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目的探讨抑癌基因TIMP3失活与乳腺癌发生和进展的关系。方法用甲基化特异性PCR技术和亚硫酸盐测序技术检测乳腺癌发生模型MCF10的增生细胞系MCF10A、癌前细胞系MCF10AT、导管内癌细胞系MCF10DCIS.com、浸润癌细胞系MCF10CA1a和转移癌细胞系MCF10CA1d、MCF10CA1h中TIMP3启动子区甲基化状态。结果甲基化特异性PCR分析显示,在上述各细胞系中,TIMP3启动子区均呈高度甲基化状态。亚硫酸盐测序显示,在上述各细胞系中,测序区内的68个CG位点几乎全部发生了甲基化,且甲基化累及了绝大部分等位基因。结论TIMP3基因启动子区甲基化在乳腺癌的发生和进展中起重要作用,可能成为早期诊断乳腺癌和判断乳腺癌预后的分子生物学标记。
Objective To investigate the relationship between inactivation of tumor suppressor gene TIMP3 and the occurrence and progression of breast cancer. Methods MCF10A, MCF10AT, MCF10DCIS, MCF10CA1a and MCF10CA1a were detected by methylation-specific PCR and sulfite sequencing. Cell line MCF10CA1d, MCF10CA1h TIMP3 promoter methylation status. Results Methylation-specific PCR analysis showed that the promoter region of TIMP3 was highly methylated in all the above cell lines. Sulfite sequencing showed that almost all of the 68 CG sites within the sequencing region were methylated in all of the above cell lines and that methylation involved most of the alleles. Conclusions Promoter methylation of TIMP3 gene plays an important role in the occurrence and progression of breast cancer and may be a molecular marker for early diagnosis of breast cancer and prognosis of breast cancer.