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Title: Antisense S-oligodeoxynucleotides down-regulate TGF?-production by kupffer cells from CCl4-injured rat livers
Author: Armendariz-Borunda, J.
LeGros Jr., L.
Campollo, O.
Panduro-Cerda, Arturo
Rincon, A.R.
Issue Date: 1997
Abstract: TGF? is a pleiotropic cytokine involved in multiple physiological and pathophysiological regulatory mechanisms. Since TGF? is a disparate modulator of cell recruitment, proliferation and extracellular matrix phenotype for mesenchymal and nonmesenchymal cells, we have been investigating the role of this cytokine in the pathophysiology of liver. In the present paper we investigate which hepatic cell types from CCl4-injured rat livers express TGF? mRNA and produce TGF? in culture, with the aim of further obliterating its biological activity by means of antisense technology. We performed a series of comprehensive molecular studies of in situ hybridization, northern blots, and RT-PCR and we found that only non- parenchymal cells produce TGF? while its expression in hepatocytes was absent. Consistent with the in situ hybridization findings, we observed that Kupffer cells expressed high steady-state levels of TGF? mRNA, while circulating monocytes expressed a smaller amount of TGF? transcripts. We did not detect TGF? gene expression in endothelial cells. These findings were further confirmed by RT-PCR analyses. TGF? activity, as measured by inhibition of [3H]thymidine incorporation by Mv 1 Lu mink lung epithelial cells, was down-regulated in culture by antisense phosphorothioate oligonucleotides. These effects of antisense oligomers were dose-dependent and the sense oligonucleotides had no effect at the same concentration.
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