Abstract:

Objective To investigate the expression and significance of matrix metalloproteinase -2 (MMP-2) and matrix metalloproteinase -9 (MMP-9) in vitro model of human skin malignant melanoma with MV3 cells. Methods The MV3 malignant melanoma cells were seeded onto de-epidermized dermis (DED) and maintained at the air-liquid interface after submerged culture in two weeks. The reconstructed skin malignant melanoma tissue was examined separately with hematoxylin and eosin(HE) staining and MMP-2、 9 immunohistochemical staining. Furthermore, the expression of MMP-2、MMP-9 mRNA of cultured melanoma organizations was detected by RT-PCR. Results HE staining showed that the growing MV3 cells were in adhesion and attachment to the surface of the DED，aggregated at the concave parts, circularly gathered at cavity of the cutaneous appendages along the BM. It was also observed some of melanoma cells breaking through the BM into DED. Immunohistochemical staining showed that MMP-2、 9 were positive expression along basement membrane of DED in which MV3 melanoma cells were invasive growing. RT-PCR confirmed that the reconstructed malignant melanoma tissue express MMP-2、MMP-9 mRNA, but non MMP-2, 9 mRNA expression in not MV3 melanoma cells inoculated DED organizations. Conclusion The MV3 cells invasion with the expression of MMP-2、 9 in the model of human reconstructed skin malignant melanoma provide an experimental evidence for the study of invasion and metastasis of melanoma in vitro.

Key words: melanoma, in vitro model, matrix metalloproteinases, basement membrane