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首页> 外文期刊>Biopreservation and biobanking >Surgical Specimens of Colorectal Cancer Fixed with PAXgene Tissue System Preserve High-Quality RNA
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Surgical Specimens of Colorectal Cancer Fixed with PAXgene Tissue System Preserve High-Quality RNA

机译：用PAXgene组织系统固定的结直肠癌手术标本保留了高质量的RNA

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Background: RNA analysis of surgical specimens is one of the most useful methods for exploring biomarkers of advanced cancer. The most readily available source for RNA is formalin-fixed, paraffin-embedded (FFPE) specimens, but RNA isolated from FFPE tissue is of limited use. The PAXgene Tissue (PAX) system is a formalin-free system designed to improve the quality of molecular analysis without diminishing the quality of histopathological analysis. In this human colorectal cancer tissue study, we aimed to evaluate whether surgical specimens fixed with PAX can preserve high-quality RNA in comparison with FFPE and fresh-frozen tissue specimens. Methods: Ten consecutive advanced colorectal cancer patients undergoing colectomy were examined. Each specimen was processed in three ways: as frozen tissue, as PAX-fixed tissue, and as formalin-fixed tissue. RNA integrity numbers (RINs) were assessed using an Agilent Bioanalyzer. RNA transcript levels and stability were investigated by quantitative real-time PCR. We also evaluated the immunohistochemical intensity of Ki-67, CEA, and EGFR in the PAX samples. Results: The average RINs of RNA extracted from frozen and PAX samples were significantly higher than those from FFPE samples (p<0.001). The cycle threshold (Ct) values were similar in PAX and frozen samples, but significantly increased in FFPE samples (p<0.001). Most of the Ct values in the PAX samples did not differ significantly from those in the matched frozen samples. On the other hand, most of the Ct values in the FFPE samples differed significantly from those in the matched frozen samples. The immunohistochemical intensity in the PAX samples was well preserved. Conclusions: The quality of RNA extracted from PAX samples may be slightly inferior to that from frozen samples, but is greatly superior to that from FFPE samples.
机译：背景：手术标本的RNA分析是探索晚期癌症生物标志物最有用的方法之一。最容易获得的RNA来源是福尔马林固定，石蜡包埋（FFPE）的标本，但从FFPE组织中分离的RNA用途有限。 PAXgene Tissue（PAX）系统是一种不含福尔马林的系统，旨在提高分子分析的质量而又不降低组织病理学分析的质量。在这项人类大肠癌组织研究中，我们旨在评估与FFPE和新鲜冷冻的组织标本相比，用PAX固定的手术标本是否可以保留高质量的RNA。方法：对十名连续接受结肠切除术的晚期大肠癌患者进行检查。每个标本以三种方式处理：作为冷冻组织，作为PAX固定的组织和作为福尔马林固定的组织。使用安捷伦生物分析仪评估RNA完整性数（RINs）。通过定量实时PCR研究RNA转录水平和稳定性。我们还评估了PAX样品中Ki-67，CEA和EGFR的免疫组织化学强度。结果：从冷冻和PAX样品中提取的RNA的平均RINs显着高于从FFPE样品中提取的RNA（p <0.001）。在PAX和冷冻样品中，循环阈值（Ct）值相似，但在FFPE样品中，循环阈值（Ct）显着增加（p <0.001）。 PAX样品中的大多数Ct值与匹配的冷冻样品中的Ct值没有显着差异。另一方面，FFPE样品中的大多数Ct值与匹配的冷冻样品中的Ct值显着不同。 PAX样品中的免疫组织化学强度得到了很好的保存。结论：从PAX样品中提取的RNA的质量可能稍逊于从冷冻样品中提取的RNA，但大大优于从FFPE样品中提取的RNA。

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