Antisense oligodeoxynucleotides downregulated c-sis mRNA expression to inhibit proliferation of vascular smooth muscle cells
Abstract
AIM:
To assess the inhibitory effect of c-sis antisense oligodeoxynucleotides (ODN) on vascular smooth muscle cells (VSMC) proliferation and c-sis proto-oncogene mRNA expression.
METHODS:
The VSMC were cultured with Dulbecco's modified Engle's medium (DMEM) containing synthesized c-sis antisense or sense oligomers, [3H]thymidine (TdR) incorporation into DNA was determined by liquid scintillation counter and the numbers of cells were counted by cell counting plate, the mRNA level was observed by using of reverse transcription-polymerase chain reaction (RT-PCR).
RESULTS:
C-sis antisense ODN 2, 4, 6, 8, and 10 mumol.L-1 inhibited VSMC proliferation (10.3% +/- 0.7%, 22.6% +/- 0.9%, 31.0% +/- 1.1%, 35.4% +/- 0.9%, and 43.3% +/- 1.2%) and [3H]TdR incorporation (6.8% +/- 0.3%, 9.7% +/- 0.7%, 29.0% +/- 0.6%, 32.0% +/- 0.7%, and 50.6% +/- 1.3%) in a concentration-dependent manner. When VSMC were treated with antisense ODN 10 mumol.L-1 for 4 d, the highest inhibitory rate of VSMC growth and that of [3H]TdR incorporation were 60.3% +/- 1.0% and 56.3% +/- 0.9%, respectively. RT-PCR showed that c-sis proto-oncogene antisense ODN downregulated c-sis mRNA expression obviously, while c-sis sense ODN did not inhibit VSMC proliferation and expression of c-sis proto-oncogene mRNA.
CONCLUSION:
The c-sis antisense ODN inhibited VSMC proliferation and downregulated c-sis mRNA level.
Keywords:
To assess the inhibitory effect of c-sis antisense oligodeoxynucleotides (ODN) on vascular smooth muscle cells (VSMC) proliferation and c-sis proto-oncogene mRNA expression.
METHODS:
The VSMC were cultured with Dulbecco's modified Engle's medium (DMEM) containing synthesized c-sis antisense or sense oligomers, [3H]thymidine (TdR) incorporation into DNA was determined by liquid scintillation counter and the numbers of cells were counted by cell counting plate, the mRNA level was observed by using of reverse transcription-polymerase chain reaction (RT-PCR).
RESULTS:
C-sis antisense ODN 2, 4, 6, 8, and 10 mumol.L-1 inhibited VSMC proliferation (10.3% +/- 0.7%, 22.6% +/- 0.9%, 31.0% +/- 1.1%, 35.4% +/- 0.9%, and 43.3% +/- 1.2%) and [3H]TdR incorporation (6.8% +/- 0.3%, 9.7% +/- 0.7%, 29.0% +/- 0.6%, 32.0% +/- 0.7%, and 50.6% +/- 1.3%) in a concentration-dependent manner. When VSMC were treated with antisense ODN 10 mumol.L-1 for 4 d, the highest inhibitory rate of VSMC growth and that of [3H]TdR incorporation were 60.3% +/- 1.0% and 56.3% +/- 0.9%, respectively. RT-PCR showed that c-sis proto-oncogene antisense ODN downregulated c-sis mRNA expression obviously, while c-sis sense ODN did not inhibit VSMC proliferation and expression of c-sis proto-oncogene mRNA.
CONCLUSION:
The c-sis antisense ODN inhibited VSMC proliferation and downregulated c-sis mRNA level.