Detection of micrometastasis in fixed paraffin-embedded Sentinel Lymph Nodes of Breast cancer using RT-PCR / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy is considered a highly accurate and very economic method of assessing the axillary nodal status in breast cancer patients. Recently immunohistochemical (IHC) staining and reverse transcriptase polymerase chain reaction (RT-PCR) are commonly used to evaluate micrometastasis in the sentinel lymph node. However, most of the RT-PCR studies have been performed using fresh tissue. This study was conducted to assess micrometastasis in clinically node-negative breast cancer by using RT-PCR technique on the paraffin embedded sentinel lymph nodes. METHODS: Sixty patients who undergone SLN biopsy followed by axillary lymph node dissection due to breast carcinoma were evaluated from February 2000 to January 2001 at the Breast Cancer Center, Department of Surgery, Yongdong Severance Hospital. Serial sections were made from all sentinel lymph nodes for the H&E staining and for the IHC staining with monoclonal anti-cytokeratin antibody. RNA was extracted from the paraffin embedded sentinel lymph nodes and RT-PCR was performed using cytokeratin 19 mRNA, MUC-1 mRNA, and MAGE-A3 mRNA. RESULTS: In 32 out of 60 cases, beta-actin mRNA was detected after RT-PCR, and the 28 cases which had no product after RT-PCR for beta-actin were excluded from this study. Twenty five cases showed as being metastasis positive and 7 cases showed as being metastasis negative by serial section (SS) H&E staining. Three out of 25 negative cases tested for by SS H&E staining were found to be positive by IHC. Ten, six and, eight cases out of the 25 negative cases tested for by SS H&E were found to be positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Among the 22 cases that were found to be negative by both SS H&E staining and IHC staining, 9, 4, and 6 cases were converted to positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Using the combination of two or three markers for performing RT-PCR was more sensitive than any single marker to detect micrometastasis (p < 0.05). CONCLUSION: Even though we failed to extract RNA in 46% of the paraffin embedded tissues, it may be possible to detect micrometastasis by using RT-PCR with the paraffin embedded tissue. RT-PCR is far more sensitive than IHC for detecting microme tastasis, and when we combine multiple markers, the detection rate is higher than for any one marker.

Detection of micrometastasis in fixed paraffin-embedded Sentinel Lymph Nodes of Breast cancer using RT-PCR / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy is considered a highly accurate and very economic method of assessing the axillary nodal status in breast cancer patients. Recently immunohistochemical (IHC) staining and reverse transcriptase polymerase chain reaction (RT-PCR) are commonly used to evaluate micrometastasis in the sentinel lymph node. However, most of the RT-PCR studies have been performed using fresh tissue. This study was conducted to assess micrometastasis in clinically node-negative breast cancer by using RT-PCR technique on the paraffin embedded sentinel lymph nodes. METHODS: Sixty patients who undergone SLN biopsy followed by axillary lymph node dissection due to breast carcinoma were evaluated from February 2000 to January 2001 at the Breast Cancer Center, Department of Surgery, Yongdong Severance Hospital. Serial sections were made from all sentinel lymph nodes for the H&E staining and for the IHC staining with monoclonal anti-cytokeratin antibody. RNA was extracted from the paraffin embedded sentinel lymph nodes and RT-PCR was performed using cytokeratin 19 mRNA, MUC-1 mRNA, and MAGE-A3 mRNA. RESULTS: In 32 out of 60 cases, beta-actin mRNA was detected after RT-PCR, and the 28 cases which had no product after RT-PCR for beta-actin were excluded from this study. Twenty five cases showed as being metastasis positive and 7 cases showed as being metastasis negative by serial section (SS) H&E staining. Three out of 25 negative cases tested for by SS H&E staining were found to be positive by IHC. Ten, six and, eight cases out of the 25 negative cases tested for by SS H&E were found to be positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Among the 22 cases that were found to be negative by both SS H&E staining and IHC staining, 9, 4, and 6 cases were converted to positive by RT-PCR for cytokeratin 19, MUC-1, and MAGE-A3, respectively. Using the combination of two or three markers for performing RT-PCR was more sensitive than any single marker to detect micrometastasis (p < 0.05). CONCLUSION: Even though we failed to extract RNA in 46% of the paraffin embedded tissues, it may be possible to detect micrometastasis by using RT-PCR with the paraffin embedded tissue. RT-PCR is far more sensitive than IHC for detecting microme tastasis, and when we combine multiple markers, the detection rate is higher than for any one marker.

