Inhibitory effects of antisense oligonucleotides on the expression of procollagen type III gene in mouse hepatic stellate cells transformed by simian virus 40.
The effects of phosphorothioate antisense oligonucleotides (ASO), complementary to the AUG start region, the junctional region of the intron and exon, and to exon of the procollagen type III gene, were investigated in a mouse hepatic stellate cell (HSC) line transformed by the simian virus 40 gene, SV68c-IS cells. ASO were transfected by lipofection. Immunohistochemistry, western and northern blotting showed inhibitory effects on procollagen type III gene expression by ASO that were complementary to the AUG start region and the junctional region of the intron and exon 2. However, ASO complementary to the exon 2 and 3, junctional region of the intron and exon 3, and sense oligonucleotides complementary to each ASO did not show any inhibitory effects. The effects of ASO complementary to the AUG start region were greater than those of ASO complementary to the junctional region. The effects of ASO were transient and a large amount of ASO was required to induce inhibitory effects without lipofection. ASO were effective in inhibiting the expression of the procollagen type III gene in the HSC which is well known to play a critical role in liver fibrosis.
Horie
S
S
Second Department of Pathology, Faculty of Medicine, Tottori University, Yonago, Japan.
Kitamura
Y
Y
Kawasaki
H
H
Terada
T
T
D000199:Actins / Q000032:analysis; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000032:analysis; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D016373:DNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D003893:Desmin / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D007150:Immunohistochemistry; D008099:Liver / Q000737:chemistry / Q000166:cytology* / Q000821:virology; D051379:Mice; D009130:Muscle, Smooth / Q000737:chemistry; D011347:Procollagen / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013539:Simian virus 40 / Q000254:growth & development*; D013997:Time Factors
D000199:Actins; D000957:Antigens, Viral, Tumor; D016373:DNA, Antisense; D003893:Desmin; D005904:Glial Fibrillary Acidic Protein; D011347:Procollagen; D012333:RNA, Messenger