微阵列预测分析法筛选乙型肝炎肝硬化发生的风险基因

肖春阳; 李沁恺; 于景霞; 刘婷; 陆伦根; 徐铭益

doi:10.3969/j.issn.1001-5256.2016.07.022

微阵列预测分析法筛选乙型肝炎肝硬化发生的风险基因

DOI: 10.3969/j.issn.1001-5256.2016.07.022

上海交通大学附属第一人民医院消化科

基金项目:

国家科技部“十二五”重大专项(2013ZX10002004-002-003)；国家自然基金面上项目(81570547)；院优秀青年培养计划(061405)；

详细信息

中图分类号: R512.62;R575.2
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出版历程
- 出版日期: 2016-07-20

Risk genes for the development of chronic hepatitis B cirrhosis assessed by prediction analysis of microarrays

Research funding:

摘要

摘要: 目的采用基因芯片技术筛选预测乙型肝炎肝硬化发生的风险基因。方法收集2008年4月-2010年12月于上海交通大学附属第一人民医院就诊的慢性乙型肝炎（CHB）患者40例,建立蕊片筛选风险基因队列（分为5组:S0、S1、S2、S3、S4;每组8例）,肝活组织病理学检查确定肝纤维化分期（以Scheuer病理评分为标准）,另留取临床资料和肝组织样本。采用人Affymetrix基因芯片技术检测CHB患者肝组织的基因表达谱,微阵列显著分析（SAM）和微阵列预测分析（PAM）筛选预测肝硬化发生的风险基因组。实时定量PCR验证风险基因mRNA在肝组织中的表达情况。分类资料采用χ2检验进行比较;符合正态分布的连续变量比较采用t检验和单因素方差分析,进一步两两比较采用SNK-q检验;不满足正态分布的采用Mann-Whitney U秩和检验。结果Affymetrix基因芯片共筛选出1674个差异表达基因,差异基因聚类分析显示肝纤维化分期不同,组间的基因表达也存在差异,从而提示基因表达谱与组织纤维化分期存在较好的一致性。以4种不同的分类法分析,SAM筛选出87个显著基因,进而采用PAM筛选出14个"高风险"基...
- 肝炎,乙型,慢性 /
- 肝硬化 /
- 基因表达 /
- 微阵列分析
Abstract: Objective To investigate the risk genes for predicting the development of chronic hepatitis B( CHB) cirrhosis using gene chip technology. Methods A total of 40 CHB patients who visited Shanghai First People's Hospital from April 2008 to December 2010 were enrolled as a clinical cohort and were divided into S0,S1,S2,S3,and S4 groups,with 8 patients in each group. Liver biopsy was performed to determine fibrosis stage with the Scheuer pathological score as the criteria,and clinical data and liver tissue samples were reserved. The Human Affymetrix Gene Chip was used to establish the gene expression profiles of liver tissues in CHB patients,and the significance analysis of microarrays( SAM) and prediction analysis of microarrays( PAM) were used to screen out the risk genomes for predicting the development of CHB cirrhosis. Quantitative real- time PCR was used to measure the mRNA expression of risk genes in liver tissue. The chi- square test was used for comparison of categorical data. The t- test and a one- way analysis of variance were used for comparison of normally distributed continuous data,and SNK- q test was used for further comparison between any two groups; the Mann- Whitney U rank sum test was used for comparison of non- normally distributed continuous data. Results A total of 1674 differentially expressed genes were screened out by Affymetrix Gene Chip. A cluster analysis of these genes showed that gene expression showed differences between groups with different fibrosis stages,which suggested that the gene expression profile was well consistent with fibrosis stage. Four different classification methods were used for analysis,and 87 significant genes were screened out by SAM and 14 “high- risk”genes were screened out by PAM. The quantitative real- time PCR showed the expression of 6 risk genes( CD24,CXCL6,EHF,ITGBL1,LUM,and SOX9) differed significantly between groups S0,S1- 3,and S4( P < 0. 05),and the S1- 3 and S4 groups showed significantly upregulated expression of these genes compared with the S0 group( all P < 0. 05). Conclusion The 6 high- risk factors screened out and verified by gene chip technology help to predict the probability of developing liver cirrhosis in CHB patients and can be used as the diagnostic genes for predicting hepatitis B cirrhosis.
- hepatitis B /
- chronic /
- liver cirrhosis /
- gene expression

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