Intraoperative Examination of Sentinel Lymph Nodes Using Rapid Cytokeratin Immunohistochemical Staining in Breast Cancer Patients / 한국유방암학회지

PURPOSE: Sentinel lymph node (SLN) biopsy has become a new standard procedure in the treatment of patients with early breast cancer. Furthermore, many institutions have begun offering the sentinel lymph node biopsy without simultaneous axillary dissection as a possible standard procedure when the SLN was free from tumors. For appropriate intraoperative decision making on the presence of cancer cells in axillary lymph nodes, a fast and accurate method to assess the SLN is required. The authors performed a prospective investigation of the relative merits of rapid cytokeratin immunohistochemical (IHC) staining of the SLN removed during the operations of breast cancer patients. METHODS: Between December 2002 and August 2003, 38 patients with T1and T2 breast cancer were enrolled after undergoing successful sentinel lymph node biopsy. A total of 60 sentinel lymph nodes (mean number, 1.58) were biopsied and first examined by hematoxylin-eosin (H&E) staining. All the tumor free sentinel lymph nodes by H&E stained section were immunostained for cytokeratin using a rapid immunohistochemical assay (Cytokeratin (PAN), 1:50, Newcastle, UK) during the operation. Finally, the sentinel lymph nodes were submitted for paraffin embedding and serial section after surgery. Both H&E stained and cytokeratin immunostained sections were also performed. RESULTS: This technique has a turnaround time of less than 20 minutes during the operation. Rapid IHC staining revealed 4 positive sentinel lymph nodes that were negative for metastasis by H&E staining. Among these false negative 4 cases, two cases had problems with the frozen section of H&E staining and the other 2 cases had problems due to micrometastasis. This study showed a sensitivity of 88.89%, a specificity of 100%, an accuracy of 98.33%, and a negative predictive value of 98.08%. The false-negative case (1 of 52), which was negative on H&E staining and rapid IHC staining during the operation, was disclosed as positive only after a serial permanent section examination with IHC stain. CONCLUSION: The introperative examination of sentinel lymph nodes is a highly accurate and effective way of predicting the axillary lymph node status of patients with breast cancers. This may be a promising technique in deciding whether to spare axillary lymph node dissection for the patient in the operating room.

Predicting the Status of the Nonsentinel Lymph Nodes in early Breast Cancer Patients with Positive Sentinel Lymph Nodes / 한국유방암학회지

PURPOSE: The need for completion axillary lymph node dissection, even in early breast cancer patients with a positive sentinel lymph node, has been questioned. The purpose of this study was to determine the factors that predict the presence of metastasis in non-sentinel lymph nodes (NSLNs) when the sentinel lymph node (SLN) was positive. METHODS: Between December 1998 and June 2004, the records of 104 early breast cancer patients with a positive SLN and who underwent completion axillary lymph node dissection were reviewed. The clinicopathological features in SLN-positive patients were evaluated as possible predictors of metastatic NSLN. RESULTS: Forty four (42.3%) of the 104 patients with positive SLN had metastatic NSLNs. In a univariate analysis, unicentric multifocality (P=0.016), lymphovascular invasion (P=0.006) and SLN metastasis larger than 2 mm (P= 0.003) were associated with positive NSLN findings. The number of SLNs removed was significantly associated as a negative predictor (P=0.043). A multivariate analysis revealed that SLN metastasis >2 mm (P=0.021) and lymphovascular invasion (P=0.040) were independent predictors of metastatic NSLN. CONCLUSION: The likelihood of metastatic NSLNs correlates with the size of the largest SLN metastasis and the presence of lymphovascular invasion of the primary tumor. Even though in early breast cancer with positive SLNs, incorporating these factors may help determining which patients would benefit from additional axillary lymph node dissection.

Clinical Experience for Sentinel Lymphadenectomy Alone in Early Breast Cancer / 한국유방암학회지

PURPOSE: This study was performed to evaluate the efficacy and safety of sentinel lymph node biopsy alone (SLNB) without axillary lymph node dissection. METHODS: Between Jun. 1999 and Dec. 2002, we carried out SLNB in 85 consecutive patients with T1 breast cancer whose sentinel lymph node(s) were tumor-free on intra- operative frozen section. Even when sentinel lymph node(s) turned out to be positive by permanent pathology, additional axillary lymph node dissection was not performed. Patients underwent total mastectomy or partial mastectomy and received an appropriate adjuvant therapy according to the characteristics of the primary tumor. All patients who had breast conserving surgery received postoperative radiotherapy to the remaining breast, but not to the axilla. RESULTS: SLNB only took 14 minutes and yielded no postoperative complications. Among the 85 patients whose sentinel lymph nodes were tumor-free on frozen section, 11 patients were diagnosed as metastatic in the permanent pathology. One of them had a macro-metastasized (2.5 cm) sentinel lymph node, and the rest had micro-metastasized sentinel lymph nodes. For 23.1 months of mean follow-up period, all the patients including a patient who died of liver metastases at 17 months showed no evidence of axillary recurrence. CONCLUSION: No axillary recurrence following SLNB suggests that SLNB may be a good alternative to routine axillary lymph node dissection while providing less surgical morbidity in women with a small breast cancer. However, more patients accumulation and follow-up period will be needed for the final conclusion.

