Lymph node micrometastases of cutaneous melanoma: Increased sensitivity of molecular diagnosis in comparison to immunohistochemistry

Blaheta, Hans-Juergen; Schittek, Birgit; Breuninger, Helmut; Maczey, Evelyn; Kroeber, Stefan; Sotlar, Karl; Ellwanger, Ulf; Thelen, Marcel H.; Rassner, G.; Bültmann, B.; Garbe, Claus

doi:10.1002/(sici)1097-0215(19980821)79:4<318::aid-ijc3>3.0.co;2-y

Cited by 75 publications

(42 citation statements)

References 21 publications

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“…These results are conflicting with the recent study by Blaheta et al (1998) who found no positive results in lymph nodes of 40 non-melanoma patients. However, it is to be noted that in this study, together with the nested RT-PCR an immunohistochemical assay, is performed and eventual positive results are judged to be positive only when confirmed by the detection of tyrosinasepositive cells by means of immunohistochemistry.…”

Section: Discussioncontrasting

confidence: 80%

“…Although clinical parameters such as lymph node involvement and thickness of the tumour are helpful, other diagnostic tools are urgently needed to allow for optimal prediction of the clinical course and choice of treatment. For this purpose efforts have been made to detect micrometastases in blood (Brossart et al, 1995;Foss et al, 1995;Hoon et al, 1995;Buzaid and Balch 1996;Mellado et al, 1996;Gläser et al, 1997;Jung et al, 1997;Reinhold et al, 1997;Farthmann et al, 1998;Ghossein et al, 1998;Curry et al, 1998;De Vries et al, 1999;Palmieri et al, 1999) and in the regional lymph nodes by immunohistochemistry or reverse transcriptase polymerase chain reaction (RT-PCR) (Battyani et al, 1993;Wang et al, 1994;Rankin, 1996;Van der Velde-Zimmermann et al, 1996;Blaheta et al, 1998;Goydos et al, 1998;Hatta et al, 1998;Bieligk et al, 1999).…”

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confidence: 99%

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Limitations of the nested reverse transcriptase polymerase chain reaction on tyrosinase for the detection of malignant melanoma micrometastases in lymph nodes

Calogero

Timmer‐Bosscha

Koops³

et al. 2000

Br J Cancer

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SummaryThe specificity and sensitivity of the nested reverse transcriptase polymerase chain reaction (RT-PCR) on tyrosinase was studied, for the detection of micrometastases of malignant melanoma. The specificity was assessed in the blood of six healthy donors, four patients with non-melanoma cancers of which one patient was treated with granulocyte-colony stimulating factor. Lymph nodes of nine patients without malignant melanoma were tested and four cell lines of various other tumours. Six of the nine non-melanoma lymph nodes were positive in this assay. The sensitivity was tested in a spike experiment in vitro, using a melanoma cell line. The detection limit was ten melanoma cells per 10 7 peripheral blood lymphocytes.

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Section: Discussioncontrasting

confidence: 80%

mentioning

confidence: 99%

Limitations of the nested reverse transcriptase polymerase chain reaction on tyrosinase for the detection of malignant melanoma micrometastases in lymph nodes

Calogero

Timmer‐Bosscha

Koops³

et al. 2000

Br J Cancer

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“…Results from these studies are promising, as the technique was found to be sensitive and specific, and results are associated with clinical stage and prognosis. [11][12][13][14] Furthermore, the presence of CMCs has been found to be a useful predictor of relapse and death. 15 In patients with cutaneous melanoma, Tyrosinase and MART-1/Melan A (melanocyte-specific genes) have been successfully used to identify CMCs in peripheral blood samples.…”

Section: Introductionmentioning

confidence: 99%

Identification of circulating malignant cells and its correlation with prognostic factors and treatment in uveal melanoma. A prospective longitudinal study

Callejo

Antecka

Blanco

et al. 2006

Eye

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Purpose Most uveal melanoma patients (UMP) do not show evidence of metastases upon diagnosis. However, despite local tumour control, B50% of them will develop metastases. These findings suggest that malignant cells may have already disseminated by the time of initial diagnosis. The purpose of the study was to detect circulating malignant cells (CMCs) in UMP and to correlate them with prognostic factors and therapy. Methods Nested reverse transcriptasepolymerase chain reaction (RT-PCR) was used to detect CMCs. In each UMP, blood was collected every 3 months. In each visit, 20 RT-PCR tests were performed. The date of diagnosis, largest tumour dimension, type, and date of treatment were obtained. Results A total of 30 UMP were enrolled. Five patients were enrolled at the time of diagnosis and 25 patients between 1 and 17 years following diagnosis. No UMP showed clinical evidence of metastasis. A total of 136 visits were registered, 1360 samples collected, and 2720 RT-PCRs performed. CMCs were identified in 29 patients in 119 visits (87.5%). However, in each visit, a low number of positive tests were recorded. CMCs were found in newly diagnosed, irradiated, enucleated, and observed patients regardless of tumour size and time period following treatment. Conclusions Uveal melanoma (UM) is not a localized ocular disease. CMCs were recorded at initial diagnosis confirming the early metastatic nature of UM. CMCs were present following treatment, including enucleation, demonstrating that CMCs are capable of disseminating and surviving, possibly as micrometastasis, which would contribute to the pool of CMCs at a later stage. Systemic therapy should be evaluated.

