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[TGF-beta 1 antisense gene transfer into ito cells and suppressed extracellular matrix production]
1Hepatobiliary Surgery Center, Southwestern Hospital, Third Military Medical University, Chongqing.
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.