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Objective To study the effect of endogeneous ganglioside (GS) on the adhesion of LA-N5 neuroblastoma cells to collagen. Methods LA-N5 cells were cultured in the presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol (D-PDMP), an inhibitor of glucosylceramide synthase. The adhesion of the LA-N5 cells to immobilized collagen was tested. Results After 6 days, endogenous GS was reduced by 98%. No change in the cell morphology, viability, proliferation rate or percentage of apoptotic cells was observed. The adhesion to collagen of cells exposed to D-PDMP was reduced by 65% compared to the control LA-N5 cells: OD 570 0.07± 0.01 vs 0.21± 0.030(P< 0.01). When GS-depleted cells were pre-incubated in a conditioned medium collected from the control cells, the adhesion to collagen was restored and was comparable to that of the control cells (P> 0.05). Similarly, the pre-incubation of GS-depleted cells with purified tumor GS GD 2, the most abundant GS in LA-N5 cells, restored adhesion. Conclusions Endogenous tumor GS regulates neuroblastoma cell adhesion to collagen, suggesting that it may play a role in tumor cell migration, invasion and metastasis.
Methods of LA-N5 cells were cultured in the presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline- Results of 6 days, endogenous GS was reduced by 98%. No change in the cell morphology, viability 1-propanol (D-PDMP), an inhibitor of glucosylceramide synthase. The adhesion of the LA-N5 cells to immobilized collagen was tested. The proliferation rate or percentage of apoptotic cells was observed. The adhesion to collagen of cells exposed to D-PDMP was reduced by 65% compared to the control LA-N5 cells: OD 570 0.07± 0.01 vs 0.21± 0.030 (P< 0.01) . When GS-depleted cells were pre-incubated in a conditioned medium collected from the control cells, the adhesion to collagen was restored and was comparable to that of the control cells (P> 0.05). Similarly, the pre-incubation of GS- Depleted cells with purified tumor GS GD 2, the most abundant GS in LA-N5 cells, Conclusion chemotherapy. Conclusions Endogenous tumor GS regulates neuroblastoma cell adhesion to collagen, suggesting that it may play a role in tumor cell migration, invasion and metastasis.