【摘要】 目的探讨乳腺癌组织14-3-3 sigma 基因甲基化状态和表达情况。方法用甲基化特异性聚合酶链反应(MSP)方法,分别检测40例乳腺癌患者肿瘤组织和18例乳腺良性病变组织的14-3-3 sigma 基因甲基化状态、并用逆转录(RT)-PCR 和免疫印迹法(WB)分别从 mRNA 和蛋白质水平检测14-3-3 sigma 基因的表达情况。结果在40例乳腺癌患者肿瘤组织中,MSP 法检测出34例14-3-3 sigma 基因甲基化,甲基化率达85%;而 RT-PCR 为12.5%(5/40);WB 法显示,80%患者乳腺癌组织存在14-3-3 sigma 蛋白表达缺失(32/40);在34例 MSP 甲基化患者中,有30例 RT-PCR 和 WB同时显示阴性(88%)。18例良性疾病患者病变乳腺组织均未检出14-3-3 sigma 基因甲基化,而 RT-PCR 和 WB 同时也证明14-3-3 sigma 基因在乳腺上皮细胞均有表达。提示乳腺癌时14-3-3 sigma 基因高度甲基化,且可能与14-3-3 sigma 基因表达缺失密切相关。结论 14-3-3 sigma 基因甲基化及其表达缺失是乳腺癌的经常性事件,14-3-3 sigma 基因甲基化可能与其基因表达缺失有关。更多还原

【Abstract】 Objective To observe the methylation and expression of 14-3-3 sigma gene in human breast cancer.Methods 40 breast cancer tissues and 18 mammary gland tissues with benign lesions were analyzed by methylation specific PCR(MSP),RT-PCR,and Western-blot(WB)so as to detect the methylation status and expression of 14-3-3 sigma mRNA or protein.Results Methylation of 14-3-3 sigma gene was detectable in 85%(34/40)of patients with breast cancers.RT-PCR showed negative in 12.5% of breast cancers(5/40),WB also indicated that 14-3-3 sigma was not detected in 32 of 40 breast carcinomas (80%).Furthermore,both RT-PCR and WB were negative in 30 of 34 positive cases by MSP.While methylation of 14-3-3 sigma was not detectable and its expression was demonstrated by RT-PCR and WB among 18 cases of benign breast diseases.These evidences proposed that methylation of 14-3-3 sigma gene had great relevance with its silence.Conclusion Methylation and loss of expression in 14-3-3 sigma gene were high frequent events in breast cancers.And methylation of 14-3-3 sigma gene might be related to its loss.更多还原