Immunohistochemical stains fail to increase the detection rate of micrometastatic melanoma in completion regional lymph node dissection specimens

In melanoma patients, examination of tissue sections stained for immunohistochemical markers as an adjunct to examination of haematoxylin and eosin (H&E)-stained sections has been shown to increase the detection rate of micrometastatic disease in sentinel lymph nodes (SLNs). However, immunohistochemical stains are not routinely performed on completion regional lymph node dissection (CLND) specimens in most centres and it is not known whether their use would increase the detection of micrometastatic disease in these specimens. This study was performed to determine whether the application of immunohistochemical stains for S100 and HMB45 (in addition to H&E stains) increases the detection of micrometastatic disease in CLND specimens of melanoma patients and whether their use would be cost-effective in routine pathological practice. Forty-nine CLND specimens from patients with a prior positive SLN biopsy were examined by performing H&E stains and immunohistochemical stains for S100 protein and HMB-45 on each node, and the detection rate of melanoma metastases using H&E-stained sections was compared with that using immunohistochemically stained sections. The number of nodes in the CLND specimens ranged from 4 to 37 (median 14, mean 14.7). Nodal deposits of melanoma cells were detected in 12 of 49 cases (24%). Among these 12 positive cases, the mean number of positive nodes per CLND specimen was 2.2 (range, 1-14). The total number of positive nodes in the 12 CLND specimens was 27, accounting for 3.8% of the 720 nodes removed in the study group of 49 cases. Melanoma cells in all 27 positive nodes were identified both on the H&E-stained slides and the slides stained immunohistochemically for S100. The melanoma cells were positive for HMB45 in 24 of the 27 lymph nodes that contained metastatic melanoma (the metastatic melanoma cells were negative for HMB45 in three positive nodes from one specimen). No further positive lymph nodes were detected with the immunostains that had not been identified on the H&E-stained sections. This study suggests that the use of immunostains does not increase the detection rate of metastatic melanoma in CLND specimens, and that their routine use would not be cost-effective. We therefore recommend only H&E staining on sections of all lymph nodes in CLND specimens from melanoma patients.     

The sentinel node in colorectal carcinoma. Mapping technique,pathologic assessment,and clinical relevance

One of the most important prognostic factors in colorectal cancer is the presence or absence of regional lymph node metastases. In many instances, micrometastatic disease may not be found on routine pathologic analysis using hematoxylin and eosin staining, but may be discovered only with immunohistochemical methods or polymerase chain reaction assay. Lymphoscintigraphy with biopsy of the sentinel nodes, defined as the first nodal basin in the drainage pathway of a tumor, was developed to provide accurate staging without the morbidity associated with the classic lymph node dissections performed for melanoma or breast cancer. This concept has recently been applied to colorectal cancers, but the method used is unique because oncologic principles of resection are still adhered to for the primary tumor along with en bloc resection of the locoregional mesenteric nodes, some of which are sentinel nodes. Sentinel nodes are ideal for sensitive pathologic techniques of detecting micrometastatic disease, as they often reflect the status of the entire locoregional nodal basin. Gross metastatic nodes reveal significant prognostic information and guide the use of adjuvant therapy in affected patients. However, the detection of micrometastatic disease in sentinel nodes by sensitive pathologic methods has not been proven to result in poor prognosis or benefit from adjuvant therapy for colorectal cancer.     

