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Loss of microRNA-200a and c, and microRNA-203 expression at the invasive front of primary cutaneous melanoma is associated with increased thickness and disease progression

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Abstract

Loss of E-cadherin expression in melanoma correlates with increased tumor thickness and reduced disease-free survival. The molecular mechanisms underpinning its differential expression in melanoma tissue remain elusive. MicroRNAs (miRNAs) have been implicated in tumor progression and regulation of E-cadherin expression. Here, we demonstrate a significant correlation between tumor thickness and loss of expression of miR-200a, miR-200c, and miR-203 in a series of 23 frozen primary melanomas, where it was confirmed in two subsequent validation series (series 1: six nevi, 15 primary melanomas, and 16 metastases; series 2: 11 matched pairs of primary melanomas and metastases). Decreased levels of miR-200a, miR-200c, and miR-203 correlated with increasing thickness in the combined validation series (P = 0.024, 0.033, and 0.031, respectively). In addition, progressive loss of miR-200a expression with disease progression was observed in series 1 (P < 0.001) and in series 2 (P = 0.029). MiR-200 in situ hybridization and E-cadherin immunohistochemistry demonstrated reduced expression of both at the deep invasive margin of the tumor. Furthermore, a functional validation study using an anti-miR200 strategy demonstrated that loss of miR-200 expression in melanoma cell lines reduced E-cadherin expression. Collectively, our data point towards an important role for miR-200 and miR203 expression in regulating E-cadherin during melanoma progression.

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Authors and Affiliations

  1. Lady Davis Institute for Medical Research, Jewish General Hospital, LDI-E447, 3755 Cote Ste-Catherine, Montreal, QC, Canada, H3T 1E2

    Léon C. van Kempen & Alan Spatz

  2. Department of Pathology, McGill University, Jewish General Hospital, Montreal, Canada

    Léon C. van Kempen & Alan Spatz

  3. Department of Oncology, McGill University, Jewish General Hospital, Montreal, Canada

    Alan Spatz

  4. Department of Pathology, GROW-School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands

    Karin van den Hurk & Véronique Winnepenninckx

  5. Division of Functional Genomics, Gustave-Roussy Institute, Villejuif, France

    Vladimir Lazar

  6. Division of Biostatistics and Epidemiology, Gustave-Roussy Institute, Villejuif, France

    Stefan Michiels

  7. Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium

    Stefan Michiels

  8. Division of Surgical Oncology, University Hospitals, Katholieke Universiteit Leuven, Leuven, Belgium

    Marguerite Stas

  9. Laboratory of Morphology and Molecular Pathology, University Hospitals, Katholieke Universiteit Leuven, Leuven, Belgium

    Joost J. van den Oord

  10. EORTC Melanoma Group, Brussels, Belgium

    Léon C. van Kempen, Marguerite Stas, Alan Spatz & Joost J. van den Oord

  11. Department of Pathology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

    Léon C. van Kempen

Authors
  1. Léon C. van Kempen
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  2. Karin van den Hurk
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  3. Vladimir Lazar
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  8. Joost J. van den Oord
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Corresponding author

Correspondence to Léon C. van Kempen.

Electronic supplementary material

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Figure S1

In situ detection of miR-200 and E-cadherin in a desmoplastic melanoma. Consecutive tissue sections of a desmoplastic melanoma (A, H&E staining) were subjected to Ecadherin immunohistochemistry (B,C) and miR-200c in situ hybridization (D,E). E-cadherin and miR-200c expression was not observed in the tumor (B and D, respectively), but the overlying epidermis was strongly positive for membranous E-cadherin (C, brown staining) and cytoplasmic miR-200c (E, blue staining; arrows indicate a few of many positive cells) expression (PDF 246 kb)

Table S1

List of 23 patients with primary melanomas and their pertinent clinical and histological features, used in the discovery phase of the study (PDF 44 kb)

Table S2A

List of 15 patients with primary melanomas and their pertinent clinical and histological features, used in the first validation phase of the study (PDF 40 kb)

Table S2B

List of 16 patients with melanomas metastases and 6 nevi used in the first validation phase of the study (PDF 33 kb)

Table S3

Pertinent clinical data and Ct values of the set of 11 paired primary-metastatic melanomas used in the second validation study (PDF 34 kb)

Table S4

Spread sheets of all miRNAs analysed, raw Ct values and calculated values (PDF 966 kb)

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van Kempen, L.C., van den Hurk, K., Lazar, V. et al. Loss of microRNA-200a and c, and microRNA-203 expression at the invasive front of primary cutaneous melanoma is associated with increased thickness and disease progression. Virchows Arch 461, 441–448 (2012). https://doi.org/10.1007/s00428-012-1309-9

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  • DOI: https://doi.org/10.1007/s00428-012-1309-9

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