Abstract:

Objective

To study the inhibition of the invasiveness of ameloblastoma cells by negative regulation of MMP-2 gene using RNA interference.

Methods

Ameloblastoma cells were cultured in Defined Keratinocyte-SFM medium in vitro and MMP-2 was detected by immunofluorescence.Plasmid-vector for MMP-2 targeted siRNA was constructed and transfected into ameloblastoma cells. Laser confocal microscope was used to detect the positive transfected cells. Flowocytometer was used to analyze the transfection rate.Reverse transcription polymerase chain reaction（RT-PCR） analysis was performed to detect the expression of MMP-2 mRNA. Western-blot analysis was performed to investigate MMP-2 protein. Transwell analysis was done to explore the invasion of ameloblastoma cells. The analysis of variance （ANOVA） was used to analyze the data.

Results

Positive expression of MMP-2 was observed in ameloblastoma. The transfection rate of plasmid-vector for siRNA into ameloblastoma cells was 63.6%. After transfection with MMP-2 targeted siRNA， the expression of MMP-2 mRNA and MMP-2 protein decreased 69.3% and 64.2%， respectively， P＜0.05. The inhibition rate against the invasion of ameloblastoma cells was 61.2%.

Conclusion

MMP-2 targeted plasmid-vector for siRNA is successfully transfected into ameloblastoma cells and MMP-2 gene is silenced by siRNA. MMP-2 has close relation with the invasiveness of ameloblastoma cells.

Key words: Matrix metalloproteinases-2, RNA interference, Small interference RNA（siRNA）, Gene silence, Inhibition