The role of preoperative ultrasonography in reducing the number of sentinel lymph node procedures in melanoma

Sentinel lymph node (SLN) biopsy is the most effective method to nodally stage patients with melanoma. However, SLN metastases are an indication for a complete regional lymphadenectomy. The aim of this study was to evaluate the ability of ultrasound (US) and US-guided fine needle aspiration biopsy (US-FNAB) to reduce the number of patients requiring a second surgical procedure. Fifty-seven patients with melanoma underwent preoperative US of the regional lymph nodes before SLN biopsy. In patients with US malignant lymph nodes, US-FNAB was performed. Only patients with cytologically proven lymph node metastases proceeded directly to a complete regional lymphadenectomy, whereas, in all others, SLN biopsy was performed. Fourteen patients (25%) had metastases in the regional lymph nodes. There were 40 benign and 17 malignant US results. US-FNAB was performed in 14 patients. It was positive in three, negative in nine and inadequate sampling was obtained in two. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of US were 71%, 84%, 59% and 90%, respectively. US of the regional lymph nodes with US-FNAB enables the safe selection of patients who should proceed directly to a complete regional lymphadenectomy. However, the sensitivity and PPV of the method are low.     

Sentinel node localisation: A new prospective in the treatment of nodal melanoma metastases.

Sentinel node (SN) mapping and biopsy is a procedure that accurately stages the regional lymph node (LN) basin. Defined patterns of lymphatic drainage allow intraoperative determination of the first (sentinel) lymph node in the regional basin, and the absence of metastatic disease in the SN accurately reflects the absence of melanoma in the remaining regional nodes. The use of radiocolloid and a hand-held gamma detecting probe (GDP) together with a vital blue dye provides optimal results, and allows for the successful identification of the SN in over 99% of the procedures. Close collaboration between surgeons, nuclear radiologists and pathologists is required to ensure optimal results. Examination of serially sectioned SNs by hematoxylin-eosin staining (H&E), immunohistochemical staining and perhaps in the near future RT-PCR should reduce the number of patients with missed microscopic melanoma in the regional lymph nodes. Furthermore, the survival benefit recently reported in patients with melanoma metastatic to regional nodes using high dose of interferon alpha-2b signals that the surgeons should aggressively examine patients for the presence of occult regional melanoma metastases. Intraoperative SN mapping and SN biopsy are cost-effective procedures that allows accurate identification of regional lymph nodes that contain metastatic melanoma.     

The role of sentinel lymph node biopsy for melanoma

Regional lymph nodes are a common site of melanoma metastases, and the presence or absence of melanoma in regional lymph nodes is the single most important prognostic factor for predicting survival. Furthermore, identification of metastatic melanoma in lymph nodes and excision of these nodes may enhance survival in a subgroup of patients whose melanoma has metastasized only to their regional lymph nodes and not to distant sites. Sentinel lymph node (SLN) biopsy was developed as a low morbidity technique to stage the lymphatic basin without the potential morbidity of lymphedema and nerve injury. The presence or absence of metastatic melanoma in the SLN accurately predicts the presence or absence of metastatic melanoma in that lymph node basin. When performed by experienced centers, the false-negative rate of SLN biopsy is very low. As such, the nodal basin that contains a negative SLN will usually be free of microscopic disease. Since occult micrometastatic disease affects only 12% to 15% of patients with melanoma, selective SLN dissection allows up to 85% of patients with melanoma to be spared a formal lymph node dissection, thus avoiding the complications usually associated with that procedure. While standard pathologic evaluation of lymph nodes may miss metastatic melanoma cells, more sensitive techniques are developing which may identify micrometastases more accurately. The clinical significance of these micrometastases remains unknown and is the subject of active investigations.     

A case of malignant melanoma, axillary lymph node metastasis proved by fine needle aspiration biopsy cytology

We report the case of a 57-year-old man with a malignant melanoma in the subunguinal region of his right thumb associated with the palpable axillary lymph nodes. Its metastasis was confirmed by fine needle aspiration biopsy cytology. Prognosis if a malignant melanoma is protean by its stage. Thus information regarding the axillary metastasis is mandatory for deciding the therapeutic plan. Fine needle aspiration biopsy cytology is a useful technique for diagnosing lymph node metastasis.     

