抑制DBC2基因启动子甲基化促进乳腺癌细胞凋亡的研究

发布时间：2018-08-08 15:21

【摘要】：目的研究抑癌基因DBC2在乳腺癌的表达与甲基化状态之间的联系,及其对乳腺癌细胞系T-47D细胞增殖和凋亡的影响。方法体外培养3种乳腺癌细胞株(T-47D、MCF-7、MDA-MB-23)和正常乳腺细胞株(MCF-10),检测4种细胞中DBC2基因的表达及其启动子区甲基化状况;5-Aza-Cd R处理T-47D细胞后,检测5-Aza-Cd R对T-47D细胞DBC2启动子甲基化状态和其表达的影响,并通过MTT和Annexin-V/PI双染法分别检测处理前后T-47D细胞增殖和凋亡的变化。结果 DBC2在T-47D细胞表达缺失,乳腺癌细胞DBC2启动子发生甲基化;5-Aza-Cd R成功逆转了T-47D细胞DBC2的甲基化状态,并使DBC2 m RNA表达恢复(P0.05);不同浓度的5-Aza-Cd R处理T-47D细胞后,T-47D细胞增殖下降(P0.05),凋亡率增加(P0.05)。结论 DBC2启动子甲基化导致T-47D细胞DBC2表达沉默,激活DBC2表达可抑制T-47D细胞增殖并促进其凋亡。
[Abstract]:Objective to investigate the relationship between the expression of tumor suppressor gene DBC2 and methylation status in breast cancer and its effect on the proliferation and apoptosis of breast cancer cell line T-47D. Methods three kinds of breast cancer cell lines (T-47D- MCF-7MDA-MB-23) and normal mammary gland cell line (MCF-10) were cultured in vitro. The expression of DBC2 gene and the methylation status of its promoter region were detected after T-47D cells were treated with 5-Aza-CD R. The effects of 5-Aza-Cd R on the methylation status and expression of DBC2 promoter in T-47D cells were detected. The proliferation and apoptosis of T-47D cells were detected by MTT and Annexin-V/PI double staining. Results the expression of DBC2 was absent in T-47D cells, and the methylation of DBC2 promoter in breast cancer cells was successfully reversed by 5-Aza-CD R, which reversed the methylation of DBC2 in T-47D cells. The expression of DBC2 m RNA was restored (P0.05), and the proliferation of T-47D cells decreased (P0.05) and the apoptosis rate increased (P0.05) after treated with different concentrations of 5-Aza-Cd R (P0.05). Conclusion methylation of DBC2 promoter induces silencing of DBC2 expression in T-47D cells, and activation of DBC2 expression can inhibit the proliferation of T-47D cells and promote their apoptosis.
【作者单位】：河南省中医院乳腺科;
【分类号】：R737.9

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