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目的明确表观遗传学药物对乳腺癌细胞系抑癌基因BRCA1及CHD5表达的作用,探讨此类药物在乳腺癌临床治疗中的合理应用。方法采用去甲基化制剂5-AZA和组蛋白去乙酰化酶抑制剂TSA分别作用于乳腺癌细胞系T47D、MCF-7及乳腺正常上皮细胞HBL-100,5-AZA终浓度分别为0、0.5、1.0、2.5、5.0和10.0μmol/L,TSA终浓度分别为0、0.25、0.5、0.75、1.0和1.5μmol/L。荧光定量RT-PCR检测各浓度5-AZA作用12、24、48及72 h,各浓度TSA作用6、12、24、48及72 h BRCA1和CHD5的表达水平,并行甲基化特异PCR(MSP)检测两基因的甲基化状态。结果 2.5μmol/L AZA和0.5μmol/L TSA作用可提高乳腺癌细胞系BRCA1及CHD5的表达,而10μmol/L AZA和1.0、1.5μmol/L TSA作用反致表达降低,且对乳腺正常上皮细胞呈毒性作用。MSP未能发现AZA和TSA对MCF-7细胞两抑癌基因及T47D细胞BRCA1甲基化状态的影响,而两种药物均可逆转T47D细胞CHD5的甲基化状态。结论表观遗传学药物对乳腺癌细胞抑癌基因表达的影响具有给药剂量及时间限制性,本研究可为此类药物在乳腺癌治疗中的科学应用提供实验依据。
Objective To clarify the role of epigenetic drugs in the expression of tumor suppressor genes BRCA1 and CHD5 in breast cancer cell lines and to explore the rational application of these drugs in the clinical treatment of breast cancer. Methods The final concentration of 5-AZA and histone deacetylase inhibitor TSA in breast cancer cell lines T47D, MCF-7 and normal breast epithelial cells HBL-100,5-AZA were 0, 0.5, 1.0, 2.5, 5.0 and 10.0 μmol / L with final TSA concentrations of 0, 0.25, 0.5, 0.75, 1.0 and 1.5 μmol / L, respectively. The expression of BRCA1 and CHD5 were detected by real-time RT-PCR at different concentrations of 5-AZA for 12, 24, 48 and 72 h, and at various concentrations of TSA for 6, 12, 24, 48 and 72 h, ) To examine the methylation status of both genes. RESULTS: 2.5μmol / L AZA and 0.5μmol / L TSA could enhance the expression of BRCA1 and CHD5 in breast cancer cell lines, but the expression of BRCA1 and CHD5 was decreased in 10μmol / L AZA and 1.0,1.5μmol / L TSA, Toxic effect. MSP failed to find the effect of AZA and TSA on the methylation status of BRCA1 in both Tumor suppressor gene and T47D cells in MCF-7 cells. However, both drugs reversed the methylation status of CHD5 in T47D cells. Conclusion Epigenetic drugs have a dose-dependent and time-limited effect on tumor suppressor gene expression in breast cancer cells. This study may provide experimental evidence for the scientific application of these drugs in the treatment of breast cancer.