Aspiration biopsy and the clinical management of patients with malignant melanoma and palpable regional lymph nodes

BACKGROUND: The presence of lymph node metastases in patients with malignant melanoma implies a significant decrease in survival. The authors investigated the efficacy of fine-needle aspiration biopsy (FNAB) in the diagnosis of metastatic malignant melanoma in 115 patients with melanoma and clinically suspicious regional lymph nodes. METHODS: One hundred thirty-three FNABs were performed by cytopathologists after referral from surgeons or oncologists using a 25-gauge or 27-gauge needle. RESULTS: The cytologic diagnosis was negative in 35, atypical in 1, suspicious in 2, and positive for malignant melanoma in 95. Regional lymph node dissections were performed in 78 patients. Of these, 70 positive FNABs were confirmed with no false-positive results. The atypical FNAB was proven positive for malignant melanoma at surgery. Of the two suspicious FNABs, one was confirmed as positive and one showed dermatopathic lymphadenopathy. Of the 35 negative FNAB specimens, 5 patients underwent surgery; 3 FNABs were found to be negative and 2 FNABS were falsely negative. Twenty patients with negative aspirates were followed clinically for 22-45 months (mean, 32 months); 19 patients had no evidence of disease and 1 patient died of disseminated melanoma. CONCLUSIONS: FNAB of palpable lymphadenopathy in patients with malignant melanoma can provide a rapid and accurate assessment of lymph node status and expedite the therapeutic management of these patients.     

Melanoma trials that defined surgical management

Treatment of regional lymph nodes in melanoma has been controversial for more than a century. A series of clinical trials evaluating elective lymph node dissection and then sentinel lymph node biopsy have helped define the current standard of care. These trials resulted in increasingly selective application of surgical intervention for regional lymph nodes in melanoma. First by focusing on optimal candidates for elective lymph node dissection and then by identifying patients through sentinel lymph node biopsy. The current standard of sentinel lymph node biopsy for appropriately selected patients and nodal observation for many patients, even with involved sentinel nodes is both more accurate in staging and much less morbid than what came before.     

Lower tumour burden and better overall survival in melanoma patients with regional lymph node metastases and negative preoperative ultrasound

Background

The purpose of the study was to evaluate the ability of ultrasound (US) and fine needle aspiration biopsy (FNAB) in reducing the number of melanoma patients requiring a sentinel node biopsy (SNB); to compare the amount of metastatic disease in regional lymph nodes in SNB candidates with clinically uninvolved lymph nodes and of those with US uninvolved lymph nodes; and to compare the overall survival (OS) of both groups.

Methods

Between 2000 and 2007, a SNB was successfully performed in 707 patients with melanoma. The preoperative US of the regional lymph node basins was performed in 405 SNB candidates. In 14 of these patients, the US-guided FNAB was positive and they proceeded directly to lymph node dissection. In 391 patients, the preoperative US was either negative (343 patients) or suspicious (48 patients) (US group). In the remaining 316 patients the preoperative US was not performed (non-US group).

Results

The proportion of macrometastatic sentinel lymph nodes (SN), number of metastatic lymph nodes per patient and proportion of nonsentinel lymph node metastases were found to be lower in the US group compared to the non-US group. The smaller tumour burden of the US group was reflected in a significantly better OS of patients with SN metastases.

Conclusions

The preoperative US of regional lymph nodes spares some patients with melanoma from undergoing a SNB. Patients with regional metastases and a negative preoperative US have a significantly lower tumour burden in comparison to those with clinically negative lymph nodes, which is also reflected in a better OS.     

Ultrasound assessment of lymph nodes in the management of early-stage melanoma

It has been suggested that ultrasound examination of regional lymph nodes provides staging and prognostic information that is as accurate as sentinel node biopsy in patients who present with cutaneous melanoma. However, in most studies the sensitivity of ultrasound in detecting nodal metastasis has been low (23% in a recent large series). For the present, sentinel node biopsy remains the most accurate method of regional node staging in patients with newly diagnosed melanoma.     

