Real-Time PCR Assay for the Analysis of Alternative Splicing of Immune Mediators in Cancer.
Real-Time PCR Assay for the Analysis of Alternative Splicing of Immune Mediators in Cancer.
复制标题
DOI：
10.1007/978-1-0716-0247-8_21
复制
发表时间：
2020
影响因子：
--
通讯作者：
Ruizhi Wang;M. F. Hossain;Jovan Mirkovic;Samuel Sabzanov;M. Ruggiu
中科院分区：
文献类型：
--
作者：
Ruizhi Wang;M. F. Hossain;Jovan Mirkovic;Samuel Sabzanov;M. Ruggiu
选择性剪接是一种非常有效的产生蛋白质组多样性和从有限数量的基因调节细胞稳态的方法。此外，来自相同前mrna的不同剪接变体的相对数量的变化是癌症的一个标志，而选择性剪接mrna的异常表达与癌症的发生和进展有关。因此，剪接变异体是评估疾病进展和临床预后的重要工具，并有望成为治疗干预的潜在靶点。为了了解这些剪接变异体在癌症中的作用，无论在正常状态还是在疾病状态下，能够准确量化它们在不同细胞类型和器官中的表达水平是至关重要的。在本章中，我们描述了一种通过定量PCR有效检测、分析和量化癌症中趋化因子、细胞因子及其受体和配体等免疫介质的选择性剪接模式的方案。
Alternative splicing evolved as a very efficient way to generate proteome diversity and to regulate cell homeostasis from a limited number of genes. Moreover, changes in the relative amounts of different splice variants derived from the same pre-mRNA are a hallmark in cancer, and aberrant expression of alternatively spliced mRNAs has been linked to cancer initiation and progression. Therefore, splice variants are critical tools to assess disease progression and clinical prognosis, and hold great promise as potential targets for therapeutic intervention. In order to understand the role that such splice variants play in cancer, it is vital to be able to accurately quantify their expression levels in different cell types and organs, both in normal conditions and in disease. In this chapter we describe a protocol to efficiently detect, analyze, and quantify alternative splicing patterns of immune mediators such as chemokines, cytokine and their receptors and ligands in cancer by quantitative PCR.