移酶抑制剂5-杂氮-2′-脱氧胞苷(5-Aza-CdR)对人多发性骨髓瘤细胞株RPMI8226生物学活性以及SOCS-1基因表达的影响。方法不同浓度5-Aza-CdR(0.1、0.5、1.0、2.0、5.0μmol/L)对RPMI8226细胞干预后,采用MTT法检测细胞增殖活性;流式细胞术分析细胞凋亡及细胞周期的改变;Real-timePCR法检测各组细胞SOCS-1mRNA的表达。结果5-Aza-CdR对RPMI8226细胞的生长抑制有明显的时间和剂量依赖性(P<0.05);流式检测结果显示,0.1、0.5、1.0、2.0、5.0μmol/L5-Aza-CdR作用于RPMI8226细胞72h后,细胞凋亡率分别为(29.62±2.87)%、(39.98±2.53)%、(49.07±3.51)%、(60.15±4.54)%和(69.88±3.49)%,且呈剂量依赖性(P<0.05);与对照组相比,5-Aza-CdR处理RPMI8226细胞72h后,细胞阻滞于G0/G1期;Real-timePCR结果显示,SOCS-1基因在RPMI8226细胞中微弱表达,5-Aza-CdR作用后,SOCS-1mRNA表达水平呈剂量依赖性逐渐升高(P<0.05)。结论5-Aza-CdR显著抑制多发性骨髓瘤细胞RPMI8226增殖,可能与诱导SOCS-1基因去甲基化和SOCS-1再表达有关。
Objective To investigate the effects of 5-Aza-2’-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, on the biological activity of human multiple myeloma RPMI8226 cells and the expression of SOCS-1. Methods The proliferation of RPMI8226 cells was induced by different concentrations of 5-Aza-CdR (0.1,0.5,1.0,2.0 and 5.0 μmol / L), and the cell proliferation was detected by MTT assay. The apoptosis and cell cycle were analyzed by flow cytometry. Real-time PCR was used to detect the expression of SOCS-1 mRNA in each group. Results 5-Aza-CdR significantly inhibited the growth of RPMI8226 cells in a time-and dose-dependent manner (P <0.05). Flow cytometry showed that 0.1,0.5,1.0,2.0,5.0μmol / L 5-Aza- The apoptotic rates of RPMI8226 cells after 72 h were (29.62 ± 2.87)%, (39.98 ± 2.53)%, (49.07 ± 3.51)%, (60.15 ± 4.54)% and (69.88 ± 3.49)% respectively, (P <0.05). Compared with the control group, RPMI8226 cells were treated with 5-Aza-CdR for 72h and then blocked in G0 / G1 phase. Real-time PCR results showed that SOCS-1 gene was weakly expressed in RPMI8226 cells, After 5-Aza-CdR treatment, the expression of SOCS-1 mRNA increased gradually (P <0.05) in a dose-dependent manner. Conclusions 5-Aza-CdR significantly inhibits the proliferation of multiple myeloma RPMI8226 cells, which may be related to the demethylation of SOCS-1 gene and the re-expression of SOCS-1.