Inhibitory effect of tissue transglutaminase (tTG) antisense oligodeoxynucleotides on tTG expression in cultured bovine trabecular meshwork cells.
To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides (tTG-ASDON) on tTG expression in cultured bovine trabecular meshwork cells (BTMCs) in vitro and explore a new treatment alternative for primary open angle glaucoma (POAG), the ASDON1 and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique-Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON and tTG-ASDON2 were significantly decreased as compared with that of the controls (P < 0.05). On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down-regulated by tTG- ASDON. As a result, tTG-ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG.
Hu
Yizhen
Y
Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Zhang
Haijiang
H
Xiong
Xinchun
X
Cao
Yang
Y
Han
Yongjuan
Y
Xi
Zulian
Z
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D019204:GTP-Binding Proteins; D005902:Glaucoma, Open-Angle / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014129:Trabecular Meshwork / Q000166:cytology / Q000378:metabolism*; D011503:Transglutaminases / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins