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学科主题: 临床医学
题名:
Inhibitory effect of retroviral vector containing anti-sense Smad(4) gene on Ito cell line, LI90
作者: 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
发表日期: 2004-08-01
卷: 117, 期:8, 页:1170-1177
收录类别: SCI
文章类型: Article
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
英文摘要:

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.

语种: 英语
WOS记录号: WOS:000223711500009
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/61296
Appears in Collections:北京大学第二临床医学院_期刊论文

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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

Recommended Citation:
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.
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