摘要
目的:筛查肝纤维化组织相关基因,探讨肝纤维化发生机制.方法:抽提肝纤维化组织和正常肝组织总RNA来制备探针,经杂交、洗涤后,通过计算机扫描分析肝纤维化组织和正常肝组织基因表达谱的差异情况,并用实时荧光定量PCR技术对其中部分差异基因的表达水平变化进行验证.结果:筛选出差异表达的基因68个,其中肝纤维化组织中表达上调的基因35个,表达下调的基因33个,这些基因按照功能可以分为调控细胞信号转导、DNA损伤与修复、转录调控因子、代谢相关基因以及未知功能基因.与正常肝组织比较,肝纤维化组织中C/EB Pβ mRNA表达下调(22.02±4.82vs59.13±8.21,P<0.01),MMP-14 mRNA表达上调(257.33±26.58vs21.65±4.37,P<0.01),结果与基因芯片一致.结论:多种基因参与肝纤维化的形成过程,肝纤维化组织与正常肝组织之间存在有明显的基因表达差异.
AIM: To screen the genes associated with human hepatic fibrosis and explore the mechanism of hepatic fibrosis.
METHODS: Total mRNA was obtained from human hepatic fibrosis tissues and normal liver tissues respectively to prepare probe. After hybridization and cleaning, computer scan technique was used to analyze the gene expression maps of liver fibrosis tissues and normal liver tissue. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was adopted to verify the expression changes of some differential genes.
RESULTS: Sixty-eight differential genes were screened out, of which 35 were up-regulated and 33 were down-regulated. These genes were categorized according to their functions as cell signal transmission-regulating genes, DNA impairment and repair genes, translation-regulating factors, metabolism-related genes and functionunknown genes. As compared with that in normal liver tissues, C/EBPβ mRNA expression was down-regulated (22.02 ± 4.82 vs 59.13 ± 8.21, P 〈 0.01), while MMP-14 mRNA expression was up-regulated (257.33 ± 26.58 vs 21.65 ± 4.37, P 〈 0.01). This conformed to the results of cDNA microarray.
CONCLUSION: Multiple genes are involved in the formation of hepatic fibrosis and there is a significant difference in gene expression between normal and fibrotic liver tissues.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第14期1525-1529,共5页
World Chinese Journal of Digestology
基金
贵州省优秀青年科技人才基金资助项目
No.20050518
贵州省省长基金资助项目
No.2005223
贵州省卫生厅科技基金资助项目
No.2006046~~
关键词
肝纤维化
肝组织
基因芯片
基因表达
实时荧光定量PCR技术
Hepatic fibrosis
Liver tissue
cDNA microarray
Gene expression
Real-time fluorescence quantitative polymerase chain reaction