贵州医科大学学报

2005, (03) 241-243

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石蜡包埋组织中RNA提取方法的改进和完善
The Improvement of Isolation Technique of RNA from Formalin-fixed and Paraffin-embedded Tissues

孟青,杨文秀,胡军,高勤
MENG Qing, YANG Wen-xiu, HU Jun, GAO Qin (Department of Pathology, Guiyang Medical College, Guiyang 550004, China)

摘要(Abstract):

目的:建立和完善石蜡组织RNA提取的方法。方法:收集临床病理诊断用石蜡包埋组织块共100例,经RNA灭活处理后,进行组织切片、脱蜡、清洗、消化等一系列提取前预处理,用TRIZOL试剂提取组织的RNA,并在-20℃长时间沉淀RNA。对提取的RNA以核酸测定仪检测RNA的浓度和纯度,并通过RT-PCR检查提取RNA的β-actin基因,以检测提取RNA的质量。结果:100例石蜡包埋组织中提取的RNA经过核酸测定仪测定后,浓度在20~50mmol/L,260nm及280nm光密度比在1.75~2.0;经过RT-PCR反应,提取的RNA都能扩增出190bp的β-actin目的片段。结论:通过RNA提取前对材料的一系列合理处理,从福尔马林固定、石蜡包埋档案组织材料中提取小片段RNA进行目的基因mRNA检测,可以成为一种稳定成熟的方法应用于临床病理诊断。
Objective: To improve the technique for extracting RNA from formalin-fixed and paraffin-embedded tissue. Methods: Samples of 100 cases that diagnosed pathologically were collected. After some pretreatment such as deparaffinage, washing, digestion, total RNA were extracted from these tissues with TRIZOL reagent and, then, kept at -20 ℃for long time (above 2 h) to settle RNA. The pureness and content of these RNA extracts were assayed with a nucleic acid detecting instrument and the quality of the extracts was tested with detecting of gene β-actin with RT-PCR. Results: The concentration of RNA in the extracts were between 20~50 mmol/L.The ratio of 260 nm/280 nm were between 1.75~2.00. The RNA of β-actin were detected in all cases. Conclusions: The reasonable pretreatment adopted in this study makes it feasible to extract RNA from formalin-fixed and paraffin-embedded tissue and to detect genes specific to some diseases in daily pathological diagnosis.

关键词(KeyWords): RNA,肿瘤;石腊包埋;逆转录聚合酶链反应
RNA, neoplasm; paraffin embedding; reverse transcriptase polymerase chain reaction

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作者(Author): 孟青,杨文秀,胡军,高勤
MENG Qing, YANG Wen-xiu, HU Jun, GAO Qin (Department of Pathology, Guiyang Medical College, Guiyang 550004, China)

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