Delfour Christophe, Roger Pascal, Bret Caroline, Berthe Marie-Laurence, Rochaix Philippe, Kalfa Nicolas, Raynaud Pierre, Bibeau Frédéric, Maudelonde Thierry, Boulle Nathalie
Laboratoire d'Anatomie Pathologique, Centre Hospitalier Universitaire de Montpellier, Hôpital Arnaud de Villeneuve, France.
J Mol Diagn. 2006 May;8(2):157-69. doi: 10.2353/jmoldx.2006.050105.
Methacarn and RCL2, a new noncrosslinking fixative, were compared to formalin-fixed or frozen tissue samples of the same invasive breast carcinoma and were evaluated for their effects on tissue morphology and immunohistochemistry as well as DNA and RNA integrity. The histomorphology of methacarn- or RCL2-fixed paraffin-embedded tumors was similar to that observed with the matched formalin-fixed tissues. Immunohistochemistry using various antibodies showed comparable results with either fixative, leading to accurate breast tumor diagnosis and determination of estrogen and progesterone receptors, and HER2 status. Methacarn and RCL2 fixation preserved DNA integrity as demonstrated by successful amplification and sequencing of large DNA amplicons. Similarly, high-quality RNA could be extracted from methacarn- or RCL2-fixed paraffin-embedded MCF-7 cells, whole breast tumor tissues, or microdissected breast tumor cells, as assessed by electropherogram profiles and real-time reverse transcriptase-polymerase chain reaction quantification of various genes. Moreover, tissue morphology and RNA integrity were preserved after 8 months of storage. Altogether, these results indicate that methacarn, as previously shown, and RCL2, a promising new fixative, have great potential for performing both morphological and molecular analyses on the same fixed tissue sample, even after laser-capture microdissection, and can open new doors for investigating small target lesions such as premalignant breast lesions.
将甲 Carn 固定液和一种新型非交联固定剂 RCL2 与同一浸润性乳腺癌的福尔马林固定或冷冻组织样本进行比较，并评估它们对组织形态学、免疫组织化学以及 DNA 和 RNA 完整性的影响。甲 Carn 固定或 RCL2 固定的石蜡包埋肿瘤的组织形态学与匹配的福尔马林固定组织相似。使用各种抗体进行的免疫组织化学显示，两种固定剂的结果相当，从而能够准确诊断乳腺肿瘤并确定雌激素和孕激素受体以及 HER2 状态。甲 Carn 和 RCL2 固定可保留 DNA 完整性，大型 DNA 扩增子的成功扩增和测序证明了这一点。同样，通过电泳图谱和各种基因的实时逆转录聚合酶链反应定量评估，可从甲 Carn 固定或 RCL2 固定的石蜡包埋 MCF-7 细胞、全乳腺肿瘤组织或显微切割的乳腺肿瘤细胞中提取高质量的 RNA。此外，储存 8 个月后，组织形态学和 RNA 完整性得以保留。总而言之，这些结果表明，如先前所示，甲 Carn 以及一种有前景的新型固定剂 RCL2 在对同一固定组织样本进行形态学和分子分析方面具有巨大潜力，即使在激光捕获显微切割后也是如此，并且可为研究诸如乳腺前期病变等小目标病变开辟新途径。
