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. 2001 Jun;85(6):732–738. doi: 10.1136/bjo.85.6.732

A comparison of ocular melanocyte and uveal melanoma cell invasion and the implication of α1β1, α4β1 and α6β1 integrins

S Elshaw 1, K Sisley 1, N Cross 1, A Murray 1, S MacNeil 1, M Wagner 1, C Nichols 1, I Rennie 1
PMCID: PMC1723995  PMID: 11371496

Abstract

BACKGROUND/AIMS—Posterior uveal melanoma is the most common intraocular tumour in adults, responsible for the death of approximately 35% of patients. Hepatic metastases are most frequent, and once diagnosed survival is usually less than 1 year. The β1 family of integrins, αvβ3 and MMP-2 and MMP-9 have been implicated in the metastasis of several types of tumour. To study their involvement in uveal melanoma we analysed the expression of the β1 integrins, αvβ3, MMP-2, and MMP-9 in 10 primary posterior uveal melanomas, and correlated expression with invasive potential in vitro. Comparable studies were undertaken on cultures of melanocytes.
METHODS—Expression of integrins was studied by immunohistochemistry, secretion of MMP-2 and MMP-9 by zymography, and the invasive potential was assessed using a transwell model.
RESULTS—MMP-2 was secreted by all uveal melanomas and seven of 10 secreted MMP-9. Among uveal melanoma, invasion levels of 4-25% were observed and the major integrins expressed were α1β1, α2β1, α3β1, α5β1, and avβ3. Melanocytes did not express α1β1, α4β1, and α6β1.
CONCLUSION—The laminin binding α6β1 integrin was not expressed by either melanocytes or tumours with spindle morphology, which are considered to have a better prognosis. It is possible that expression of the α6β1 integrin may prove useful as a prognostic indicator.



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Figure 1  .

Figure 1  

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Percentage invasion of 10 cultures of primary posterior uveal melanoma as determined by invasion assay. All cultures were tested within the first five passages. Values shown are means (SEM) for three combined assays.

Figure 2  .

Figure 2  

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Gelatin zymogram showing examples of MMP-2 (72 kD) and MMP-9 (92 kD) secretion by cultured posterior uveal melanoma cells. A molecular weight marker was run on each gel (not shown). Mel 218 (lane 1) also secreted the active forms of both enzymes. Mel 222 (lane 2).

Figure 3  .

Figure 3  

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Photomicrographs showing examples of immunohistochemical staining for integrin expression in cultures of posterior uveal melanoma and melanocytes. (a) Negative staining for α1β1 in melanocytes (×10) (scale bar 100 µm). (b) Homogeneous staining for α3β1 in melanocytes (×40) (scale bar 400 µm). (c) Positive heterogeneous staining of x% of cells for α1β1 in melanoma cells with epithelioid morphology (×20) (scale bar 200 µm). (d) Homogeneous staining for αvβ3 in melanoma cells with epithelioid morphology (×40) (scale bar 400 µm). (e) Homogeneous staining for α6β1 in melanoma cells with epithelioid morphology (×40) (scale bar 400 µm). (f) Negative staining for α6β1 in melanoma cells with spindle-like morphology (×40) (scale bar 400 µm).

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