The Necessity of Learning Period for Sentinel Lymphadenectomy in Breast Carcinoma / 한국유방암학회지

PURPOSE: Although an axillary lymph node dissection (ALND) has been considered as an ultimate procedure for axilla in the breast carcinoma, complications after ALND and conceptual revolution for the role of axillary nodes have made it necessary to look for an alternative. Recent studies of sentinel lymphadenectomy (SLND) have shown that SLND accurately predict axillary nodal status. However, for a satisfying outcome in SLND, a learning period would be required, as other surgical procedures do. In this study, the necessity of the learning period for SLND were examined. METHODS: From Nov. 1998 to Dec. 2001, 178 patients with invasive breast carcinoma were treated with SLND simultaneously followed by ALND. The period for the first 54 patients, Nov. 1998 to May. 1999, was set as a 'learning period'. Differences of the detection rate and the false negative rate for the 'learning period' and 'after the learning period' were compared. Also changes in the detection rate and the false negative rate with the accumulated experiences for SLND were evaluated. RESULTS: The sentinel lymph nodes were not identified in 5 patients through the whole period. Three of them occurred in the 'learning period' (5.6%, 3/54) and the rest occurred 'after the learning period' (1.6%, 2/124)(P=0.04). The false negative rate was 16.7% (4/24) in the 'learning period' and 0.0% in 'after the learning period' (P=0.00). The detection rate and the false negative rate improved with the accumulation of experiences for SLND. CONCLUSION: It is certain that 'learning period' for SLND is crucial. During this period, the improvement and stabilization of this skill is achieved.

Experiences with Sentinel Lymphadenectomy in 157 Cases of Breast Carcinoma / 한국유방암학회지

PURPOSE: The status of the axillary nodes in breast carcinoma patients is one of the most significant prognostic factors. Although many trials in order to gain information concerning axillary status, to date there is no satisfactory procedures other than axillary lymph node dissection (ALND) which has many complications. Recently sentinel lymphadenectomy has become accepted as an alternative to ALND. METHODS: Sentinel lymphadenectomy followed by ALND was performed in 157 patients with invasive breast carcinoma between Nov. 1998 and Dec. 2000. The period from Nov. 1998 to May. 1999, which included the first 54 patients was set as a learning period. 99m Tc antimony trisulfide colloid was used in the detection of the sentinel node, with a gamma camera preoperatively and then a gamma probe in the operating room. The dissected sentinel nodes were examined by both frozen section and serial section (SS) with immunohistochemical stains (IHC). RESULTS: The detection rate was 96.8%. The false negative rate was 22.6% for the frozen sections and 4.8% for the permanent sections. The detection rate and the false negative rate had improved after the learning period with statistic significance (P-value<0.05). The false negative rate for tumors less than 2 cm was significantly lower than that for tumors greater than 2 cm (P-value=0.000). CONCLUSION: Sentinel lymphadenectomy using 99m Tc anti-nomy trisulfide colloid showed high detection rate and low false negative rate. The learning period is thought to be necessary for its clinical application. In the future, sentinel lymphadenectomy may replace axillary lymph node dissection in certain subgroups of breast carcinoma patients.

Experiences with Sentinel Lymphadenectomy in 157 Cases of Breast Carcinoma

PURPOSE: The status of the axillary nodes in breast carcinoma patients is one of the most significant prognostic factors. Although many trials in order to gain information concerning axillary status, to date there is no satisfactory procedures other than axillary lymph node dissection (ALND) which has many complications. Recently sentinel lymphadenectomy has become accepted as an alternative to ALND. METHODS: Sentinel lymphadenectomy followed by ALND was performed in 157 patients with invasive breast carcinoma between Nov. 1998 and Dec. 2000. The period from Nov. 1998 to May. 1999, which included the first 54 patients was set as a learning period. 99m Tc antimony trisulfide colloid was used in the detection of the sentinel node, with a gamma camera preoperatively and then a gamma probe in the operating room. The dissected sentinel nodes were examined by both frozen section and serial section (SS) with immunohistochemical stains (IHC). RESULTS: The detection rate was 96.8%. The false negative rate was 22.6% for the frozen sections and 4.8% for the permanent sections. The detection rate and the false negative rate had improved after the learning period with statistic significance (P-value<.05). The false negative rate for tumors less than 2 cm was significantly lower than that for tumors greater than 2 cm (P-value=0.000). CONCLUSION: Sentinel lymphadenectomy using 99m Tc antinomy trisulfide colloid showed high detection rate and low false negative rate. The learning period is thought to be necessary for its clinical application. In the future, sentinel lymphadenectomy may replace axillary lymph node dissection in certain subgroups of breast carcinoma patients.

