Comparison of Fixation Methods for Preservation of Morphology, RNAs, and Proteins From Paraffin-Embedded Human Cancer Cell-Implanted Mouse Models
Comparison of Fixation Methods for Preservation of Morphology, RNAs, and Proteins From Paraffin-Embedded Human Cancer Cell-Implanted Mouse Models
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DOI：
10.1369/jhc.2010.957217
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发表时间：
2011-01-01
影响因子：
3.2
通讯作者：
Naito, Zenya
中科院分区：
文献类型：
--
作者：
Matsuda, Yoko;Fujii, Takenori;Naito, Zenya
将人肿瘤细胞异种移植到免疫缺陷小鼠体内是阐明特定分子或化学物质在体内作用的重要方法。最近，这种方法已被报道为确定性检查，以确定肿瘤干细胞。在这项研究中，作者比较了植入人宫颈癌细胞的裸鼠石蜡包埋组织的形态以及核糖核酸（RNA）和蛋白质的质量和数量，然后用常用的固定剂固定，包括4%多聚甲醛（PFA），10%中性缓冲福尔马林（NBF），20% NBF和99%乙醇（EtOH）。从PFA和NBF固定的石蜡包埋组织中分离的RNA质量较高，而EtOH固定的组织显示RNA降解。NBF固定的组织表现出良好的形态质量，但乙醇固定的组织显示细胞收缩。免疫组化结果显示差异取决于固定。99%EtOH固定的样品显示Ki-67和VEGF-A免疫反应性降低，但细胞角蛋白免疫反应性提高。本研究表明，对于石蜡包埋癌细胞植入模型中的RNA保存，福尔马林固定比酒精固定更好。免疫组化结果根据固定材料和抗体而显著不同;因此，需要合适的固定来量化和比较癌细胞植入裸鼠组织的免疫组化染色结果。（J Histochem Cytochem 59：68-75，2011）
Xenograft transplantation of human tumor cells into immunodeficient mice is an important method to clarify the roles of specific molecules or chemicals in vivo. Recently, this method has been reported as a definitive examination to identify tumor stem cells. In this study, the authors compared the morphology and the quality and quantity of ribonucleic acid (RNA) and protein in paraffin-embedded tissues of nude mice implanted with human uterine cervical cancer cells, followed by fixation with commonly used fixatives, including 4% paraformaldehyde (PFA), 10% neutral buffered formalin (NBF), 20% NBF, and 99% ethanol (EtOH). The quality of the isolated RNA from PFA- and NBF-fixed paraffin-embedded tissues was high, while EtOH-fixed tissues showed degradation of RNA. NBF-fixed tissues showed excellent quality of morphology, but EtOH-fixed tissues showed contraction of cells. Immunohistochemical results showed differences depending on fixations. The 99% EtOH-fixed samples showed decreases of Ki-67 and VEGF-A immunoreactivities, but improved cytokeratin immunoreactivity. This study indicated that formalin fixation is better than alcohol fixation for RNA preservation in paraffin-embedded cancer cell implantation models. Immunohistochemical results differed markedly depending on fixation materials and antibodies; therefore, suitable fixations are needed to quantify and compare the results of immunohistochemical staining on cancer cell implanted nude mice tissues. (J Histochem Cytochem 59:68-75, 2011)