| ObjectiveThe plasma membrane glycoprotein CD147(also known as Basigin or EMMPRIN) is a multifunctional member of the immunoglobulin superfamily. The previous studies have shown that CD147was involved in melanoma cell proliferation, invasiveness, metastasis and angiogenesis, while the mechanism has not been very clear. The results of many studies have demonstrated that the expression of glucose transporters,especially GLUT1(glucose transporter member1), is increased in a variety of malignancies. This study was undertaken to determine the association between CD147and GLUT1, then to investigate the interaction region, location and the function of glycolysis in melanoma cells, in order to confirm the molecular mechanisms of malignant melanoma.Methods1. Define the correlation between CD147and GLUT1in human malignant melanoma tissue microarray.2. The location of CD147and GLUT1in A375cells was detected by cell immunofluorence.3. The eukaryotic expression vectors of GLUT1, full length and mutants of CD147were constructed.4. The co-immunoprecipitation was used to verify the interaction between CD147and GLUT1, and confirm the interaction site.5. Form tumor with A375cells and A375-shCD147cells in nude mice.18F-FDG Micro-PET/CT scan was performed to evaluate the ability of glucose uptake.Results1. There showed a positive correlation between GLUT1and CD147in human malignant melanoma tissue microarray.2. The co-immunoprecipitation showed that CD147was co-localized with GLUT1on cell membrane.3. The eukaryotic expression vectors of GLUT1, full length and mutants of CD147were successfully constructed.4. The interaction between GLUT1and CD147was detected by co-IP, and the second IgG like domain as the interaction site.5. The CD147knocked-down tumor showed a weaker ability of glucose uptake.ConclusionsThere showed a positive correlation between GLUT1and CD147. CD147interact with GLUT1in its second IgG like domain.CD147had certain up-regulation on Glucose uptake of A375cells. |
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