Human Melanoma Cell Lines Deficient in GD3 Ganglioside Expression Exhibit Altered Growth and Tumorigenic Characteristics

Abstract

We have selected GD3-deficient human melanoma cell lines, in order to investigate the function of GD3 ganglioside. This was done by treating SK-MEL-28 cells with anti-GD3 antibody (R24) and rabbit complement and subsequent subcloning of the surviving cells, resulting in the derivation of two cell lines deficient in the cell surface expression of GD3. Neither cell line (designated SK-MEL-28-N1 and SK-MEL-28-N2) had detectable cell surface expression of GD3 as analyzed with monoclonal antibody R24, and no GD3 was detectable in either cell line by glycolipid isolation, thin-layer chromatography, or resorcinol-HC1 spray, but thin-layer chromatography immunostaining with monoclonal antibody R24 showed the presence of low amounts of GD3 in both N1 and N2 (1/40 of the amount in the parent cell line in N1 and 1/500 in N2). In SK-MEL-28-N1, the residual GD3 was shown by immunofluorescence assays on permeabilized cells to be present in discrete intracellular organelles, suggesting that these cells have a defect in the transport of GD3 as well as in its synthesis. Both SK-MEL-28-N1 and -N2 had an increase in detectable GM3 expression. The mutant cell lines had altered cell morphology in comparison to the parent cell line and both had slower growth rates in vitro and lower tumorgenicity in nu/nu mice. These results indicate that GD3 ganglioside plays an important role in proliferation and growth of melanoma cells.

Ganglioside GD3 promotes cell growth and invasion through p130Cas and paxillin in malignant melanoma cells

“…The facts that GD3 is highly expressed in melanomas in comparison with melanocytes, is exclusively expressed in the early developmental stage of the fetal brain (10), and is highly expressed in activated T lymphocytes and in T cell malignant tumor cells (12)(13)(14)(15) suggest that GD3 is involved in tumor phenotypes, such as enhanced proliferation and frequent metastasis. There have been a number of reports indicating that GD3 is associated with cell adhesion to the extracellular matrix (20), increased cell growth (17), and invasion activity (21). In fact, anti-GD3 antibodies appeared to suppress cultured melanoma growth (16,22), and GD3-lacking mutants of SK-MEL-28 showed markedly reduced cell proliferation (17), supporting its role in cell growth.…”

“…There have been a number of reports indicating that GD3 is associated with cell adhesion to the extracellular matrix (20), increased cell growth (17), and invasion activity (21). In fact, anti-GD3 antibodies appeared to suppress cultured melanoma growth (16,22), and GD3-lacking mutants of SK-MEL-28 showed markedly reduced cell proliferation (17), supporting its role in cell growth. Suppression of GD3 expression with antisense GD3 synthase cDNA also resulted in the reduction of tumor cell phenotypes such as growth, migration, metastasis, and angiogenesis in F11 cells (23,24).…”

“…However, molecular mechanisms for the enhancement of malignant properties by GD3 have never been demonstrated. In this study, we established a set of melanoma cell lines with or without GD3 expression by using a GD3-deficient mutant of SK-MEL-28 that was generated by the treatment of the parent cell with anti-GD3 mAb and complement (17). Reexpression of GD3 with GD3 synthase cDNA provided cell lines that enabled us to identify the phenotypic changes due to GD3 expression in the transfectant cells and also to investigate the molecular mechanisms for the enhanced malignant properties correlated with GD3 expression.…”

“…GD3 is also detected in the T cell malignant leukocytes, such as acute lymphoblastic leukemia cells (11)(12)(13), adult T cell leukemia cells (14), and activated normal human T lymphocytes (5,15). However, no clear function of GD3 in the malignant tumor cells has been demonstrated, although anti-GD3 antibody was reported to suppress the cell growth of cultured melanoma cells (16), and a GD3-negative cell isolate, SK-MEL-28-N1, was found to have slower growth and tumorigenic properties than the parent cell line (17).…”

An Anti‐GD2 Monoclonal Antibody Enhances Apoptotic Effects of Anti‐cancer Drugs against Small Cell Lung Cancer Cells via JNK (c‐Jun Terminal Kinase) Activation

“…Several studies have suggested that gangliosides are involved in tumor cell adhesion, 26,27) proliferation, 28,29) motility, 29) and metastasis. 30) We also demonstrated that ganglioside GD2 is characteristically expressed on SCLC cells and is involved in proliferation and invasion.…”