Surgical management of primary cutaneous melanoma: excision margins and the role of sentinel lymph node examination

Surgical strategies for managing patients who have primary cutaneous melanoma have changed dramatically over the past 30 years. More conservative excision margins have been shown to be adequate, and routine complete lymph node dissection (CLND)has been abandoned since the sentinel node (SN) biopsy technique was introduced. Knowledge of a patient's SN status not only provides a reliable guide to prognosis, but also allows CLND to be avoided in 80% to 85% of patients. Recent clinical trial results suggest that SN biopsy, with immediate CLND if an SN is positive,confers a survival advantage in those who have metastatic disease in regional nodes. Minimally invasive and noninvasive methods of SN assessment, such as magnetic resonance spectroscopy, are being evaluated.     

早期乳腺癌前哨淋巴结活检的前沿进展

医学影像技术的飞速发展提高了早期乳腺癌腋窝淋巴结影像学诊断的能力,能够检测到腋窝淋巴结更早、更小的转移灶,通过影像学引导下穿刺活检明确淋巴结的转移状况,从而为患者提供更完善的术前腋窝分期。对于术前超声影像学检查为腋窝淋巴结阴性的患者是否可以避免术中腋窝前哨淋巴结活检,目前有两项大型临床试验正在进行中。山东大学附属山东省肿瘤医院乳腺病中心通过微泡超声造影显像技术引导腋窝前哨淋巴结术前穿刺活检,正在探索一种高效、准确和超微创的腋窝前哨淋巴结活检技术。术前影像学检查大大提高了临床腋窝淋巴结阳性患者的检出率,但也让一部分患者失去了通过腋窝前哨淋巴结活检替代腋窝淋巴结清扫的机会,对临床查体淋巴结阴性但超声发现可疑淋巴结并通过穿刺活检确诊转移的患者是否适合前哨淋巴结活检,本文也进行了相关的探讨和研究。另外,对内乳前哨淋巴结活检的相关研究结果进行了汇总。     

Comparison of positron emission tomography scanning and sentinel node biopsy in the detection of micrometastases of primary cutaneous malignant melanoma.

PURPOSE: Sentinel node biopsy (SNB) is a surgical technique for detecting micrometastatic disease in the regional draining nodes. 2-fluorine-18-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) scanning is an imaging technique that can detect clinically undetectable metastases. This prospective study was undertaken to compare the sensitivity of FDG-PET scanning with SNB in the detection of micromatastatic malignant melanoma. PATIENTS AND METHODS: Fifty consecutive patients (23 women, 27 men; mean age, 53 years) with primary melanoma >1 mm thick or lymphatic invasion were recruited (mean, 2.41 mm). Primary lesions had been narrowly excised (<1 cm). Patients underwent PET scanning followed by SNB, using a dual technique. Preoperative lymphoscintigraphy was used to identify the draining basin. Lymph nodes were examined histologically and immunostained for S100 and HMB 45. RESULTS: The sentinel node (SN) was identified in all patients. Fourteen patients (28%) had positive SNBs, including eight patients with melanoma <1.5 mm thick. In none of these 14 patients did PET scans identify metastatic disease in the SN or draining basin. In seven patients, the PET scans were positive in other locations, and in four cases, this was suspicious of metastatic disease. However, no patient has developed recurrent melanoma (mean follow-up, 15 months). All patients with positive SNBs underwent therapeutic lymph node dissection. Further lymph node involvement was found in two patients (primary lesions, 1.3 mm and 3.5 mm thick). CONCLUSION: This study demonstrates the limitations of FDG-PET scanning in staging patients with primary melanoma. SNB is the only reliable method for identifying micrometastatic disease in the regional draining node.     