Sentinel Lymph Node Biopsy Using Radioactive Material in Breast Cancer Patients

PURPOSE: In breast cancer, the single most important prognostic indicator is the axillary nodal status of the metastasis. This study was designed to discern the accuracy of sentinel node biopsy using radioactive material for the prediction of axillary nodal status. METHODS: The study group was selected from the Department of Surgery at Yonsei University Yongdong Severance hospital, and comprised 54 patients diagnosed with breast cancer with clinical stages I or II from Nov. 1998 to May. 1999. We performed lymphoscintigraphy and gamma-probe with 99mTc-antimony trisulfide colloid for the detection of sentinel nodes. The radiotracer was injected peritumorily 2 hours prior to the operation. During, surgery and after the sentinel node biopsy was performed, level I and II axillary lymph node dissection was completed. The sentinel node was divided into 3 pieces evenly. One piece was tested for frozen section diagnoses and the others were fixed for H&E staining and immunohistochemistry (IHC) using cytokeratin antibody to detect metastasis. RESULTS: The mean number of the sentinel nodes was 1.59. The detection rate of the sentinel node was 94.4% (51/54). Among 35 patients with negative sentinel nodes on frozen section, 4 patients (11.4%) demonstrated tumor metastasis on both permanent serial H&E sections and IHC using cytokeratin. Of the 31 patients with negative sentinel node by permanent serial section, 4 patients displayed metastasis in nonsentinel axillary nodes. These results revealed that thesensitivity of the sentinel node biopsy was 83.3% and the negative predictive value was 81.1%. The false negative rate of the sentinel node biopsy was 16.7% with the permanent H&E and IHC section and 33.3% with the frozen section. In particular, in cases with tumors less than 2 cm, the sensitivity was 100% in frozen section as well as permanent section. CONCLUSION: A combination of preoperative lymphoscintigraphy and gamma probe-guided methods achieves high success rates in the identification of sentinel nodes. Multiple serial sections and IHC of the nodes are required to detect the micrometastasis of the sentinel nodes. The sentinel node biopsy is significantly accurate and may be considered as an alternative to replace the routine axillary node dissection, particularly in T1 breast cancer patients.

Sentinel lymph node biopsy using radioactive material in breast cancer patients / 한국유방암학회지

BACKGROUND: In breast cancer, the single most important prognostic indicator is the axillary nodal status of the metastasis. The sentinel lymph nodes are the lymph nodes of the primary tumor that is drained first, which can predict the presence of the tumor metastasis with high accuracy. This study was designed to find out how accurately the sentinel node biopsy using radioactive material can predict axillary nodal status. METHODS: The patients were selected from the department of surgery at Yonsei University Yongdong Severance hospital, consisting of 54 patients with breast cancer diagnosed as clinical stages I or II from Nov. 1998 to May, 1999. We performed lymphoscintigraphy and gamma-probe method with 99mTc-antimony trisulfide colloid for detection of sentinel nodes. The radiotracer was injected around the tumor 2 hours prior to the operation. In the operating room, after the sentinel node biopsy had been performed, levels II and II axillary lymph node dissection were completed. The sentinel node was divided into 3 pieces evenly. One piece was tested for frozen section diagnoses and the others were fixed in 10% formalin for H&E staining and immunohistochemistry(IHC) using cytokeratin antibody to detect metastatic tumor cells. RESULTS: The mean size of the primary tumor was 2.51cm in diameter and the mean number of the sentinel nodes was 1.59. The detection rate of the sentinel node was 94.4%(51/54). Among 35 patients with negative sentinel nodes on frozen section, 4 patients(11.4%) had tumor metastasis on both permanent serial H&E sections and IHC using cytokeratin. Out of the 31 patients with negative sentinel node by permanent serial section, 4 patients had tumor metastasis in nonsentinel axillary nodes. These results revealed that the sensitivity of the sentinel node biopsy was 83.3% and the negative predictive value to predict the axillary node status was 81.1%. The false negative rate of the sentinel node was 16.7% by the permanent H&E and IHC section and 33.3% by the frozen section. Especially, in casese with tumors loss than 2cm, the sunsitivity was 100% in frozen section as well as permanent section. CONCLUSION: A combination of preoperative lymphoscintigraphy and gamma probe-guided methods achieve high success rates in the identification of sentinel nodes. Multiple serial sections and IHC of the nodes are required to detect the micrometastasis of the sentinel nodes. The sentinel node biopsy is significantly accurate and may be considered as an alternative to replace the routine axillary node dissection, especially in T1 breast cancer patients.