Prognostic value of RT-PCR tyrosinase detection in peripheral blood of melanoma patients

Malignant melanoma (MM) prognosis has been related to tumour thickness and clinical stage and metastasis risk has been associated with presence of tumour cells in peripheral blood. The aim of this study was to determine the relationship between presence of tyrosinase in peripheral blood of MM patients and their clinical prognosis. Blood samples from 58 MM patients (stage I-IV) were analysed, using RT-PCR assay to detect tyrosinase mRNA. The results showed that positive RT-PCR assay for tyrosinase were significantly associated with clinical status and tumour thickness. After a median follow-up of 24 months, RT-PCR results were found to be significant correlated with recurrence (p<0.05) and clinical stage III (p<0.05). Separate analysis of stage III tumours to determine the prognostic value of tyrosinase presence in peripheral blood showed an overall 24-month survival rate of 70% in the RT-PCR negative group versus 10% in the positive group (p<0.02). These results suggest that detection of circulating melanoma cells may be especially relevant in stage III patients, in whom RT-PCR positivity defines a subpopulation at high risk of recurrence.     

Immunoreactivity with the anti-MAGE antibody 57B in malignant melanoma: frequency of expression and correlation with prognostic parameters.

The melanoma-associated antigen (MAGE) family consists of a number of antigens initially recognized by cytotoxic T lymphocytes, which are currently being investigated for immunotherapy of patients with metastatic melanoma and other tumor types. Expression of MAGE mRNA in melanocytic tumors is said to be restricted to invasive malignant tumors and absent in nevi. Recently, a monoclonal antibody (57B) has become available to examine MAGE protein expression in archival material. In this study, we performed immunohistochemical analysis on 132 melanocytic nevi and 205 melanomas (85 primary cutaneous melanomas and 120 metastatic tumors) to determine the frequency of MAGE expression and to explore a potential correlation with various prognostic parameters. None of the melanocytic nevi and none of the 20 in situ melanomas was immunopositive with the antibody 57B. Immunoreactivity was present in 17 of 65 (26%) primary invasive melanomas of the skin and in 30 of 120 (25%) metastatic tumors. Positive immunostaining did not correlate with tumor stage (P = .66), Breslow thickness (P = .39), Clark level (P = .5), or the histologic type of melanoma (P = .23) but was associated with a brisk infiltrate of lymphocytes involving the vertical growth phase of melanomas (P = .01). Because tumor-infiltrating lymphocytes in melanoma are associated with longer survival, our findings suggest a potential prognostic role for MAGE. Furthermore, the seeming restriction of immunopositivity to invasive malignant tumors suggests a potential diagnostic role for the antibody 57B in confirming the malignant potential of a melanocytic tumor.     

Tumor lymphangiogenesis predicts melanoma metastasis to sentinel lymph nodes.

Cutaneous melanoma is a common melanocytic neoplasm that can quickly metastasize to regional lymph nodes. Currently, prognosis is determined by measuring tumor thickness but more reliable markers for metastatic spread are urgently needed. We investigated whether the extent of tumor lymphangiogenesis can predict melanoma metastasis to sentinel lymph nodes. We quantified the extent of tumor lymphangiogenesis, as well as other factors, in excised primary tumors and in sentinel lymph node biopsy samples from 45 patients with primary cutaneous melanoma. The results were correlated with histological and clinical outcome. Primary melanomas from patients whose tumors had metastasized to the sentinel lymph nodes contained prominent 'hot spots' of increased lymphatic vessel density, compared to nonmetastatic tumors. Multivariate risk analysis revealed that the lymphatic vascular area of primary melanomas, an index of tumor lymphangiogenesis, was the most sensitive prognostic marker for sentinel lymph node metastasis, and was even able to more accurately predict which tumors were metastatic to sentinel lymph nodes than the currently used method of measuring tumor thickness. Highly lymphangiogenic melanomas maintained their lymphangiogenic activity after metastasis to the sentinel lymph node. The extent of tumor lymphangiogenesis is a highly sensitive (83%) and specific (89%) prognostic marker of lymph node metastasis. Assessment of lymphangiogenesis in primary melanomas may be a more effective approach than the currently used technique of measuring tumor thickness in selecting patients with early metastatic disease for aggressive therapy.     

Prognostic significance of the histological features of malignant melanoma

A review of 694 patients with localized cutaneous malignant melanoma (clinical stage I) revealed that three histological features of the primary lesion had no effect of their own on survival rate but derived their prognostic significance only because of their close correlation with tumour thickness. Primary lesions of superficial spreading histogenetic type, or of low mitotic activity or showing evidence of partial regression appeared to have a more favourable prognosis than lesions of nodular histogenetic type or of high mitotic activity or showing no regression. However, the former three histological features were predominant in thin lesions which had a better prognosis than thicker lesions. It was concluded that these features exerted only an indirect effect upon survival, tumour thickness being the most important prognostic determinant.     