Ki-67 expression in melanoma. A potential method of risk assessment for the patient with a positive sentinel node

Melanoma patients with a positive sentinel node biopsy generally proceed to regional lymph node dissection, though ultimately only around 20% have evidence of tumour in their "non-sentinel" nodes. A means to identify patients at high risk of non-sentinel node involvement could potentially spare a large number of patients a procedure with significant morbidity. The proliferation marker Ki-67 has been associated with tumour progression in primary melanoma but has not been extensively studied in metastases. The study aims to investigate Ki-67 in primary melanoma and lymph node metastases and investigate any relationship with disease progression. Tissue Arrays of primary melanoma (n=79) and lymph node metastases (n=92) were constructed from paraffin embedded tissue and Ki-67 expression examined by immunohistochemistry. Staining positivity and intensity were assessed and correlated with standard staging criteria and clinical outcome. High Ki-67 expression was associated with both Breslow thickness (chi(2)=8.54, p=0.035) and presence of ulceration (Fisher's Exact test p=0.003) in primary melanoma. In lymph node metastases high Ki-67 expression correlated with Nodal (N) Stage (chi(2)=8.193, p=0.0 17). High Ki-67 expression is associated with melanoma progression and multiple lymph node involvement. This might potentially form the basis of a risk analysis for patients with positive sentinel nodes.     

Occult axillary lymph node metastases are of no prognostic significance in breast cancer

The significance of occult metastases in axillary lymph nodes in patients with carcinoma of the breast is controversial. Additional sections were cut from the axillary lymph nodes of 477 women with invasive carcinoma of the breast, in whom no metastases were seen on initial assessment of haematoxylin and eosin stained sections of the nodes. One section was stained with haematoxylin and eosin, and one using immunohistochemistry with two anti-epithelial antibodies (CAM5.2 and HMFG2). Occult metastases were found in 60 patients (13%). The median follow-up was 18.9 years with 153 breast cancer related deaths. There was no difference in survival between those with and those without occult metastases. Multivariate analysis, however, showed that survival was related to tumour size and histological grade. This node-negative group was compared with a second group of 202 patients who had one involved axillary node found on initial assessment of the haematoxylin and eosin sections; survival was worse in the patients in whom a nodal metastasis was found at the time of surgery. Survival was not related to the size of nodal metastases in the occult metastases and single node positive groups. Some previous studies have found a worse prognosis associated with occult metastases on univariate analysis, but the evidence that it is an independent prognostic factor on multivariate analysis is weak. We believe that the current evidence does not support the routine use of serial sections or immunohistochemistry for the detection of occult metastases in the management of lymph node negative patients, but that the traditional factors of histological grade and tumour size are useful.     

Immunohistochemistry with pancytokeratins improves the sensitivity of sentinel lymph node biopsy in patients with breast carcinoma

BACKGROUND: Sentinel lymph node (SLN) biopsy is being investigated as a staging procedure for breast carcinoma. The authors evaluated whether immunohistochemical (IHC) analysis improves the sensitivity of this procedure. METHODS: Forty-four women with breast carcinoma were recruited for SLN biopsy. Preoperative lymphoscintigraphy was followed by intraoperative localization using a handheld gamma probe and blue dye. After SLN identification, an immediate complete axillary lymph node dissection was performed in all patients. All lymph nodes were subjected to routine histology (hematoxylin and eosin [H&E]) and IHC using antibody to cytokeratins. RESULTS: The SLN was identified in 41 of 43 patients (95%). Successful SLN identification was independent of biopsy technique (open surgical [95%] vs. fine-needle aspiration/core needle biopsy [96%]). Twelve of 41 patients (29%) had evidence of lymph node metastasis in the SLN by routine histology. Of the twenty-nine patients with H&E negative SLN, 3 were found to have metastasis by IHC for a conversion rate of 10%. Fifteen of 41 patients (37%) had evidence of metastasis in SLN. All 26 patients with H&E and IHC negative SLN had negative nonsentinel lymph nodes by routine histology and IHC (100% negative predictive value). All patients with tumors < 2 cm and micrometastasis to the SLN had no additional lymph node disease, in contrast to patients with lesions > 2 cm or patients with macrometastasis to the SLN (P = 0.007). CONCLUSIONS: These results confirm that SLN biopsy is extremely accurate for patients with breast carcinoma, even after open surgical biopsy. IHC analysis or serial sectioning of SLN improves the sensitivity of this staging technique.     

