人乳房珠蛋白外周血表达RT-PCR检测及乳腺癌诊治应用  

The Detection of Human Mammaglobin(hMAM)Expression in the Peripheral Blood by Reverse Transcriptase Polymerase Chain Reaction Assay and its Application in the Diagnosis of Breast Cancer

作　　者：董学君[1] 黄黎明[1] 施长根 郑专[1] 赵雅清[1] 陈建军[1] 王贵珍[1] 

机构地区：[1]浙江省绍兴市人民医院,312000 [2]上海申友生物技术公司

出　　处：《医学研究通讯》2005年第10期12-14,共3页Bulletin of Medical Research

基　　金：浙江省医学科学研究基金项目(2004A086)

摘　　要：目的建立灵敏外周血人乳房珠蛋白(hMAM)表达 RT-PCR 检测方法,探索 hMAM 在乳腺癌诊治中的应用价值。方法分离受检者外周血单个核细胞,TRIZOL 液提取总 RNA,作 RT-PCR,并查β-actin 监控;确认的 cDNA 先作普通PCR 扩增,再作荧光定量 PCR 扩增。结果 64例标本有63例确认转录为 cDNA,其中乳腺癌43例、良性乳腺肿瘤和正常对照各10例。经普通 PCR 后再作荧光定量 PCR,乳腺癌 hMAM 表达阳性率为53.3%,与良性乳腺肿瘤和正常对照间阳性率有显著差异(P<0.05);而不作普通 PCR 的 cDNA 直接行荧光定量 PCR,结果无阳性。淋巴结转移较未转移,hMAM 表达阳性率有极显著差异(P<0.01)。结论外周血 hMAM 表达是乳腺癌细胞微转移的特异指标;单次 PCR 法的灵敏度不够,二次 PCR 可明显提高灵敏度,并一次普通 PCR 加一次荧光定量 PCR 的组合效果更理想。外周血 hMAM 表达可能是监测淋巴结转移较好指标。Objective To explore the application of hMAM detection in the diagnosis of breast cancer, a sensitive hMAM RT- PCR assay for peripheral blood was tested and well estabhshed. Methods Mononuclear cells from patients＇ peripheral blood were isolated. Total RNA was extract by using TRIZOL. RT PCR was then performed. β- actin gene amplification was used as control. Both PCR and Real - time PCR were apphed after cDNA amplification was achieved in this study. Results 63 of 64 samples had cDNA amplification. Among them, 43 cases were breast cancer, and 10 of them were breast benign lesions and normal controls. By using real time PCR as second round amplification in a nested PCR, hMAM expression was detected in 53.3% of breast cancer patients tested, showing statistic difference （P 〈 0.05） as compared with the breast benign lesions and normal controls. No any positive results were found when only single round real time PCR was applied. Higher hMAM expression was detected in patient group with lympho- nude metastasis, showing statistic different （P 〈 0.01） as compared with patients without lympho- nude metastasis.Conclusion Human mammaglobin （hMAM） expression in the peripheral blood can be an indicator of the metastasis of breast cancer cells. Nested PCR with combination of 1st normal PCR and 2nd real time PCR has improved sensitivity in hMAM gene amplification. The detection of hMAM expression in peripheral blood could also indicate the emergence of lympho - nude metastasis.

关 键 词：乳腺癌 人乳房珠蛋白 RT-PCR 外周血 

分 类 号：R655.8[医药卫生—外科学] R737.9[医药卫生—临床医学]