Patterns of melastatin mRNA expression in melanocytic tumors

The melanocyte-specific gene Melastatin (MLSN1) shows an inverse correlation of mRNA expression with metastatic potential in human and murine cell lines in vitro. Melastatin mRNA expression in primary cutaneous melanoma also has been found to correlate with disease-free survival. The histologic patterns of Melastatin mRNA expression in nevi, primary melanoma, and melanoma metastases have not been described previously. Using in situ hybridization with (35)S-labeled probes, we examined Melastatin mRNA expression in 64 cases of normal skin, benign melanocytic nevi, primary cutaneous melanomas, and melanoma metastases. Ubiquitous melanocytic expression of Melastatin mRNA was observed in all benign melanocytic proliferations (14 of 14), although some nevi showed a gradient of reduced Melastatin expression with increased dermal depth (3 of 14). Uniform expression of Melastatin mRNA was observed in 49% of primary cutaneous melanomas (18 of 37 cases, including 1 case of in situ melanoma). Melastatin mRNA loss by a portion of the melanoma was identified in 53% of the invasive melanoma samples (19 of 36) and 100% of the melanoma metastases (11 of 11). Primary melanomas without mRNA loss ranged in thickness from 0.17 to 2.75 mm (median, 0.5 mm; mean, 0.73 mm), whereas tumors that showed Melastatin mRNA down-regulation ranged in thickness from 0.28 to 5.75 mm (median, 1.7 mm; mean, 2.13 mm). A focal aggregate or nodule of melanoma cells without detectable signal was the most commonly observed pattern of Melastatin loss (13 of 19 cases), whereas complete loss of Melastatin mRNA expression by all of the dermal melanoma cells was observed in only 4 of the 19 cases. Two invasive melanomas displayed a scattered, nonfocal pattern of Melastatin mRNA loss. Of the 11 melanoma metastases examined, 64% displayed focal Melastatin mRNA loss, and 36% had complete loss of Melastatin mRNA expression. We observed several patterns of Melastatin mRNA expression in primary melanoma that may be distinguished from expression in benign melanocytic nevi. Melastatin mRNA expression appears to correlate with melanocytic tumor progression, melanoma tumor thickness, and the potential for melanoma metastasis. HUM PATHOL 31:1346:1356.     

Expression of interleukin-8 detected by in situ hybridization correlates with worse prognosis in primary cutaneous melanoma

Previous in vitro studies have demonstrated that endogenously produced human interleukin-8 (IL-8) can act as an important growth factor for human melanoma cells in vitro. The present study, has investigated whether IL-8 mRNA expression in primary melanomas may be of prognostic relevance with regard to melanoma progression and metastatic spread. In order to evaluate the clinical significance of IL-8 mRNA expression of melanoma cells in vivo, 59 melanocytic tissue specimens (37 primary melanomas and 22 melanocytic naevi) were studied using a semiquantitative in situ hybridization technique. Significant mRNA expression of IL-8 was found in 59 per cent (22/37) of melanomas. In 19 per cent (7/37) of the malignant melanomas, additional hybridization signals were noted within keratinocytes of the overlying epidermis. In contrast, paralesional normal-appearing epidermis and melanocytes in non-malignant lesions (melanocytic naevi) showed no IL-8 mRNA. Analysis of the relationship between IL-8 expression and clinico-histopathological features showed a significant association between IL-8 mRNA expression and the histological melanoma subtype (IL-8 mRNA: 14/19 in superficial spreading melanoma versus 4/12 in nodular melanoma, p< 0.05). Furthermore, IL-8 expression in primary tumours could be correlated with the patients' clinical course, with time to progression being significantly reduced in primary tumours expressing IL-8 in either the tumour cells or keratinocytes of the overlying epidermis. These results demonstrate for the first time that IL-8 expression, as detected by in situ hybridization in primary tumours, may serve as a significant prognostic factor for tumour progression in human malignant melanoma.     