Update on the use of sentinel node biopsy in patients with melanoma: who and how

This review highlights the utility of sentinel lymph node biopsy for melanoma. The histopathologic status of the sentinel lymph node is acknowledged as the most powerful indicator of prognosis. Changes by the American Joint Committee for Cancer have not only incorporated the importance of ulceration in cutaneous melanoma, but have focused on the number of metastatic lymph nodes as well as whether the disease is macro- or micrometastatic. Specific technical aspects of the sentinel lymph node procedure are discussed, with emphasis on practical guidelines to minimize incorrect nodal staging.     

Minimal risk of macrometastases in the non-sentinel axillary lymph nodes in breast cancer patients with micrometastatic sentinel lymph nodes and preoperatively ultrasonically uninvolved axillary lymph nodes

Micrometastases in the sentinel lymph node (SLN) carry a considerable risk of macrometastases in the non-sentinel lymph nodes (NSLN), resulting in axillary lymph node dissection (ALND). Preoperative ultrasound (US) examination of the axillary lymph nodes combined with a fine-needle aspiration biopsy (FNAB) has been proved to discover metastases in the axillary lymph nodes. The aim of our study was to assess the risk of macrometastases in NSLN in patients with micrometastatic SLN after a preoperative US examination of the axillary lymph nodes. The study included 36 patients in whom, after preoperative axillary US, micrometastases in the SLN were revealed and ALND was subsequently performed. At final histopathology, no macrometastases were discovered in the NSLN. In four patients, additional micrometastases were discovered in the NSLN. In conclusion, the risk of macrometastases in the NSLN in patients with preoperatively ultrasonically uninvolved axillary lymph nodes is minimal.     

早期乳腺癌前哨淋巴结示踪和定位的临床研究

背景与目的:目前乳腺癌手术都常规行腋窝淋巴结清扫(axillarylymphnodedissection,ALND)。对早期乳腺癌行ALND能否提高患者的生存率尚有争论,但其带来的并发症却明确。对早期乳腺癌能否用前哨淋巴结活检(sentinellymphnodebiopsy,SLNB)来代替ALND是目前国际研究的热点。本研究旨在探讨用不同的方法示踪和定位行乳腺癌SLNB的可行性及其临床意义,并探索多层切片加免疫组化技术对判断前哨淋巴结(sentinellymphnode,SLN)微转移灶的意义。方法:对121例早期乳腺癌患者行SLN示踪活检,其中美蓝法38例,单纯用美蓝标记;联合法83例,用美蓝和99mTc-SC同时标记。术后将SLN和所有常规ALND的淋巴结作常规病理检查,对阴性的SLN作3层切片加免疫组化检查,以检测淋巴结的微转移灶。结果:美蓝组和联合组活检的成功率分别为81.6%和95.2%;对腋窝淋巴结状况预测的准确率分别为93.5%和97.5%。淋巴结闪烁成像(lymphoscintigraphy,LS)和手持式γ-探头术前定位SLN的成功率分别为23%和92%(P<0.05)。本组194枚常规病理检查阴性的SLN,后经3层切片加免疫组化检查,发现13例21枚(11%)SLN含微转移灶。再用联合法检查对腋窝淋巴结预测的准确率上升为98.7%;假阴性率下降为3.2%。结论:美蓝法和联合法行SLNB均能准确地反映腋窝淋巴结状况,但联合法相对较好。LS对SLNB的意义尚待进一步探讨。多层切片加免疫组化检查能提高SLN微转移灶的发现率。