[Effects of snake venom cystatin gene on gene expression profiles in mouse melanoma cell line B16F1].

BACKGROUND &

OBJECTIVE

Previously, we have demonstrated that snake venom cystatin (sv-cystatin) plays an important role in tumor invasion and metastasis. This study was to investigate the effects of sv-cystatin on the gene expression profile of mouse melanoma B16F1 cells.

METHODS

pcDNA3.1/sv-cystatin plasmid was constructed and transfected into B16F1 using lipofectamine. Differentially expressed genes between B16F1 transfected with pcDNA3.1/sv-cystatin and pcDNA3.1 were analysed by high throughput microarray technique. Five up-regulated genes and five down-regulated genes were confirmed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS

Out of 1218 transcript species, 45 showed altered expressions: 21 were up-regulated and 24 were down-regulated. These altered genes are involved in cell adhesion and migration, cell immunomodulation, proliferation, differentiation, apoptosis, as well as gene transcription and intra-cellular signal transduction. RT-PCR results of 10 alerted genes were in accordance with the microarray data.

CONCLUSIONS

Sv-cystatin not only inhibits the extracellular matrix, but may also possess other diverse biological functions, including cell immunomodulation, proliferation and differentiation, apoptosis, gene transcription and intra-cellular signal transduction.