Specific inhibition of transforming growth factor-beta2 expression in human osteoblast cells by antisense phosphorothioate oligonucleotides.
To elucidate the role of endogenous transforming growth factor (TGF)-beta2 on human osteoblast cell, antisense phosphorothioate oligonucleotides (S-ODNs) complementary to regions in mRNA of TGF-beta2 were synthesized and examined their effects on TGF-beta2 production and cell proliferation in a human osteoblast cell line ROS 17/2. Antisense S-ODNs were designated for three different target regions in the mRNA of TGF-beta2. Among several antisense S-ODN analyzed, an oligonucleotide (AS-11) complementary to the translation initiation site of mRNA of TGF-beta2 demonstrated a selective and strong inhibitory effect on TGF-beta2 production in osteoblast cells. Other antisense S-ODNs which were designated for other regions in mRNA of TGF-beta2 and one- or three-base mismatched analogs of AS-11 showed little or much less antisense activities than AS-11. Therefore, the most effective target site in mRNA of TGF-beta2 is at the initiation codon region. The antisense effects of AS-11 were observed without reduction of levels of mRNA of TGF-beta2. Furthermore, the inhibition of TGF-beta2 expression by antisense S-ODN appeared to enhance cell proliferation, demonstrating the growth inhibitory effect of autocrine TGF-beta2 in osteoblast cells.
Shen
Z J
ZJ
Department of Biochemistry, Kyungpook National University, Taegu, Korea.
Kim
S K
SK
Kwon
O S
OS
Lee
Y S
YS
Moon
B J
BJ
D015152:Blotting, Northern; D015153:Blotting, Western; D002411:Cation Exchange Resins / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D003062:Codon; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D050356:Lipid Metabolism; D008055:Lipids; D016376:Oligonucleotides, Antisense / Q000378:metabolism* / Q000493:pharmacokinetics; D010006:Osteoblasts / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000096:biosynthesis*; D053781:Transforming Growth Factor beta2
D002411:Cation Exchange Resins; D003062:Codon; D007202:Indicators and Reagents; D008055:Lipids; C086724:Lipofectamine; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C509041:TGFB2 protein, human; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2