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目的探讨5-氮杂-2’-脱氧胞苷酸(5-aza-2’-deoxycitydine,5-Aza-CdR)诱导人乳腺癌MCF-7细胞凋亡及对人类Runt相关转录因子3(Runt-related transcription factor3,RUNX3)基因表达的影响。方法以0.4、1.6、6.4、25.6、102.4μmol/L的5-Aza-CdR分别处理MCF-7细胞,以同体积培养液处理MCF-7细胞作为对照组。MTT法测定不同浓度、不同时间对MCF-7细胞增殖的抑制作用,流式细胞仪检测细胞凋亡,RT-PCR和Western blot法分别检测RUNX3基因mRNA和蛋白的表达,计量资料采用方差分析SNK检验。结果 5-Aza-CdR可抑制MCF-7细胞的生长,其抑制率与药物浓度、作用时间呈依赖关系;5-Aza-CdR作用48h后,流式细胞仪检测0.4～102.4μmol/L5-Aza-CdR实验组细胞凋亡率分别为15.33%、25.35%、27.95%、32.39%、39.27%,与对照组相比差异有统计学意义(P<0.05,0.4μmol/L组除外);RT-PCR和Western-blot检测可见RUNX3 mRNA及RUNX3蛋白表达均增加。结论 5-Aza-CdR诱导人乳腺癌MCF-7细胞凋亡,可能与5-Aza-CdR去甲基化,促进RUNX3基因及蛋白表达有关。
Objective To investigate the apoptosis of human breast cancer MCF-7 cells induced by 5-aza-2’-deoxycitydine (5-Aza-CdR) and its effect on the expression of Runt-related transcription factor 3 -related transcription factor 3, RUNX3) gene expression. Methods MCF-7 cells were treated with 0.4,1.6,6.4,25.6,102.4μmol / L 5-Aza-CdR, respectively. MCF-7 cells were treated with the same volume of culture medium as the control group. MTT assay was used to determine the inhibitory effect of different concentrations and time on the proliferation of MCF-7 cells. The apoptosis of MCF-7 cells was detected by flow cytometry. The expressions of RUNX3 mRNA and protein were detected by RT-PCR and Western blot respectively. test. Results 5-Aza-CdR could inhibit the growth of MCF-7 cells. The inhibition rate was dependent on drug concentration and time. After 48h treatment with 5-Aza-CdR, flow cytometry was used to detect the levels of 0.4 ~ 102.4μmol / The apoptotic rate of CdR group was 15.33%, 25.35%, 27.95%, 32.39%, 39.27%, respectively, which was significantly different from that of control group (P <0.05,0.4μmol / L group) The expression of RUNX3 mRNA and RUNX3 protein were both increased by PCR and Western-blot. Conclusion 5-Aza-CdR induces apoptosis of human breast cancer MCF-7 cells, which may be related to 5-Aza-CdR demethylation and promotion of RUNX3 gene and protein expression.