The Study Of The Quantitative Detection Of HMAM MRNA In Peripheral Blood Of Breast Cancer Patients And Its Clinical Correlations

ObjectiveTo establish SYBR Green I Q-RT-PCR method for detection of the quantification of hMAM mRNA in peripheral blood and breast cancer tissues and corresponding adjacent breast tissues of breast cancer patients;To investigate the feasibility of hMAM mRNA as a marker of CTCs and to analyze the correlations bettween CTCs in the peripheral blood of breast cancer patients and patients' clinical and pathological features.MethodsSYBR Green I Q-RT-PCR was applied to the quantification detection of hMAM mRNA in breast cancer cell line(MDA-MB-231) and peripheral blood samples from 61 patients with breast cancer and 33 patients with benign breast tumor and 11 healthy volunteers. Furthermore,10 breast cancer tissues and corresponding adjacent breast tissues were also detected.Results1.SYBR Green I Q-RT-PCR could detect hMAM mRNA expression with high CT value of 34.45~34.67(CV=0.25%) in breast cancer cell line(MDA-MB-231) repeatedly,hMAM mRNA was negative in peripheral blood of the benign breast tumor patients and healthy volunteers.Therefore this method had a good sensitivity,reproducibility and specificity.2.hMAM mRNA was expressed in 10/61(16.4%) peripheral blood of breast cancer patients:0/3 in stages 0, 0/5 in stagesⅠ, 5/33(15.15%)in stagesⅡand 5/20(25.0%)in stagesⅢ. 3/31(9.7%)in No, 2/12(6.7%)in N1, 4/15(26.7%)in N2 and 1/3(33.3%)in N3. hMAM mRNA-positive ratio increased with the increasing of the disease stages and node status,but the differences had not statistical significance (P>0.05).10 cases hMAM mRNA-positive in peripheral blood of breast cancer patients, 8 cases were HER2 overexpression in primary tumors, no cases were HER2-negative. The difference had significant statistically(P=0.028). The relative quantification of hMAM mRNA in peripheral blood of breast cancer patients were in range of 1.98×10~2～6.21×10~3, (X|-)±S=1119.9±1829.4(median 4.0×10~2). There were not significant difference between the relative quantification of hMAM mRNA in peripheral blood of breast cancer patients and patients' clinical and pathological features(P>0.05).3.hMAM mRNA was detected in 8/10(80%) breast cancer tissues: 1/1(100%) in stagesⅠ, and the relative quantification was 37800; 6/6(100%)in stagesⅡ, and the median relative quantification was 100.4; 1/3(33.3%)in stagesⅢ, the relative quantification was 2.94. Two patients were hMAM mRNA-negative in both of their cancer tissues and corresponding adjacent breast tissues. Breast cancer tissues and their corresponding adjacent breast tissues co-expressed hMAM mRNA.The quantification of hMAM mRNA expression in breast cancer tissues had no significant difference correlation to patients'clinical and pathological features because of the small cases.Conclusions1.SYBR Green I Q-RT-PCR method can be used to detect the quantification of CTCs in peripheral blood of breast cancer patients and also can be used to detect the quantification of hMAM mRNA in breast tissues.2.There was a certain proportion of hMAM mRNA expression in peripheral blood of breast cancer patients, and significantly related to the HER2 status of the primary tumor. Breast cancer tissues and corresponding adjacent breast tissues had high proportion and high levels of hMAM mRNA expression,few of breast cancer patients had no or low hMAM mRNA expression in their cancer tissues and corresponding adjacent breast tissues.3.The detection of hMAM mRNA in peripheral blood of breast cancer may be used as a indicator of CTCs.4.The significance of the quantification of CTCs in peripheral blood of breast cancer patients should be analyzed with the results of following-up.