THE EFFECT OF CYSTEINE PROTEASE INHIBITOR OF SNAKE VENOM(SV-CYSTATIN) ON MOUSE MELANOMA CELLS INVASION IN VITRO AND IN VIVO / 解剖学报

ABSTRACT

Objective To investigate the effect of cysteine protease inhibitor of snake venom(sv-cystatin) on invasiveness and metastasis of melanoma cells.Methods The recombinant plasmid pcDNA3.1/sv-cystatin was constructed and transferred into mouse melanoma cell line B16F1 by lipofection technology.The positive clones were screened by G418 and the sv-cystatin expression was detected by RT-PCR and Western blotting.The ability of tumor cell invasion was identified by Boyden chamber in vitro and tumor invasion animal model in vivo.The ability of tumor cell proliferation and adhension was also determined by MTT assay.Results Expression of sv-cystatin was detected in B16F1/sv-cys cells after gene-transfection.Transfection of sv-cystatin significantly decreased the invasion and migration of B16F1/sv-cys cells along with obviously inhibited the experimental pulmonary metastasis in vivo,but the proliferation and adhension capacity of B16F1/sv-cys cells had no change.Conclusion Transfection of sv-cystatin gene into mouse melanoma cell line results in the suppression of the invasion potential in vitro and in vivo.