Mammaglobin Expression And Its Clinical Significance

The concept of micrometastasis is hard to find transfers of non-hematologicaltumor cells by conventional methods,including the diameter of less than 2mmmicro-metastases and beyond the primary tumor tissues and organs in a single livingcell cycle or cell clusters. Most scholars agree with diameter less then 2mm as thecritical standards to judge micrometastases。Studies have confirmed that breast canceris a systemic disease, early breast cancer may have already existed blood,lymph nodeand bone marrow micrometastases。These mico-metastasis in breast cancer patients islikely to be after the recurrence and metastasis of latent lesions。So how do we detectthese subclinical micrometastasis has become essential。Detection of micrometastasis,the key is to find a specific tumor marker。Currently used for detection ofmicrometastases of breast cancer markers are mainly:CK,CEA,CA125,althoughthese indicators to monitor relapse has significance,but because of low specificity,the results are not satisfactory。If you can find a specific,sensitive markers to detect asubclinical micro-metastasis,for breast cancer patient prognosis and guidetreatment,the survival rate of increase and so have important clinical significance。Mammaglobin(or a human mammaglobin,hMAM)has been found in recent years tobe a tissue specific protein that is related with breast cell growth,molecμlar weight is10.5kd。Many studies confirm that mammaglobin only in normal breast tissue,ishighly expressed in breast cancer。But hMAM in normal tissue function and thepathogenesis of breast cancer the role is not quite clear。Geven its high specificity,are more and more widely used as a detection of breast cancer lymph node,bonemarrow and peripheral blood markers of micrometastasis,is a usefμl clinical markerfor the futhre。The current method of detecting breast cancer micrometastases mainly haveimmunohistochemistry and flow cytometry,however,the shortcoming of thesemethods have false positive rate,less sensitive。In recent years,RQ-PCR has beendeveloped,which is based on the general PCR。With its sensitive,rapid,specificand has been widey used。It also reduces the repeated operation of the product to becontamination。And as RQ-PCR technique using specific fluorescent probes,significantly improved the accuracy of quantitative PCR and product detection sensitivity can reach 106-107。Purpose:The establishment of RQ-PCR to detect breast cancer,axillary lymphand peripheral blood hMAM mRNA expression,explore the relationship betweenmammaglobin expression with lymph node,peripheral blood micrometastasis andprognosis,for the clinical exploration of new and valuable prognostic indicator。Method: (1) Conventional methods for extracting tissue and peripheral bloodmRNA reverse transcriptase cDNA for backup。Primers and probes were designedusing Beacon Designer 7.0 design software。Firstly conduct breast cancer hMAMtarget gene and gene GADPH by qualitative PCR,the positive samples wereamplified after extracted of plasmid concentration was measured。And then measureseveral time to copies as the dilution to 107,106,105,104,103 different gradients,making standard curve according to the results by fluorescence quantitative PCR.(2) Samples were collected and extracted mRNA,and RT for the cDNA,Usingsteward genes as the reference,based on standard curve,it measures differentsamples of target gene expression。Resμlt:1. Breast cancer tissue expression of hMAM mRNA:RQ-PCR detdcted15cases of carcinoma of mammglobin expression,accounted for 14cases of positive。Benign tumor in the breast and other cancer tissues were not detected in theexpression of mammaglobin。2．lymph node expression of hMAM mRNA:RQ-PCR detected 30 cases withbreast cancer axillary lymph nodes in the expression of mammaglobin。18cases ofpathologically confirmed lymph node mestastasis detected of which 17 cases ofpositive expression of mammaglobin。Pathological negative lymph nodes,theexpression of hMAM 25%。3．peripheral blood expression of hMAM mRNA:54 cases of breast cancer inperipheral blood hMAM mRNA positive expression rate of 29%。Ⅰ10%,Ⅱ21%,Ⅲ45%,Ⅳ80%。20 cases of breast fibromatosis and 20 cases of healthy volunteerswere not detected in peripheral blood hMAM mRNA expression。The expression ofmammaglobin positive relationship with the clinical stage。In this study,RQ-PCR technique to detect expression of hMAM mRNA in breastcancer,axillary lymph node,peripheral,confirmed that mammaglobin expression in breast cancer tissues,no expression in other tissues,is a highly specific marker forbreast cancer。It may also reflect micrometastasis of breast cancer in the lymphnodes,peripheral blood。It has important clinical signigicance for prognosis andsurvival rate of patients。Conclusion:Given the high specificity of mammaglobin,it wants to be amarker in breast cancer for detection micrometastases。The recent development ofRQ-PCR technology for detection of micrometastases has provided a more accurate,sensitive and convenient method。Micrometastases of breast cancer may beindependent prognostic indicators of breast cancer。...