Attenuation of α1 Collagen Production with Antisense Ribonucleic Acid in Cultured Hypertrophic Scar Fibroblasts

Abstract

Background: It has been demonstrated that hypertrophic scar fibroblasts (HSFs) overexpress collagen messenger ribonucleic acid (mRNA) and protein, especially α1 collagen. Antisense nucleic acids are effective in inhibiting harmful or uncontrolled gene expression, suggesting that antisense ribonucleic acid (RNA) can effectively downregulate the expression of α1 collagen gene and attenuate the scars. Aims: This study was conducted to observe the effect of recombinant plasmid pREP9-COL1 on α1 collagen expression in HSFs and clarify the prospect of antisense RNA on scar treatment. Methods: The α1 collagen gene fragment including the region of 5′ UTR to exon (229 bp) was cloned in the eukaryotic expression plasmid pREP9 in the antisense orientation relative to the RSV-LTR promoter to reconstruct the pREP9- COL1 plasmid. Then it was transferred into HSFs through lipofectamine. The expression of α1 collagen was examined by immunostaining, reverse-transcriptase polymerase chain reaction, and Western blots. Results: The recombinant plasmid pREP9-COL1 with a correct sequence was constructed successfully; pREP9-COL1 consistently inhibited human α1 collagen gene expression at both mRNA and protein levels. Conclusions: Antisense RNA was effective in downregulating α1 collagen expression of HSFs. Therefore, this approach offered a prospect of scar treatment by attenuation of α1 collagen production with antisense RNA.