摘要
目的比较微小病变型肾病综合征(MCNS)患儿与正常健康儿童外周血单个核细胞(PBMC)的基因表达谱变化,筛查MCNS相关致病基因,为揭示MCNS的发病分子机制和临床治疗提供线索。方法原发性MCNS患儿7例,正常同年龄对照组7例,Trizol法抽提PBMC总RNA;采用人类基因组表达谱芯片检测MCNS及正常健康儿童PBMC基因mRNA水平;采用逆转录-聚合酶链反应(RT-PCR)和荧光定量PCR检测部分基因转录水平,鉴定芯片相关检测结果。结果在33 000个基因转录本中,有969个转录本在MCNS患儿PBMC中存在表达差异,其中表达上调552个,表达下调417个。RT-PCR和荧光定量PCR检测结果与基因芯片较为一致。结论采用人类基因组表达谱芯片可快速有效地检测MCNS患儿PBMC中基因表达谱的变化,进而证实MCNS的发生、发展是涉及多基因改变的复杂过程。
Objective To screen for the causative genes involved in the occurrence and development of minimal changes nephritic syndrome(MCNS) and to furtherly assist the genetic diagnosis and treatment of MCNS. Methods Human genome U133 Array Set from Affymetrix lnc was used to evaluate gene expression patterns in peripheral blood mononuclear celIs(PBMC) isolated from 7 children with primary MCNS and 7 age- matched health volunteers. Reverse transcription- polymerase chain reaction(RT- PCR) and real - time PCR were performed to identify the findings of gene chip. Results Of 33 000 genes detected, 969 genes showed significant difference between children with MCNS and healthy volunteers; 552 genes were up - regulated,while 417 genes down - regulated significantly. Findings from RT- PCR and real - time PCR were consistent with those of gene chip. Conclusions Gene chip of expression patterns is a powerful method to detect expression difference of genes correlated with MCNS. Occurrence and development of MCNS can be a complicated proceas that many correlative genes may participate in.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2006年第19期1317-1319,共3页
Journal of Applied Clinical Pediatrics
基金
广东省医学科研基金项目资助(A2002657)
关键词
肾病综合征
微小病变型
基因表达谱
儿童
minimal change nephritic syndrome
gene expression pattern
child