Fluorescence-based codetection with protein markers reveals distinct cellular compartments for altered MicroRNA expression in solid tumors.
Fluorescence-based codetection with protein markers reveals distinct cellular compartments for altered MicroRNA expression in solid tumors.
复制标题
DOI：
10.1158/1078-0432.ccr-10-1152
复制
发表时间：
2010-08-15
期刊：
影响因子：
--
通讯作者：
Korc M
中科院分区：
文献类型：
--
作者：
Sempere LF;Preis M;Yezefski T;Ouyang H;Suriawinata AA;Silahtaroglu A;Conejo-Garcia JR;Kauppinen S;Wells W;Korc M
高通量分析实验已将 microRNA (miRNA) 表达的改变与不同类型的癌症联系起来。肿瘤组织是一种异质混合物，不仅包含癌细胞，还包含支持性和反应性肿瘤微环境元素。为了阐明实体瘤中 miRNA 表达改变的临床意义，我们开发了一种灵敏的基于荧光的原位杂交 (ISH) 方法，用于可视化福尔马林固定、石蜡包埋的组织标本中单个细胞内 miRNA 的积累。该 ISH 方法的实施与常规临床免疫组织化学 (IHC) 检测兼容，能够在同一组织切片中检测 miRNA 和蛋白质标记物，以进行共定位和功能研究。我们使用这种组合的 ISH/IHC 检测来研究一组癌症相关 miRNA，包括在乳腺癌、结直肠癌、肺癌、胰腺癌和前列腺癌中经常检测到低水平（miR-34a 和 miR-126）和高水平（miR-21 和 miR-155）的 miRNA。尽管每种特定癌症类型都有不同的组织病理学改变，但总的趋势表明 miRNA 表达改变的不同来源细胞。尽管 miR-21 和 miR-34a 的表达改变在癌细胞内表现出来，但 miR-126 和 miR-155 的表达改变主要分别局限于内皮细胞和免疫细胞。这些结果表明 miRNA 通过内在地影响癌细胞生物学或通过调节基质、血管和免疫反应来异质性地参与癌发生。我们描述了一种快速、灵敏的多色 ISH/IHC 检测，并表明它可以作为一种研究工具广泛应用，以更好地了解癌症中 miRNA 表达改变的病因相关性。
High-throughput profiling experiments have linked altered expression of microRNAs (miRNA) to different types of cancer. Tumor tissues are a heterogeneous mixture of not only cancer cells, but also supportive and reactive tumor microenvironment elements. To clarify the clinical significance of altered miRNA expression in solid tumors, we developed a sensitive fluorescence-based in situ hybridization (ISH) method to visualize miRNA accumulation within individual cells in formalin-fixed, paraffin-embedded tissue specimens. This ISH method was implemented to be compatible with routine clinical immunohistochemical (IHC) assays to enable the detection of miRNAs and protein markers in the same tissue section for colocalization and functional studies. We used this combined ISH/IHC assay to study a subset of cancer-associated miRNAs, including miRNAs frequently detected at low (miR-34a and miR-126) and high (miR-21 and miR-155) levels, in a panel of breast, colorectal, lung, pancreas, and prostate carcinomas. Despite the distinct histopathologic alterations of each particular cancer type, general trends emerged that pinpointed distinct source cells of altered miRNA expression. Although altered expressions of miR-21 and miR-34a were manifested within cancer cells, those of miR-126 and miR-155 were predominantly confined to endothelial cells and immune cells, respectively. These results suggest a heterogeneous participation of miRNAs in carcinogenesis by intrinsically affecting cancer cell biology or by modulating stromal, vascular, and immune responses. We described a rapid and sensitive multicolor ISH/IHC assay and showed that it could be broadly applied as an investigational tool to better understand the etiologic relevance of altered miRNA expression in cancer.