【摘要】 目的:检测乳腺癌患者肿瘤循环DNA的含量及其p16基因5′CpG岛甲基化状态的改变.方法:收集61例乳腺癌患者、33例乳腺良性病变患者和27例健康志愿者的血浆样本,抽提血浆循环DNA,以SYBRgreenI荧光染色法行DNA定量;利用半巢式甲基化特异性PCR技术,检测61例乳腺癌患者血浆循环DNA和相应癌组织p16基因5′CpG岛甲基化状态的改变.结果:乳腺癌患者、乳腺良性病变患者、健康自愿者肿瘤循环DNA浓度分别为(65.0±45.3),(19.0±9.5)和(13.0±7.3)μg/L,乳腺癌患者血浆循环DNA浓度显著高于乳腺良性病变患者和健康自愿者,差异有统计学意义(P<0.05).61例乳腺癌患者血浆循环DNA和相应癌组织p16基因5′CpG岛甲基化状态的改变检出率分别为44.3%和46.2%,两者检出率无统计学差别(P>0.05).结论:血浆肿瘤循环DNA的定量有可能成为一种新的恶性肿瘤标志物.更多还原

【Abstract】 AIM:To quantify the circulating DNA and detect the variation of 5’ CpG hypermethylation in p16 gene in plasma from patients with breast cancer.METHODS:Blood samples were collected from 61 breast cancer patients,and 33 patients with non-neoplastic breast diseases,and 27 healthy volunteers.The circulating DNA was extracted and analyzed by fluorescent dye(SYBR green I)staining.Using a modified semi-nested methylation-specific PCR(MSP),the status of methylation of p16 gene was detected in circulating plasma DNA and corresponding breast cancer tissues.RESULTS:The circulating DNA concentration in the breast cancer was(65.0±45.3)μg/L,significantly higher than that in the patients with non-neoplastic breast diseases(19.0±9.5)μg/L and that in the normal controls [(13.0±7.3)μg/L,P<0.05].Methylated p16 gene was found in 44.3%(27/61)circulating DNA and in 46.2% tissues of breast cancer,and there was no significant difference between them(P>0.05).CONCLUSION:Circulating DNA in plasma may become an additional tumor marker for the diagnosis of breast cancer.更多还原