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作 者:张晓宇 白杰[1] 康晓宁[2] 靳丽君 王鹏[1] 刘伟[1] 王遵义[1] 李国琪 ZHANG Xiao-yu;BAI Jie;KANG Xiao-ning;JIN Li-jun;WANG Peng;LIU Wei;WANG Zun-yi(Third Department of Oncological Surgery,Heibei 061001, China;Second Department of Ultrasonography, Cangzhou Central Hospital, Heibei 061001, China)
机构地区:[1]河北省沧州市中心医院肿瘤外三科,061001 [2]河北省沧州市中心医院肿瘤超声二科,061001 [3]不详
出 处:《天津医药》2018年第3期234-238,共5页Tianjin Medical Journal
基 金:沧州市科学技术研究与发展指导计划(172302004)
摘 要:目的分析5-氮杂-2′-脱氧胞苷(5-Aza-Cd R)对体外培养的人乳腺癌细胞Hs578T增殖的影响,及其对此细胞中PRDM10基因甲基化状态的影响。方法分别采用浓度为0、1、3、5μmol/L的5-Aza-Cd R处理体外培养的人乳腺癌细胞系Hs578T。MTT实验检测细胞增殖情况,甲基化特异性PCR(MSP)法检测PRDM10的甲基化状态;RT-PCR和Western blot分别检测PRDM10的m RNA和蛋白表达水平。结果 MTT结果显示,5-Aza-Cd R的浓度越高、处理时间越长,对Hs578T细胞增殖的抑制作用越显著。与0μmol/L组比较,1μmol/L组处理72 h后及3、5μmol/L组处理48 h后,Hs578T的增殖受到显著抑制(P<0.05)。MSP结果显示,5-Aza-Cd R的浓度越高,对PRDM10的去甲基化作用越显著。RT-PCR和Western blot结果显示,5-Aza-Cd R的浓度越高,PRDM10的m RNA和蛋白表达水平越高(P<0.05)。结论 5-Aza-Cd R可抑制Hs578T细胞的增殖,可能与该细胞中PRDM10基因的去甲基化作用有关。To investigate effects of5-Aza-2′-deoxycytidine(5-Aza-CdR)on proliferation of human breast cancer cell line Hs578T was cultured with1,3and5μmol/L DNA methylation inhibitor5-Aza-CdR respectively,and untreated cells were used as control.Cell proliferation was detected by MTT assay.Methylation-Specific PCR(MSP)wasused to detect the methylation status of PRDM10gene.The mRNA and protein expression levels of PRDM10gene weredetected by RT-PCR and Western blot assay.Results MTT results showed that the higher the concentration of5-Aza-CdR,and the longer the treatment time,the more significant inhibitory effect on the proliferation of Hs578T cells.Comparedwith the control group(0μmol/L),the proliferation of Hs578T was significantly inhibited after the treatment for72h in the1μmol/L group,and for48h in the3μmol/L and5μmol/L groups(P<0.05).MSP results showed that the higher theconcentration of5-Aza-CdR,the more significant demethylation of PRDM10.Results of RT-PCR and Western blot showedthat the higher the concentration of5-Aza-CdR,the higher the expression levels of mRNA and protein in PRDM10(P<0.05).Conclusion5-Aza-CdR could inhibit the cell proliferation of Hs578T,which might be related to the demethylationof PRDM10gene in the cells.
关 键 词:乳腺肿瘤 甲基化 细胞增殖 PRDM10 5-氮杂-2′-脱氧胞苷
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