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目的探讨乳腺癌发生模型MCF10中抑癌基因APC启动子区甲基化状态及其对mRNA表达的影响。方法应用甲基化特异性聚合酶链反应(MSP)及双亚硫酸钠基因测序技术检测MCF10模型的乳腺增生细胞系MCF10A、癌前细胞系MCF10AT、导管内癌细胞系MCF10DC IS.com、浸润癌细胞系MCF10CA1 a、MCF10CA1d、MCF10CA1h及经典乳腺癌细胞系MCF-7和正常乳腺组织中APC基因启动子1A甲基化状态,然后用逆转录聚合酶链反应(RT-PCR)和实时PCR技术检测上述样品的mRNA表达水平。结果在MCF10模型的增生细胞系、癌前细胞系、导管内癌细胞系、浸润癌细胞系中,APC基因启动子1A处于低甲基化状态;与正常乳腺组织相比,各细胞系APC基因mRNA表达无明显减少,MCF10AT、MCF10CA1d、MCF10CA1h、MCF10DC IS.com的mRNA表达分别减少0.27、0.96、1.78、2.70、2.03倍,MCF10A和MCF-7分别增加0.02和0.33倍)。结论MCF10模型中乳腺癌的发生发展过程与APC基因启动子区异常甲基化无关。
Objective To investigate the methylation status of tumor suppressor gene APC promoter region and its effect on mRNA expression in MCF10 breast cancer. Methods MCF10 breast cancer cell line MCF10A, MCF10AT, MCF10DC IS.com, invasive cancer cell line MCF10 were detected by methylation-specific polymerase chain reaction (MSP) and bisulfite gene sequencing MCF10CA1 a, MCF10CA1d, MCF10CA1h and APC gene promoter methylation status in the classical breast cancer cell line MCF-7 and normal breast tissue, and then the above samples were detected by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR techniques MRNA expression level. Results APC gene promoter 1A was hypomethylated in MCF10 model hyperplastic, precancerous, ductal and invasive carcinoma cell lines. Compared with normal breast tissue, APC gene The mRNA expression of MCF10AT, MCF10CA1d, MCF10CA1h and MCF10DC IS.com decreased by 0.27, 0.96, 1.78, 2.70 and 2.03 times, respectively. The mRNA expression of MCF10A and MCF-7 increased by 0.02 and 0.33 fold, respectively. Conclusion The occurrence and development of breast cancer in MCF10 model have nothing to do with the abnormal methylation of APC gene promoter region.