Suppression of lung metastasis of mouse Lewis lung cancer P29 with transfection of the ganglioside GM₂/GD₂ synthase gene

Abstract

Ganglioside functions in tumor metastasis were analyzed by carbohydrate remodeling of a mouse Lewis lung cancer (subline P29) by introducing ␤1,4GalNAc-T cDNA. Although P29 was originally a low-metastatic subline in the s.c. injection system, it showed high potential in lung metastasis when i.v.-injected via the tail vein. Two lines of GM 2 ؉ transfectants showed markedly reduced metastatic potential to the lung compared to 2 control lines. However, cell proliferation rates and expression levels of various cell adhesion molecules, e.g., integrin family members, SLe x and CD44, were essentially unchanged after transfection of the cDNA. Gangliosides have been considered to play important roles in cell-cell and cell-extracellular matrix recognition and in the modulation of cellular signals on the cell-surface membrane. 1 In particular, gangliosides have been thought to be involved in cell attachment in the case of melanomas, 2-4 neuroblastomas, 3,5,6 gliomas, 7 osteosarcomas 8 and lymphosarcomas. 9 Tumor metastasis results from the successful completion of a series of processes, i.e., detachment and movement of tumor cells from primary sites, invasion into the blood vessel, migration in the bloodstream, adhesion to a vessel wall of a distant organ, invasion across the vessel wall to a secondary site and proliferation to form micro-and macrometastases. 10 One of the most likely situations in which gangliosides are involved in cancer metastasis appears to be adhesion to blood vessels and invasion across vessel walls.Among cancer-associated gangliosides, GM 2 (ganglioside nomenclature is based on that of Svennerholm 10a ) is widely expressed in various epithelial cancer cell types, 11-13 while it is hard to detect GM 2 in normal tissues. In melanoma cells and tissues, GD 3 is commonly and strongly expressed at all stages of the disease 14 -16 and GM 2 /GD 2 appears mainly when the tumors proceed and become more metastatic. 17,18 This is based on novel expression of the GM 2 /GD 2 synthase gene at the advanced phase of the tumor. To clarify the genetic mechanisms for these phenotypic changes of tumors such as melanomas, we isolated the cDNA of ganglioside synthase genes 19 -21 and studied the expression of those glycosyltransferase genes in various tumors. 13,22,23 From this background, we expected that transfection of the GM 2 /GD 2 synthase gene might induce increased metastasis of tumors.To investigate the roles of gangliosides in cancer metastasis in vivo, we used a mouse lung cancer line, Lewis, which has been employed previously in in vivo cancer metastasis studies. 24 P29 is a low-metastatic subline of Lewis lung cancer in the experimental system of s.c. inoculation. 24,25 However, it showed high metastatic potential to the lung when cells were inoculated into the tail vein, suggesting its high adherent activity to endothelia of lung blood vessels.We established GM 2 /GD 2 synthase gene transfectants of P29 and analyzed their metastatic properties; these cells express high levels of GM 2 . Surprisingly, GM 2 -e...

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

“…Also, Hakomori (37) proposed interactions between ganglioside and integrin as one of patterns for signal regulation in the membrane microdomain on the cell surface. We have also suggested the possibility that ganglioside GM2 modulates integrin function (47). Simultaneously, GD3 might also interact with unidentified growth factor receptor(s) and enhance the receptor function, as we demonstrated in the study of a rat pheochromocytoma cell line, PC12 (48).…”

Ganglioside GD3 Enhances Adhesion Signals and Augments Malignant Properties of Melanoma Cells by Recruiting Integrins to Glycolipid-enriched Microdomains

“…Several studies on the modulation of integrins by gangliosides have been reported. For example, membrane cholesterol and sphingolipids have been shown to modify the function of adhesion molecules present on the membrane (25,26). Gangliosides extracted from neuroblastoma cells or atherosclerotic plaques enhance platelet adhesion via integrin binding to collagen (27)(28)(29).…”

“…These microdomains are referred to as lipid rafts or GEM/rafts, and have been considered to be involved in signal transduction, because a variety of signaling molecules, such as Src-family kinases and trimeric G proteins, are also physically associated with the microdomains (48). Moreover, a number of studies have been reported concerning the association of integrins and GEM/rafts (26). For example, the glycosylphosphatidylinositolanchored protein CD24 as a marker for poor prognosis of carcinomas (ovary, breast, and pancreas), induced localization of integrin ␤1 to GEM/rafts with its transfection into A125 and MDA-MB-435S cell lines (49).…”

Nuclear Factor-κB Affects Tumor Progression in a Mouse Model of Malignant Pleural Effusion

“…These cells are characterized by short doubling times in vitro and in vivo and aggressive biological behavior. They can be propagated in wild-type C57B/6 mice, giving rise to lung adenocarcinomas (16)(17)(18). In these studies, we have developed and characterized a new murine model of MPE after instillation of lung adenocarcinoma cells in the pleural space of immunocompetent mice.…”