Diffuse large B-cell lymphomas (DLBCLs) are the most common types of Non-Hodgkin's lymphomas and are highly heterogeneous in terms of phenotype and treatment response. The natural course of DLBCLs tumor progression is featured by a flow of events leading to the enhancement of proliferative and invasive capabilities and, therefore, towards the establishment of a more aggressive phenotype. Angiogenesis is a constant hallmark of DLBCLs progression, has prognostic potential and promote DLBCLs dissemination. The study of DLBCLs angiogenesis mechanisms, and the tumor endothelium characterization, will allow us to identify new prognostic/predictive biomarkers to proper patient selection to antiangiogenic treatment. In our previous work, by RNAscope technology, we have demonstrated that Janus kinase (Jak) and signal transducer activator of transcription pathway (STAT) is one of the proangiogenic pathways activated in DLBCLs and it drives neoangiogenesis occurred by vasculogenesis mechanism. Here, we describe a detailed protocol to perform RNAscope technology alone and in combination with immunohistochemistry (called dual RNAscope ISH-IHC) in DLBCLs formalin-fixed, paraffin-embedded sections. We propose dual ISH-IHC as an extremely powerful method to study angiogenesis in DLBCLs, because it allows one to answer important biological questions that are difficult to address using other single methods.

中文翻译：

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RNAscope双重ISH-IHC技术研究弥漫性大B细胞淋巴瘤的血管生成。

弥漫性大B细胞淋巴瘤（DLBCL）是非霍奇金淋巴瘤的最常见类型，在表型和治疗反应方面高度异质。DLBCLs肿瘤进展的自然过程以一系列事件为特征，这些事件导致了增殖和侵袭能力的增强，并因此导致了更具攻击性的表型的建立。血管生成是DLBCLs不断发展的标志，具有预后潜力并促进DLBCLs的传播。的DLBCLs血管生成机理的研究，肿瘤血管内皮细胞特性，将使我们能够确定新的预后/预测生物标记物，以适当的患者选择到抗血管生成治疗。在我们以前的工作中，通过RNAscope技术，我们已经证明Janus激酶（Jak）和转录途径的信号转导激活物（STAT）是DLBCLs中激活的促血管生成途径之一，它驱动由血管生成机制发生的新血管生成。在这里，我们描述了在DLBCLs福尔马林固定，石蜡包埋的切片中单独和与免疫组织化学（称为双重RNAscope ISH-IHC）结合使用RNAscope技术的详细协议。我们提出双重ISH-IHC作为研究DLBCLs中血管生成的一种非常强大的方法，因为它允许人们回答使用其他单一方法难以解决的重要生物学问题。我们描述了在DLBCLs福尔马林固定，石蜡包埋的切片中单独和与免疫组织化学（称为双重RNAscope ISH-IHC）结合使用RNAscope技术的详细协议。我们提出双重ISH-IHC作为研究DLBCLs中血管生成的一种非常有力的方法，因为它允许人们回答使用其他单一方法难以解决的重要生物学问题。我们描述了在DLBCLs福尔马林固定，石蜡包埋的切片中单独和与免疫组织化学（称为双重RNAscope ISH-IHC）结合使用RNAscope技术的详细协议。我们提出双重ISH-IHC作为研究DLBCLs中血管生成的一种非常强大的方法，因为它允许人们回答使用其他单一方法难以解决的重要生物学问题。