摘要
目的:研究5-氮杂2′-脱氧胞苷(5-Aza-CdR)对MCF-7乳腺癌细胞株生长周期及凋亡的影响,探讨其临床治疗乳腺癌的可能性。方法:分别使用浓度为0.4、1.6、6.4、25.6和102.4μmol/L的甲基化抑制剂5-Aza-CdR处理MCF-7乳腺癌细胞株。通过MTT检测5-Aza-CdR对MCF-7乳腺癌细胞株存活率的影响。应用流式细胞仪检测5-Aza-CdR对MCF-7乳腺癌细胞株生长周期及凋亡的影响。RT-PCR检测处理前后抑癌基因RASSF1A mRNA在MCF-7乳腺癌细胞株表达的变化。结果:5-Aza-CdR6.4μmol/L作用1d或0.4μmol/L作用3d就可以显著抑制MCF-7乳腺癌细胞的增殖,P<0.05。作用3d后,在6.4μmol/L时细胞周期中处于G0/G1期的细胞的显著增加(P<0.05),细胞周期阻滞于G0/G1期;在0.4μmol/L时细胞的凋亡率为(11.80±0.30)%,与对照组比较有显著升高,P<0.01。以上作用呈时间-剂量依赖性关系。5-Aza-CdR处理后,无RASSF1A表达的MCF-7乳腺癌细胞株可检测出基因RASSF1A的重新表达。结论:5-Aza-CdR可消除RASSF1A启动子甲基化状态,使其重新表达而抑制MCF-7乳腺癌细胞株的生长,使细胞周期阻滞于G0/G1期,并促进其凋亡。
OBJECTIVE：To explore the effect of methylation inhibitor 5-Aza-2′-deoxycytidine （5-Aza-CdR） on the growth of human breast cancer cell line MCF-7, and to inves- tigate the possibility of its application in clinical treatment. METHODS： Human breast cancer MCF-7 cells were treated with 0.4, 1.6, 6.4, 25.6 and 102.4 μmol/L 5-Aza-CdR, respectively. Then the growth of the cells were observed by MTT assay. The cell cycle and apoptosis were analyzed by flow cytometry. The expression of the tumor suppressor gene RASSF1AmRNA was detected by reverse transcription polymerase chain reaction （RT-PCR）. RESULTS： 5-Aza-CdR inhibited the proliferation of MCF-7 cells significantly at a concentration of 1.6 μmol/L for 1 day or at a concentration of 0.4 μmol/L for 3 days （P〈0.05） ; after treatment for 3 days, the number cells of G0/G1 phase increased at a concentration of 6.4 μmol/L （P〈0.05）, and the cell cycle was blocked at G0/G1 phase. The apoptosis rate increased significantly to 11.8± 0.30% at a concentration of 0.4μmol/L compared with the control group （P〈0.01）. The effects mentioned above were in a time- and concentration-dependent manner. RASSFIA gene was reactivated by 5-Aza-CdR in MCF-7 cells not expressing RASSFIA. CONCLUSION： 5-Aza-CdR can inhibit the growth, and promote the apoptosis of MCF-7 cells by eliminating the methylation status of RASSFIA promoter.
出处
《中华肿瘤防治杂志》
CAS
2008年第6期415-418,共4页
Chinese Journal of Cancer Prevention and Treatment
关键词
脱氧胞苷
乳腺肿瘤
细胞周期
细胞凋亡
deoxycytidine
breast neoplasms
cell cycle
cell apoptosis