The status of microRNA-21 expression and its clinical significance in human cutaneous malignant melanoma

Dysregulation of microRNA-21 plays critical roles in tumor initiation and progression. The purpose of this study was to investigate the status of microRNA-21 expression in human cutaneous malignant melanoma and determine its clinical significance. TaqMan^® real-time RT-PCR assay was performed to examine the expression of microRNA-21 in 10 cases of dysplastic nevi, 86 cases of primary cutaneous melanomas, 10 cases of melanoma metastases. The correlation of microRNA-21 expression with clinicopathological factors or prognosis of patients with cutaneous melanoma was statistically analyzed. Additionally, the effects of microRNA-21 expression on growth, apoptosis and chemo- or radiosensitivity of melanoma cells were also investigated by transfection of microRNA-21 inhibitor. We firstly showed that increased levels of microRNA-21 expression were shown from dysplastic nevi to primary cutaneous melanomas to melanoma metastases. Moreover, high miR-21 expression was found to be correlated with Breslow thickness and advanced clinical stage. Patients with high microRNA-21 expression showed shorter 5-year disease-free or overall survival than those with low microRNA-21 expression. Furthermore, multivariate regression analysis showed that the status of microRNA-21 expression was an independent prognostic factor for overall survival of patients. Antisense-mediated microRNA-21 inhibition could significantly suppress growth, increase apoptosis and enhance chemo- or radiosensitivity of human cutaneous melanoma cells by inducing the increased Bax/Bcl-2 ratio. Thus, the status of microRNA-21 might be an independent prognostic factor for patients with cutaneous melanoma, and microRNA-21 has the potential of being a novel molecular target for the treatment of human cutaneous melanoma.     

Prognostic value of tumour thickness in cutaneous malignant melanoma

The relation between survival and histological features in 91 patients with malignant melanoma was studied and the results were analysed by Clayton's method for interpretation of censored survival data. There was a significant correlation between tumour thickness and survival. The risk of dying from malignant melanoma after 10 years of follow up was less than 15% if the primary tumour was less than 1·5 mm thick but more than 80% if the lesion was thicker than 8 mm.     

Treatment and Outcomes of Melanoma in Acral Location in Korean Patients

Purpose

A retrospective study was conducted to review the treatment and outcomes of mainly melanomas in acral location in a single institution in Korea, and to evaluate the prognostic significance of anatomic locations of the tumor.

Materials and Methods

A retrospective review was completed on 40 patients between 2001 and 2006 to obtain pertinent demographic data, tumor data, treatment characteristics, and follow-up data.

Results

Forty melanoma patients were identified and analyzed. Of these, 18 were male and 22 were female patients and the mean age at the time of diagnosis was 55.9 years. Of the tumors, 65% were located on the hands and feet with acral lentiginous melanoma being the most common histological subtype. Univariate analysis for the overall melanoma survival revealed that the thickness of the tumor and the clinical stage have prognostic significances. The most significant factor as analyzed by a multivariate analysis was shown to be the advanced clinical stage. Acral melanomas did not show statistically significant differences in the age at diagnosis, thickness of the tumor, stage, ulceration, and survival rates compared to non-acral melanomas. There was also no significant difference in the survival rate between the patients treated by amputation versus wide local excision in acral melanomas.

Conclusion

In Korean melanoma patients, thickness and advanced stages are significant factors for poorer prognosis. However, the location of melanoma did not have a significant prognostic value. In treating the melanomas in acral location, local wide excisions resulted in a similar prognosis compared to amputations.     

凝血酶受体-1在肺癌组织中的表达及其与转移的关系

目的研究凝血酶受体（PAR）-1在肺癌组织中的表达及其与肺癌侵袭、转移的关系。方法免疫组织化学sP法、形态学计量及逆转录-聚合酶链反应（RT-PCR）检测肺癌原发灶和转移灶组织（36例液氮冻存肺癌组织、80例石蜡包埋肺癌组织）中PAR-1的表达。结果具有侵袭和转移部位的癌巢、脉管内癌栓、癌周围的肺泡上皮不典型增生灶及支气管腺导管上皮不典型增生灶均呈现较强的阳性反应。肺癌组织PAR-1蛋白表达阳性率为73．8％（59／80例）;转移组85．7％（48／56）与非转移组45,8％（11／24）之间差异有统计学意义（P〈0．05）。转移和非转移（P〈0．05）、原发灶和转移灶（P〈0．05）、肿瘤组织和肺组织（P〈0．01）各组之间PAR．1蛋白含量差异有统计学意义;而肿瘤大小、组织学类型和组织学分化各组间差异无统计学意义（P〉0．05）。肺癌组织PAR．1mRNA表达阳性率63．9％（23／36例）;转移组78．3％（18／23例）与非转移组38．5％（5／13）之间差异有统计学意义（P〈0．05）。结论PAR-1过度表达与肺癌的转移表型、组织发生及恶性表型有关;PAR-1可能是肺癌转移过程中发挥重要作用的因素之一。     

Poly(adenosine diphosphate-ribose) polymerase 1 expression in malignant melanomas from photoexposed areas of the head and neck region

