天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (3): 234-238.doi: 10.11958/20170981

• 细胞与分子生物学 • 上一篇    下一篇

5-氮杂-2′-脱氧胞苷对人乳腺癌细胞Hs578T增殖和PRDM10基因甲基化的影响

张晓宇，白杰，康晓宁，靳丽君，王鹏，刘伟，王遵义   

  1. 1河北省沧州市中心医院肿瘤外三科（邮编061001），2超声二科
  • 收稿日期:2017-09-19 修回日期:2018-01-05 出版日期:2018-03-15 发布日期:2018-03-23
  • 通讯作者: 张晓宇 E-mail:93956466@qq.com

Effects of 5- Aza -2′- deoxycytidine on proliferation of human breast cancer cell line Hs578T and methylation of PRDM10 gene

ZHANG Xiao-yu,BAI Jie,KANG Xiao-ning,JIN Li-jun,WANG Peng,LIU Wei,WANG Zun-yi   

  1. 1 Third Department of Oncological Surgery, 2 Second Department of Ultrasonography, Cangzhou Central Hospital, Heibei 061001, China
  • Received:2017-09-19 Revised:2018-01-05 Published:2018-03-15 Online:2018-03-23

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摘要： 目的 分析 5-氮杂-2′-脱氧胞苷（5-Aza-CdR）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、5 μmol/L 的 5-Aza-CdR 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 mRNA 和蛋白表达水平。结果 MTT 结果显示，5-Aza-CdR 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 0 μmol/L 组比较，1 μmol/L 组处理 72 h 后及 3、5 μmol/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-CdR 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-CdR 的浓度越高，PRDM10 的 mRNA和蛋白表达水平越高（P＜0.05）。结论 5-Aza-CdR 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。

关键词: 乳腺肿瘤, 甲基化, 细胞增殖, PRDM10, 5-氮杂-2′-脱氧胞苷

Abstract: Abstract: Objective To investigate effects of 5-Aza-2′ - deoxycytidine (5-Aza-CdR) on proliferation of human breast cancer cell line Hs578T, and the methylation status of PRDM10 gene in vitro in this cell line. Methods The human breast cancer cell line Hs578T was cultured with 1, 3 and 5 μmol/L DNA methylation inhibitor 5-Aza-CdR respectively,and untreated cells were used as control. Cell proliferation was detected by MTT assay. Methylation-Specific PCR (MSP) was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by RT-PCR and Western blot assay. Results MTT results showed that the higher the concentration of 5-Aza-CdR, and the longer the treatment time, the more significant inhibitory effect on the proliferation of Hs578T cells. Compared with the control group (0 μmol/L), the proliferation of Hs578T was significantly inhibited after the treatment for 72 h in the 1 μmol/L group, and for 48 h in the 3 μmol/L and 5 μmol/L groups (P＜0.05). MSP results showed that the higher the concentration of 5-Aza-CdR, the more significant demethylation of PRDM10. Results of RT-PCR and Western blot showed that the higher the concentration of 5-Aza-CdR, the higher the expression levels of mRNA and protein in PRDM10 (P＜0.05). Conclusion 5-Aza-CdR could inhibit the cell proliferation of Hs578T, which might be related to the demethylation of PRDM10 gene in the cells.

Key words: breast neoplasms, methylation, cell proliferation, PRDM10, 5-Aza-2′-deoxycytidine

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