从石蜡包埋肺癌组织提取RNA检测PML基因转录的研究

PML expression in lung carcinomas: analysis by RT-PCR in formalin-fixed and paraffin-embedded tissue

背景与目的甲醛固定石蜡包埋组织(FPE)包含大量临床病理信息,同时还含有大量的基因和蛋白物质可用来研究临床意义。本研究的目的是探讨从FPE中提取RNA的可能性,检测急性早幼粒细胞白血病基因(PML)在肺癌FPE中的转录表达。方法选取前期经免疫组织化学染色证实无PML蛋白表达的40例肺癌FPE,存档时间在60-85个月,5例新鲜冰冻肺腺癌组织作为对照;利用Trizol一步法提取RNA,经OD值检测验证RNA质量;RT-PCR检测PML的基因转录表达。结果自肺癌FPE和新鲜冰冻组织提取的RNA的OD比值在1.7-2.1。RT-PCR显示,40例FPE以及5例新鲜冰冻组织中,PML(295bp)以及β-actin(318bp)mRNA均有表达。结论利用Trizol一步法自FPE提取RNA进行RT-PCR,可有效扩增出300bp左右的产物;肺癌组织中PML蛋白存在转录后缺失。

Background and objective Formalin-fixed and paraffin-embedded tissue (FPE) with long term follow-up and clinicopathological data are extensively available and easily accessible. They represent an extensive source of genetic materials and proteins to be investigated for clinical usage. We conducted the study to explore the possibility of extracting RNA from FPE of lung cancer, and checking up the transcription levels of the promyelocytic leukemia gene (PML) by RT-PCR. Methods five fresh frozen lung adenocarc...