CD44v3 levels in primary cutaneous melanoma are predictive of prognosis: Assessment by the use of tissue microarray

Wayne State University, Detroit, Michigan, United States
International Journal of Cancer (Impact Factor: 5.09). 03/2006; 118(6):1460-4. DOI: 10.1002/ijc.21504
Source: PubMed


Despite the use of sentinel node biopsy techniques, the search continues for other strategies to improve the accuracy of estimating prognosis in melanoma patients. Various biomarkers have previously been studied for use in this role, but none has yet achieved acceptance in routine practice. We have applied the novel technology of tissue microarray for the high throughput screening of a cohort of 120 primary cutaneous melanoma specimens for expression of the transmembrane glycoprotein CD44, splice variant 3 (v3), which has previously been implicated in tumor progression. A highly significant correlation between CD44v3 expression and Breslow thickness, Clark's level and patient age was demonstrated (Spearman correlation p < 0.001). Regarding clinical outcome, CD44v3 expression was shown to be significantly associated with better outcome (chi(2) = 7.2219, p = 0.0072). Furthermore, subgroup analysis revealed a sequentially improved survival probability associated with the intensity of CD44v3 staining (chi(2) = 12.5162, p = 0.0058). Analysis in a Cox multivariate model, however, did not show CD44v3 to be independently predictive of prognosis. The implications of these findings are considered, and the use of CD44v3 as a potential prognostic marker or a target for therapeutic manipulation are discussed.

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Available from: George D Wilson, Sep 25, 2014
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    • "Similar discrepancies emerge in the changing patterns of CD44v expression in human melanoma. Dome et al. demonstrated the up-regulation of v3 results in a higher visceral metastatic potential [21] while Pacifico et al. showed that CD44v3 expression is associated significantly with a better outcome [22]. Again v10 [23] and v6 [24] may be involved in metastasis formation, or not [25]. "
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    ABSTRACT: The role of CD44 in the progression of human melanoma has mostly been characterised by qualitative changes in expression of its individual variable exons. These exons however, may be expressed to form a number of molecules, the alternative splice variants of CD44, which may be structurally and functionally different. Using real-time PCR measurements with variable exon specific primers we have determined that all are expressed in human melanoma. To permit comparison between different tumours we identified a stable CD44 variable exon (CD44v) expression pattern, or CD44 ‘fingerprint’. This was found to remain unchanged in melanoma cell lines cultured in different matrix environments. To evaluate evolution of this fingerprint during tumour progression we established a scid mouse model, in which the pure expression pattern of metastatic primary tumours, circulating cells and metastases, non-metastatic primary tumours and lung colonies could be studied. Our analyses demonstrated, that although the melanoma CD44 fingerprint is qualitatively stable, quantitative changes are observed suggesting a possible role in tumour progression.
    PLoS ONE 01/2013; 8(1):e53883. DOI:10.1371/journal.pone.0053883 · 3.23 Impact Factor
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    • "The role of CD44v3 in cancer is tumor type dependent; however in many malignancies studied, its expression is associated with a better prognosis compared to tumors that lack CD44v3 expression. In melanoma, CD44v3 expression is associated with other key prognostic factors resulting in a 75% 5 year survival in CD44v3 positive tumors vs. 45% in negative cases, p = 0.0072 [24]. Similarly, CD44v3 is present in the majority of benign uterine fibroids but is uniformly absent in uterine sarcomas [25]. "
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    BMC Cancer 05/2012; 12(1):176. DOI:10.1186/1471-2407-12-176 · 3.36 Impact Factor
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    ABSTRACT: Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) is a versatile tool for precise quantification of gene expression. Formalin-fixed and paraffin-embedded (FFPE) tissue is well suited for qRT-PCR, if RNA extraction is optimized and small amplicon sizes are used. However, little is known whether individual assays may show variable sensitivity to fixation. This is of great importance, if a direct comparison of different transcripts is performed within the same sample, such as for mRNA splice variants. We established a cell culture model to test for and quantify differences in performance of individual qRT-PCR assays on FFPE as compared with fresh material, using TaqMan methodology. RNA was isolated from 7 different cell lines either directly or after preparation of a FFPE cell block. RNA from both sources was reverse transcribed and gene expression quantified using 13 different TaqMan assays. All assays allowed highly reproducible target quantification, using both fresh and FFPE-derived cDNA. The 13 assays showed an average Ct difference of 3.2 between fresh and FFPE cells, if identical amounts of cDNA were used as template. However, the Ct shifts varied from 1.8 to 5.1 for individual assays, indicating variable resistance to fixation. These Ct shift differences were statistically highly significant in 27/78 (35%) of all possible combinations of assays. Because the Ct shift remained constant for each assay, they could be used for calculation of correction factors which rendered FFPE-derived expression data highly comparable to those obtained from fresh material, and as a consequence among each other. Thus, a standardized assessment of qRT-PCR assay efficiencies in FFPE allows for precise intraindividual comparison of mRNA species, such as splice variants with different biologic functions, in archival tissues.
    Diagnostic Molecular Pathology 10/2006; 15(3):149-56. DOI:10.1097/01.pdm.0000213450.99655.54 · 1.86 Impact Factor
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