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“…PCR-based assays are extremely sensitive and are less vulnerable to intranodal tumour cell distribution when compared to IHC. 14,25,26 It is of course critical to have a representative amount of the SLN available for analysis which outweighs that of multiple sectioning. Ideally, the entire lymph node or half of the lymph node cut along its longitudinal axis should be used for PCR-based assays.…”

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confidence: 99%

HPV16‐E6 mRNA is superior to cytokeratin 19 mRNA as a molecular marker for the detection of disseminated tumour cells in sentinel lymph nodes of patients with cervical cancer by quantitative reverse‐transcription PCR

Häfner

Gajda

Altgassen

et al. 2007

Intl Journal of Cancer

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About 10-15% of patients with cervical cancer suffer from recurrence despite histologically negative lymph nodes (pN0). Occult micrometastases or small tumour cell clusters may contribute to disease outcome. The aim of this study was to compare at the RNA level 2 known tumour-associated genes, HPV16-E6 and cytokeratin 19 (CK19), as molecular markers for the detection of disseminated tumour cells. Real-time reverse transcription PCR technology was used to quantify gene expression in histologically positive and negative sentinel lymph nodes (SLN) from 70 patients with cervical cancer. Lymph nodes from noncancer patients were used as controls. Calculated copy numbers were normalised to the geometric average of the most stable housekeeping genes. We observed a good correlation (R 5 0.915) between the expression of both markers in SLN with histologically confirmed metastases. However, marker gene expression differed considerably in histologically negative nodes: CK19 transcripts were detected in 90 of 112 SLN (80.4%), whereas only 38 nodes (33.9%) were positive for HPV16 E6 mRNA. In particular, 62 of 74 SLN, which were negative by histology, and HPV16 E6 mRNA expressed CK19 mRNA. Moreover, 8 of 10 lymph nodes from noncancer patients expressed CK19 mRNA. Systematic errors due to RNA degradation or incomplete cDNA could be ruled out. It is concluded that HPV16 E6 mRNA is more specific and more sensitive for the detection of tumour cells in SLN than CK19 mRNA. The specificity of CK19 is limited because of low level expression in uninvolved pelvic lymph nodes. ' 2007 Wiley-Liss, Inc.Key words: cervical cancer; disseminated tumour cells; molecular markers; HPV16-E16; cytokeratin 19; quantitative RT-PCR Cervical cancer is the second most common cancer among women world wide, accounting for 493,000 new cases in 2002. 1 The incidence rates vary considerably between different geographic regions, which reflect in part the lack of screening programs in developing countries. Despite the advancement in early diagnosis, improved surgical techniques and adjuvant therapies, approximately 40% of cervical carcinoma patients in countries with a high standard of health care die from the disease. 1 The majority of these deaths is due to metastases. 2 Different parameters such as lymph node metastases, tumour grade, lymph-vascular space involvement (LVSI) and surgical margins are prognostic factors for cervical cancer, with lymph node status as the most important one. 3,4 Patients with histologically uninvolved lymph nodes (pN0) have a better prognosis than women with histologically confirmed lymph node metastasis (pN1) showing 5 year survival rates of 88-93% and 40-63%, respectively. 5-7 Nevertheless 15% of FIGO stage IB patients with pathologically staged negative lymph nodes (pN0) suffer from recurrence. 8 The significance of micrometastases as a predictive factor for recurrent disease is the focus of several recent and ongoing studies. Extensive serial sectioning of lymph nodes that were judged to be free of disease by routine diagnos...

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Lymph node micrometastases of cutaneous melanoma: Increased sensitivity of molecular diagnosis in comparison to immunohistochemistry

Cited by 75 publications

References 21 publications

Limitations of the nested reverse transcriptase polymerase chain reaction on tyrosinase for the detection of malignant melanoma micrometastases in lymph nodes

Limitations of the nested reverse transcriptase polymerase chain reaction on tyrosinase for the detection of malignant melanoma micrometastases in lymph nodes

Identification of circulating malignant cells and its correlation with prognostic factors and treatment in uveal melanoma. A prospective longitudinal study

HPV16‐E6 mRNA is superior to cytokeratin 19 mRNA as a molecular marker for the detection of disseminated tumour cells in sentinel lymph nodes of patients with cervical cancer by quantitative reverse‐transcription PCR

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