Eves Paula, Katerinaki Efthymia, Simpson Claire, Layton Christopher, Dawson Rebecca, Evans Gareth, Mac Neil Sheila
Skin Biology Group, Division of Clinical Sciences (North), University of Sheffield, Northern General Hospital, Sheffield, UK.
Clin Exp Metastasis. 2003;20(8):685-700. doi: 10.1023/b:clin.0000006824.41376.b0.
Melanoma invasion is a complex multi stage process involving changes to the cell/extracellular matrix (ECM) and cell/cell interactions. We have previously shown using an in vitro model of reconstructed human skin (consisting of human dermis with a basement membrane [BM] and populated with human skin cells) that some melanoma cells (HBL cell line) invade more actively in the presence of adjacent normal skin cells. The aim of the present study was to further investigate the relationship between melanoma cells, skin cells and ECM proteins during melanoma cell invasion through reconstructed skin, extending this to a study of three melanoma cell lines. We also examined whether such cell/cell induced invasion is due to increased expression and activation of matrix-metalloproteinase-2 (MMP-2) and MMP-9, or due to increases in general protease activity for keratinocytes, fibroblasts or melanoma lines. Addition of skin cells dramatically altered the invasive behaviour of the three metastatic melanoma cell lines (HBL, C8161 and A375SM) used; they increased the invasive ability of HBLs which were unable to invade on their own; they potentiated the invasion of C8161 cells which were invasive in their own right, but reduced the invasion of A375-SM cells which were aggressive invaders in the absence of skin cells. Latent forms of MMP-2, and MMP-9, were clearly expressed by the normal skin cells whereas all three melanoma lines weakly expressed these proteases. Fibroblast and keratinocyte MMPs were activated specifically by culture on type I collagen and on dermis which retained an intact basement membrane. These findings demonstrate that while there is an active communication between melanoma cells and adjacent skin cells, the invasive process is dictated by the melanoma cells and not the skin cells. However, activation of skin cell derived MMPs may play an important role in facilitating invasion by particular melanoma phenotypes.
黑色素瘤侵袭是一个复杂的多阶段过程，涉及细胞/细胞外基质（ECM）和细胞/细胞相互作用的改变。我们之前使用重建人皮肤的体外模型（由带有基底膜[BM]的人真皮组成，并接种人皮肤细胞）表明，一些黑色素瘤细胞（HBL细胞系）在相邻正常皮肤细胞存在的情况下侵袭更为活跃。本研究的目的是进一步研究黑色素瘤细胞侵袭重建皮肤过程中黑色素瘤细胞、皮肤细胞和ECM蛋白之间的关系，并将其扩展到对三种黑色素瘤细胞系的研究。我们还研究了这种细胞/细胞诱导的侵袭是否是由于基质金属蛋白酶-2（MMP-2）和MMP-9的表达和激活增加，或者是由于角质形成细胞、成纤维细胞或黑色素瘤细胞系的一般蛋白酶活性增加。添加皮肤细胞显著改变了所使用的三种转移性黑色素瘤细胞系（HBL、C8161和A375SM）的侵袭行为；它们增加了原本无法自行侵袭的HBL细胞的侵袭能力；增强了本身具有侵袭性的C8161细胞的侵袭能力，但降低了在没有皮肤细胞时具有侵袭性的A375-SM细胞的侵袭能力。正常皮肤细胞明显表达MMP-2和MMP-9的潜伏形式，而所有三种黑色素瘤细胞系均弱表达这些蛋白酶。成纤维细胞和角质形成细胞的MMPs在I型胶原和保留完整基底膜的真皮上培养时被特异性激活。这些发现表明，虽然黑色素瘤细胞与相邻皮肤细胞之间存在活跃的通讯，但侵袭过程由黑色素瘤细胞而非皮肤细胞决定。然而，皮肤细胞衍生的MMPs的激活可能在促进特定黑色素瘤表型的侵袭中起重要作用。
