摘要
目的利用基因芯片研究慢性阻塞性肺疾病(COPD)患者T淋巴细胞氧运输相关基因。方法采集COPD患者和健康人的外周血作为实验血样和对照血样。用Ficoll技术分离外周血淋巴细胞,用分选式流式细胞仪分选并纯化T淋巴细胞,并采用一步法提取总RNA,逆转录合成双链cDNA后,体外转录合成生物素标记的cRNA,片断化后,采用人类全基因表达谱芯片进行芯片杂交,扫描后筛选出差异表达基因。结果初步筛选出COPD/健康人外周血T淋巴细胞氧运输相关高表达差异基因3条。结论基因芯片技术能有效地研究基因的表达谱,筛查出COPD患者T淋巴细胞氧运输相关差异表达基因。
Objective To study the oxygen transporter associated gene expression in thymus-dependent lymopholyte of the COPD patients by gene chips. Methods Peripheral blood of COPD patients（as experimental samples） and of normal man （as control samples） were collected. By Fieoll method, peripheral blood lymphocyte was collected, then thymus-dependent lymphocyte were extracted and puri-fied by flow cytometry. Total RNA were extracted by one-step technique and purified. Then, synthesis double strand eDNA template from total RNA, transcription of cRNA probe with biotin labeling, subsequently, cRNA sample were fragmented. The gene chip was hybridized and scanned. Then fluorescent signal value of gene expressing was obtained, and differential expression genes were sifted. Results 3 oxy-gen transporter associated genes were sifted. Conclusion Gene chip can be applied to study gene expression profiles effectively and to sifte the oxygen transporter genes of the patients with COPD.
出处
《临床肺科杂志》
2009年第6期762-763,共2页
Journal of Clinical Pulmonary Medicine
基金
国家中医药管理局科学技术研究专项课题(02-03JL04)
关键词
慢性阻塞性肺疾病
T淋巴细胞
基因表达
基因芯片
chronic obstructive pulmonary disease
thymus-dependent lymphocyte
gene expression
gene chip
