Abstract:

Objectives

This study was designed to evaluate the probability of applying TGFβ3 small interfering RNA （siRNA）to transfect cultured palatal process cells.

Methods

Chemically synthetical TGFβ3 siRNA warped by Lipofectamine^TM2000 were used to transfect primary cultured murine embryonic epithelial and mesenchymal cells， which were derived from gestational day 14 Bal b/c mouse embryos and examined by phase-contrast microscope and MTT.Semi-quantitative RT-PCR and western blot were used to exam the gene expression of TGFβ3，Smad2， BMP2 at 24， 48， 72 hours after using standard dosage Stealth^TM RNAi.

Results

After using standard dosage of Stealth^TM RNAi 24， 48， 72 hours， the TGFβ3 gene expression was down regulated significantly at 24， 48， 72 hours after treatment. The Smad2 gene expression was down regulated significantly at 24 hours after treatment. However， the expression increased but not recovered at 48 hours and 2 hours after. But BMP2 gene expression showed no significant change at all three time points.

Conclusions

TGFβ3 Stealth^TM RNAi could silence TGFβ3 gene effectively in the murine embryonic palate cells. However， the silencing effect had tendency to attenuate. Smad2 gene mRNA and the protein expression decreased along with TGFβ3 gene silence， but BMP2 gene expression had non-significance change.

Key words: Cleft palate, TGFβ3, Smad2, Signal pathway, RNA interference