The family of the poly(adenosine diphosphate-ribose) polymerase (PARP) proteins is directly involved in genomic stability, DNA repair, and apoptosis by DNA damage. In this study, we evaluated the role of PARP-1 in melanoma and its prognostic importance. We studied by immunohistochemistry and Western blot analysis PARP-1 expression in a selected series of 80 primary melanoma of the head and neck region. The results were correlated with tumor thickness and patient's outcome. A follow-up of at least 3 years was available. Fifteen cases of benign melanocytic nevi were used as controls. Normal melanocytes showed only scattered, focal nuclear positivity and were considered as negative for PARP-1 expression by immunohistochemistry (score, 0). Thirty cases of melanoma (37.5%) showed nuclear expression of PARP-1 in both radial and vertical growth phases. Western blot analysis showed the presence of a high signal for full-length PARP-1 only in the cases with high immunohistochemical (nuclear) expression of protein (score, ++/+++) in both radial and vertical growth phase. A significant correlation was present between PARP-1 expression in vertical growth phase and the thickness of tumor lesion (P = .014); all but one tumor measuring less than 0.75 mm showed no or low PARP-1 expression. No correlation was found between PARP-1 expression in radial growth phase and tumor thickness (P = .38, data not shown). These data suggest that PARP-1 overexpression is a potential novel molecular marker of aggressive cutaneous malignant melanoma and a direct correlation between PARP-1-mediated inhibition of the apoptosis and biologic behavior of cutaneous malignant melanoma.     

CCL27-CCR10 and CXCL12-CXCR4 chemokine ligand-receptor mRNA expression ratio: new predictive factors of tumor progression in cutaneous malignant melanoma

CXCR4, CCR7 and CCR10 chemokine receptors are known to be involved in melanoma metastasis. Our goal was to compare the relative intratumoral mRNA expression of these receptors with that of their corresponding chemokine ligands, CXCL12, CCL19, CCL21, and CCL27 across the full spectrum of human melanoma progression: thin and thick primary melanomas, as well as "in transit", lymph node, and distant metastases. Expression was quantified by real-time RT-PCR in 103 melanoma samples: 51 primary tumors and 52 metastases. Particular emphasis was focused on chemokine ligand-receptor expression ratios. Immunohistochemistry was performed to identify the cell types expressing these molecules. CXCL12-CXCR4 and CCL27-CCR10 ratios were higher in thin than in thick primary melanomas, and all four chemokine-receptor ratios were higher in primary tumors than in melanoma metastases. CCL27-CCR10 and CXCL12-CXCR4 expression ratios in primary tumors were inversely associated with the development of distant metastases, and improved the predictive value of tumor thickness for distant metastasis, which is important since chemokine ligand-receptor ratios are not affected by the endogenous gene employed for normalizing mRNA expression. Both receptor and ligand immunolabeling were detected in neoplastic cells suggesting autocrine mechanisms. Our results support the concept that low CCL27/CCR10 and CXCL12/CXCR4 intratumoral mRNA ratios are associated with melanoma progression, and in combination with Breslow thickness, are the best predictive factors for the development of distant metastases in primary cutaneous melanoma.     

Partial regression in thin primary cutaneous malignant melanomas clinical stage I

Summary 486 patients with primary cutaneous malignant melanoma clinical stage I were examined in order to evaluate the prognostic importance of partial regression in thin lesions. All the melanomas measured 1 mm or less in maximal thickness. The study showed that past regression with fibrotic scar tissue adversely affected survival in patients with thin melanomas. The 10 year survival was 95% for patients without regression in contrast to 79% for patients with past regression. It was, furthermore, demonstrated that active regression without fibrotic scar tissue did not influence survival significantly.The wider and the thicker the fibrotic area, the poorer the survival. Although the prognostic importance of this finding was not statistically significant, we suggest that the horizontal width of the fibrotic area in particular may be a valuable prognostic guide in thin melanomas with past regression.     

Review: polymerase chain reaction detection of micrometastases and circulating tumor cells: application to melanoma, prostate, and thyroid carcinomas.

The main strategy used for the detection of circulating tumor cells (CTC) and micrometastases in solid tumors is the polymerase chain reaction (PCR) amplification of tissue specific messenger RNA present in the tumor cells. PCR was more sensitive than conventional techniques, allowing the identification of one tumor cell diluted into 1 mL of blood. PCR was shown to be specific in most studies related to the detection of CTC and marrow micrometastases in melanoma and prostate carcinoma (PC). PCR positivity for thyroid markers was reported in the blood of control subjects. Large variations in the PCR positivity rates and the prognostic value of these assays have been encountered in PC and melanoma. There was a correlation between PCR and stage in some but not all the studies. Despite these discrepancies, many investigators have shown PCR to be predictive of outcome in PC and especially in melanoma. PCR in blood and bone marrow was an independent predictor of overall and disease-free survival in melanoma patients rendered surgically free of disease. These tests may help better stratify patients for radical surgeries and adjuvant therapy. Large prospective and interlaboratory studies are needed to confirm the accuracy and prognostic value of these assays.