Inhibitory effect of tissue transglutaminase (tTG) antisense oligodeoxynucleotides on tTG expression in cultured bovine trabecular meshwork cells

Abstract

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-ASDON₁ 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.

Keywords

tissue transglutaminase antisense oligodeoxynucleotide / trabecular meshwork cell / culture

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