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目的研究5-杂氮-2’-脱氧胞啶(5aza-CdR)联合各种组蛋白乙酰基转移酶(HDAC)抑制剂对体外人尤文瘤细胞株RD-ES细胞的抑制作用,为尤文瘤的治疗提供实验参考。方法体外培养5×103个/ml的RD-ES细胞株,以MTT比色法观察不同浓度5aza-CdR(终浓度为10、40、100ng/ml)联合HDAC抑制剂(10、100、400、1000ng/ml的MS-275,2.5、5、10ng/ml的TSA,0.6、1、2.5ng/ml的Depsi)对RD-ES细胞染毒48h的生长抑制作用,以高灵敏度DNA试剂盒检测5aza-CdRDNA抑制率。以RT-PCR法检测5aza-CdR联合Depsi染毒对RD-ES细胞ECAD和TSLC1表达的影响。结果5aza-CdR对RD-ES细胞系抑制率为50%时的抑制剂浓度(IC50)为40ng/ml,48hDNA50%抑制率的浓度约为50ng/ml。HDAC抑制剂(MS-275,trichostatin-A,Depsi)对RD-ES细胞系的IC50值分别为400、2.5、0.6ng/ml。5aza-CdR联合HDAC抑制剂染毒组RD-ES细胞抑制率均高于相同HDAC抑制剂单独染毒组,差异有统计学意义(P<0.05)。与5aza-CdR、Depsi联合染毒组比较,5aza-CdR单独染毒组和Depsi单独染毒组激活ECAD和TSCL1的RT-PCR表达较低,差异均有统计学意义(P<0.05)。结论HDAC抑制剂能增强5aza-CdR对RD-ES细胞的抑制作用。Depsi能增强5aza-CdR激活ECAD和TSCL1的表达。
Objective To investigate the inhibitory effect of 5-aza-2’-deoxycytidine (5aza-CdR) combined with various histone acetyltransferase (HDAC) inhibitors on RD-ES cells in vitro, The treatment provides experimental reference. Methods 5 × 103 cells / ml RD-ES cells were cultured in vitro. MTT assay was used to observe the effects of 5-aza-CdR (10,40 and 100 ng / ml) combined with HDAC inhibitor (10,100,400, 1000 ng / ml MS-275, 2.5, 5, 10 ng / ml TSA, 0.6, 1, 2.5 ng / ml Depsi) for 48 h on RD-ES cells. -CdRDNA inhibition rate. The effects of 5aza-CdR combined with Depsi on the expression of ECAD and TSLC1 in RD-ES cells were detected by RT-PCR. Results The inhibitory concentration (IC50) of 5-aza-CdR was 50ng / ml for RD-ES cell line, and 50ng / ml for 48h DNA50% inhibition rate. The IC50 values of HDAC inhibitors (MS-275, trichostatin-A, Depsi) to RD-ES cell lines were 400, 2.5 and 0.6 ng / ml, respectively. The inhibitory rates of RD-ES cells in 5-aza-CdR combined with HDAC inhibitor-treated group were significantly higher than those in the same HDAC inhibitor-treated group (P <0.05). Compared with 5aza-CdR and Depsi group, RT-PCR expression of ECAD and TSCL1 in 5aza-CdR group and Depsi group were lower than that in 5aza-CdR group and Depsi group (P <0.05). Conclusions HDAC inhibitors can enhance the inhibitory effect of 5aza-CdR on RD-ES cells. Depsi enhances 5aza-CdR activation of ECAD and TSCL1 expression.