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目的探讨乳腺癌发生过程中抑癌基因NOEY2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PCR及双亚硫酸钠基因测序技术检测MCF10模型中乳腺增生细胞系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及正常乳腺组织中NOEY2基因启动子区CpG岛I甲基化状态,然后用RT-PCR和实时PCR技术检测上述样品的mRNA表达水平。结果MCF10模型的增生细胞系、癌前细胞系、导管内癌细胞系、浸润癌细胞系均发生该基因启动子区CpG岛I高度甲基化;与正常乳腺组织相比,上述细胞系mRNA表达显著减少。结论NOEY2基因启动子区高度甲基化及相应的mRNA表达减少是乳腺癌发生过程中的早期事件,与乳腺癌发生有关,可能成为早期诊断乳腺癌的潜在分子生物学标记。
Objective To investigate the methylation status of tumor suppressor gene NOEY2 promoter region in mammary carcinogenesis and its effect on mRNA expression. Methods MCF10A, MCF10AT, MCF10DC IS, MCF10CA1a, MCF10CA1d and MCF10CA1h were detected by methylation-specific PCR and sodium bisulfite sequencing in MCF10 breast cancer cells And normal breast tissue NOEY2 gene promoter CpG island I methylation status, and then RT-PCR and real-time PCR detection of mRNA expression levels of the above samples. Results The hypermethylation of CpG island I in promoter region of MCF10 model hyperplasia cell line, precancerous cell line, intraductal carcinoma cell line and invasive carcinoma cell line was highly methylated. Compared with normal breast tissue, the mRNA expression of these cell lines Significantly reduced. Conclusion The high methylation level of NOEY2 gene promoter and the corresponding reduction of mRNA expression are early events in breast carcinogenesis, which may be a potential molecular biomarker for the early diagnosis of breast cancer.