[TGF-beta 1 antisense gene transfer into ito cells and suppressed extracellular matrix production].

OBJECTIVE

To investigate possible role of antisense TGF-beta 1 RNA in the regulation of TGF-beta 1 and ECM production in Ito cells.

METHOD

A human transforming growth factor-beta 1(TGF-beta 1) cDNA (1467 bp) was inserted in reverse orientation into the retroviral vector, constructed retroviral vector pLATSN of antisense RNA for TGF-beta 1. A higher-titer, recombinant retroviral vector carried antisense RNA for TGF-beta 1 produced in PA317 packaging cells has been introduced into human Ito cells lines LI90. After selection with G418, resistant colonies were obtained.

RESULTS

Stable integration of retrovirus in infectants was shown the presence of antisense RNA was detected by RT-PCR. The expression of TGF-beta 1 protein and the production of extracellular matrix such as FN, Co1A1 were markedly decreased in the antisense TGF-beta 1 transfected cultured cells by ELISA, immunohistochemistry and in situ hybridization.

CONCLUSION

Antisense RNA of TGF-beta 1 can be successfully used to inhibit Ito cells activated, endogenous TGF-beta 1 mRNA and extracellular matrix produced, and may provide a basis for the development of anti-fibrosis gene therapy.