|Inhibitory effect of retroviral vector containing anti-sense Smad(4) gene on Ito cell line, LI90|
|Xu, XB; Leng, XS; He, ZP; Liang, ZQ; Lin, K; Wei, YH; Yu, X; Peng, JR|
|关键词||genetic vectors retroviridae gene therapy DNA antisense hepatic stellate cell|
|刊名||CHINESE MEDICAL JOURNAL|
|WOS标题词||Science & Technology|
|类目[WOS]||Medicine, General & Internal|
|研究领域[WOS]||General & Internal Medicine|
|关键词[WOS]||HEPATIC STELLATE CELLS ; GROWTH-FACTOR-BETA ; LIVER FIBROSIS ; TGF-BETA ; IN-VITRO ; RAT ; EXPRESSION ; ACTIVATION ; RECEPTOR ; PROLIFERATION|
Background Transforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad(4) is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad(4) gene on Ito cell line, L190.
Methods The recombinant retroviral vector pLXSN-Smad(4) was constructed by cloning the rat antisense Smad(4) cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad(4) and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) L190 were infected with these retroviruses followed by selection with G418. The expression of Smad(4) was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, H-3-TdR and H-3-proline uptake by HSCs and the production of extracellular matrix were assessed.
Results mRNA and protein expressions of Smad(4) in L190 cells transfected with retrovirus containing the antisense Smad(4) gene were much lower than those in L190 cells transfected with empty vector or parental L190 cells. Cells hypoexpressing the Smad(4) gene exhibited a slower rate of growth, a lower uptake of H-3-TdR and H-3-proline (P < 0.01), and smaller production of th extracellular matrix, compared with parental L190 cells and cells transfected with empty retrovirus.
Conclusions The antisense Smad, gene can suppress the expression of the Smad(4) gene, reduce endogenous production of Smad(4) mRNA and protein, block TGF-beta1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM). Our results may provide a basis for the development of antifibrotic gene therapy.
|作者单位||1.Beijing Univ, Hepatobiliary Surg Ctr, Peoples Hosp, Beijing 100044, Peoples R China|
2.AF Gen Hosp, Beijing 100036, Peoples R China
3.Third Mil Med Univ, Hepatobiliary Surg Ctr, SW Hosp, Chongqing 400038, Peoples R China
|Xu, XB,Leng, XS,He, ZP,et al. Inhibitory effect of retroviral vector containing anti-sense Smad(4) gene on Ito cell line, LI90[J]. CHINESE MEDICAL JOURNAL,2004,117(8):1170-1177.|
|APA||Xu, XB.,Leng, XS.,He, ZP.,Liang, ZQ.,Lin, K.,...&Peng, JR.(2004).Inhibitory effect of retroviral vector containing anti-sense Smad(4) gene on Ito cell line, LI90.CHINESE MEDICAL JOURNAL,117(8),1170-1177.|
|MLA||Xu, XB,et al."Inhibitory effect of retroviral vector containing anti-sense Smad(4) gene on Ito cell line, LI90".CHINESE MEDICAL JOURNAL 117.8(2004):1170-1177.|