目的建立1种简便、有效的SYBR Green Ⅰ实时定量逆转录PCR(SYBR Green Ⅰ Q—RT—PCR)方法检测乳腺癌组织、癌旁乳腺组织人乳腺珠蛋白(hMAM)mRNA表达水平。方法对SYBR Green Ⅰ Q—RT—PCR检测乳腺癌细胞株MDA—MB—231表达hMAM mRN...目的建立1种简便、有效的SYBR Green Ⅰ实时定量逆转录PCR(SYBR Green Ⅰ Q—RT—PCR)方法检测乳腺癌组织、癌旁乳腺组织人乳腺珠蛋白(hMAM)mRNA表达水平。方法对SYBR Green Ⅰ Q—RT—PCR检测乳腺癌细胞株MDA—MB—231表达hMAM mRNA的反应条件进行优化,初步检验该方法的重复性、灵敏性。应用该方法检测11例健康志愿者外周血hMAM mRNA表达,检验该方法的特异性,并对10例乳腺癌组织、10例对应癌旁乳腺组织hMAM mRNA表达水平进行测定。结果SYBR Green Ⅰ Q—RT—PCR可重复栓出乳腺癌细胞株MDA—MB-231极低表达hMAM mRNA,CT值为34.45~34.67,变异系数(CV)=0.25%。健康志愿者外周血中hMAM mRNA均阴性。乳腺癌组织hMAM mRNA阳性率为80.0%(8/10),其中Ⅰ期1例(100.0%),其量为37800;Ⅱ期6例(100.0%),量为100.4(中位);Ⅲ期1例(33.3%),量为2.94。结论建立的SYBR Green Ⅰ Q—RT—PCR方法具有简便、特异、重复性、灵敏度好、定量结果可靠、直观等优点,应用于乳腺癌组织和癌旁乳腺组织hMAM mRNA的定量检测是理想的。展开更多
目的:建立SYBR Green I实时定量逆转录PCR(Real-Ti me QuntitativeRT-PCR,Q-RT-PCR)技术定量检测人乳腺珠蛋白(human mammaglobin,hMAM)mRNA的技术,并应用于检测外周血hMAM mRNA,探讨其诊断乳腺癌患者外周血微转移的可行性及意义...目的:建立SYBR Green I实时定量逆转录PCR(Real-Ti me QuntitativeRT-PCR,Q-RT-PCR)技术定量检测人乳腺珠蛋白(human mammaglobin,hMAM)mRNA的技术,并应用于检测外周血hMAM mRNA,探讨其诊断乳腺癌患者外周血微转移的可行性及意义。方法:应用自行建立的SYBR Green I Q-RT-PCR检测技术,检测61例乳腺癌、33例乳腺良性肿瘤患者以及11例健康志愿者外周血hMAM mRNA的表达量,分析其表达水平与乳腺癌临床病理特征及ER、PR及HER-2表达的关系。结果:61例乳腺癌患者外周血中10例检出hMAM mRNA表达,其癌组织HER-2阳性者占8例,HER-2阴性组未见hMAM mRNA阳性表达。HER-2阳、阴性组间外周血hMAM mRNA表达,差异具有统计学意义,P=0.028;与其他临床、病理特征无相关性,P〉0.05。HER-2阳性患者外周血中hMAM mRNA表达量范围1.98×10^2-6.21×1^03,中位数4.0×10^2,在临床、病理特征间差异无统计学意义,P〉0.05。乳腺良性肿瘤及健康志愿者外周血hMAM mRNA均阴性。结论:SYBR GreenI Q-RT-PCR技术可定量检出乳腺癌患者外周血hMAM mRNA;乳腺癌患者外周血hMAM mRNA表达与其乳腺癌组织HER-2表达相关。定量检测hMAM mRNA可作为乳腺癌外周血微转移的重要方法。展开更多
Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Methods: S...Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Methods: SBEM and hMAM expressions in PB samples of 109 primary breast cancer patients were detected by flow cytometry (FCM) and RT-PCR. Relationship between the biomarkers' expression and prognostic parameters were analyzed. Results: SBEM and hMAM expressions in PB of breast cancer patients were much higher than those of healthy donors and other cancer patients. SBEM and hMAM expressed in 53.2% (50/94) and 39.4% (37/94) cases at stages Ⅰ-Ⅲ and expressed in 73.3% (11/15) and 46.7% (7/15) cases at stage IV respectively. SBEM and hMAM mRNA were only detected in PB samples of breast cancer patients, while no expression of them was found in that of healthy donors and other cancer patients. Conclusion: hMAM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-positive, well-differentiated breast cancers and SBEM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-negative, poorly-differentiated breast cancers.展开更多
文摘目的建立1种简便、有效的SYBR Green Ⅰ实时定量逆转录PCR(SYBR Green Ⅰ Q—RT—PCR)方法检测乳腺癌组织、癌旁乳腺组织人乳腺珠蛋白(hMAM)mRNA表达水平。方法对SYBR Green Ⅰ Q—RT—PCR检测乳腺癌细胞株MDA—MB—231表达hMAM mRNA的反应条件进行优化,初步检验该方法的重复性、灵敏性。应用该方法检测11例健康志愿者外周血hMAM mRNA表达,检验该方法的特异性,并对10例乳腺癌组织、10例对应癌旁乳腺组织hMAM mRNA表达水平进行测定。结果SYBR Green Ⅰ Q—RT—PCR可重复栓出乳腺癌细胞株MDA—MB-231极低表达hMAM mRNA,CT值为34.45~34.67,变异系数(CV)=0.25%。健康志愿者外周血中hMAM mRNA均阴性。乳腺癌组织hMAM mRNA阳性率为80.0%(8/10),其中Ⅰ期1例(100.0%),其量为37800;Ⅱ期6例(100.0%),量为100.4(中位);Ⅲ期1例(33.3%),量为2.94。结论建立的SYBR Green Ⅰ Q—RT—PCR方法具有简便、特异、重复性、灵敏度好、定量结果可靠、直观等优点,应用于乳腺癌组织和癌旁乳腺组织hMAM mRNA的定量检测是理想的。
文摘Objective: The aim of the study was to explore the individual detection significances of small breast epithelial mucin (SBEM) and human mammaglobin (hMAM) in peripheral blood (PB) of breast cancer patients. Methods: SBEM and hMAM expressions in PB samples of 109 primary breast cancer patients were detected by flow cytometry (FCM) and RT-PCR. Relationship between the biomarkers' expression and prognostic parameters were analyzed. Results: SBEM and hMAM expressions in PB of breast cancer patients were much higher than those of healthy donors and other cancer patients. SBEM and hMAM expressed in 53.2% (50/94) and 39.4% (37/94) cases at stages Ⅰ-Ⅲ and expressed in 73.3% (11/15) and 46.7% (7/15) cases at stage IV respectively. SBEM and hMAM mRNA were only detected in PB samples of breast cancer patients, while no expression of them was found in that of healthy donors and other cancer patients. Conclusion: hMAM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-positive, well-differentiated breast cancers and SBEM mRNA detection maybe helpful to predict hematogenous micrometastasis in ER-negative, poorly-differentiated breast cancers.