Abstract
CD63, which belongs to the tetraspanin membrane proteins, has been proposed to play an important role in inhibiting melanoma metastasis. To determine whether reduction of CD63 expression, which frequently occurs in the malignant progression of human melanoma, is responsible for metastasis promotion, we transfected the antisense CD63 cDNA into MelJuso melanoma cells having endogenous CD63 expression. The antisense CD63 transfectant clones showing decreased CD63 expression displayed increased cell motility, matrix- degrading activity, and invasiveness in vitro when compared with the control transfectant cells. The antisense CD63 cDNA-transfected cells also exhibited altered adhesiveness to extracellular matrix. The results suggest that reduced CD63 expression contributes to the invasive and metastatic ability of human melanoma cells.
Highlights
Tetraspanins, or transmembrane 4 superfamily (TM4SF) molecules, are characterized by the existence of four highly conserved transmembrane domains and some members of this family such as CD9, CD63, CD81, CD82, and CD151 are ubiquitously expressed whereas others are highly restricted to immune cells (Maecker et al, 1997; Boucheix and Rubinstein, 2001; Yanez-Mo et al, 2001, for reviews)
CD63 tetraspanin protein, which has been identified as a melanoma-associated antigen ME491, is another candidate that might play a role in cancer metastasis
C8161 and MelJuso human melanoma cell lines were cultured in DMEM/F-12 medium supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin in 5% CO2 at 37oC
Summary
Tetraspanins, or transmembrane 4 superfamily (TM4SF) molecules, are characterized by the existence of four highly conserved transmembrane domains and some members of this family such as CD9, CD63, CD81, CD82, and CD151 are ubiquitously expressed whereas others are highly restricted to immune cells (Maecker et al, 1997; Boucheix and Rubinstein, 2001; Yanez-Mo et al, 2001, for reviews). The tetraspanin molecules have been implicated in diverse biological phenomena, including cell proliferation, activation, adhesion, migration, differentiation, and development. Some of these molecules have been implicated in cancer metastasis: CD9 expression is inversely correlated with the appearance of metastases in melanomas, breast, lung, and colon cancers (Si and Hersey, 1993; Miyake et al, 1995; Adachi et al, 1998; Mori et al, 1998). We transfected CD63 cDNA in antisense direction into a human melanoma cell line having endogenous CD63 expression, MelJuso, and examined the effect of reduced CD63 expression on the metastatic phenotype of MelJuso melanoma cells in vitro
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