The Preliminary Analysis Of Intraoperative Diagnosis Method Of Sentinel Lympha Node Metastasis Of Breast Cancer

Immunomagnetic bead separation is a relative new technology. In order to improve from to the deficiency of the conventional detection methods (such as:RT-PCR, flow-cytometry), these methods are often combinded with immunomagnetic bead technology, owing to its high specificity, high isolation rate and without affecting the activities and characteristics of the isolated cells. The value of immunomagnetic bead sepration in detection of circulating tumor cells has been confirmed,but its application in the intraoperative diagonosis of breast cancer sentinel lymph node biopsy (SLNB) technique has not been reported yet.Our research applys immunomagnetic beads technique in sentinel lymph node biopsy introperative diagnosis in early breast cancer patients.We compared the sensitivity, specificity, false-negative rate and other indicators with the frozen HE staining and cytokeratin immunohistochemistry (CK-IHC) method. We attempt to establish a new intraoperative molecular diagnostic approach for the current sentinel lymph node biopsy in breast cancer.Between January and September of 2009, we included 62 consecutive cases of female breast cancer patients in our research. All the patients received sentinel lymph node biopsy surgery. After being labeled, all steniel lymph nodes (SLN) were devided into four parts. The A and D parts were analyzed the expression of lymph nodes metastasis marker(CD44v6,her-2,Mucin-1) through immunomagnetic bead separation and flow-cytometry, we then examined if the lymph nodes had any metastasis.In the mean-time, the B and D parts of the same lymph node were cut 2 slice for the frozen HE staining and anti-CK7 and anti-CKAE1/AE3 cytokeratin immunohistochemistry (CK-IHC) to diagnose of SLN for metastasis. We finally evaluate the the sensitivity, specificity and false negative rate of the 3 different methods.1. Sentinel node biopsy:In the 62 patients, we have successfully identified 52 cases of SLN, the success rate is 83.9% and 89 SLN were identified. All the 52 patients who underwent successful SLNB received full axillary lymph node dissection,914 axillary non-SLN were detected.25 patients with a total of 120 axillary lymph nodes had metastasis.2. Sentinel lymph node HE staining:17 cases of 24 patients were found of SLN metastasis. In the 25 patients who had axillary lymph node metastasis,8 cases did not found lymph node metastasis in frozen section, the false negative rate was 32% (8/ 25). In SLN intraoperative frozen HE staining,the sensitivity was 68%(17/25), and the specificity was 100%(17/17).3. Anti-cytokeratin immunohistochemistry:28 of 19 patients diagnosed with SLN metastasis. Using cytokeratin immunohistochemical staining for diagnosis of sentinel lymph node, the sensitivity was 76.0%(19/25), the specificity was 100.0%(19/19), and the false negative rate was 24% (6/25).4. Immunomagnetic beads sepration:Immunomagnetic separation combined with flow cytometry analysis, in the 62'patients included, we found 24 patients with SLN metastasis. The false negative rate was 4.0%(1/25). the sensitivity was 96% (24/25), the specificity was 96.3% (26/27).1. We apply immunomagnetic separation technology in the intraoperative diagnosis of sentinel lymph node metastasis in the SLNB of breast cancer patients, and compare the sensitivity,speciality and false negativity rate with the traditional frozen HE staining and CK-IHC.We found that immunomagnetic bead separation could detect the magligent cells in the steniel lymph nodes with rapid speed and high accuracy.2. Traditional frozen HE staining was widely used in the intraoperative diagnosis of the stentinel lymph nodes in the breast cancer.But we confirm that traditional frozen HE stating had the dificiency in term of sensitivity and high false negative rate.3. In compared to frozen HE staining,the CK-IHC has distinct advantages in the sensitivity and false negative rate. In combination of frozen HE staining and CK-IHC,we can enhance the sensitivity efficiently.However, the CK-IHC may spent one or two days,which made it less practical in the introperative diagnosis in the SLNB.4. When compared with frozen HE staining and CK-IHC, MACS has distinct advantages in terms of high sensitivity and low false negative rate. Large cohort study are needed to veify our results. 5. The expression of CD44v6,Her-2 and Mucin-1 in the sentinel lyphma nodes were in positivly correlated with the expression of the primary tumor. It is hopeful that the three kinds of biomarkers above can predict whether the SLN has metastasis or not.