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结合细胞类型限制的腺病毒靶向与免疫染色和流式细胞术以鉴定肺癌的起源细胞

Combining Cell Type-Restricted Adenoviral Targeting with Immunostaining and Flow Cytometry to Identify Cells-of-Origin of Lung Cancer.

作者信息

Best Sarah A, Kersbergen Ariena, Asselin-Labat Marie-Liesse, Sutherland Kate D

机构信息

ACRF Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.

Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia.

出版信息

Methods Mol Biol. 2018;1725:15-29. doi: 10.1007/978-1-4939-7568-6_2.

Abstract

Lung cancers display considerable intertumoral heterogeneity, leading to the classification of distinct tumor subtypes. Our understanding of the genetic aberrations that underlie tumor subtypes has been greatly enhanced by recent genomic sequencing studies and state-of-the-art gene targeting technologies, highlighting evidence that distinct lung cancer subtypes may be derived from different "cells-of-origin". Here, we describe the intra-tracheal delivery of cell type-restricted Ad5-Cre viruses into the lungs of adult mice, combined with immunohistochemical and flow cytometry strategies for the detection of lung cancer-initiating cells in vivo.

摘要

肺癌表现出显著的肿瘤间异质性,导致了不同肿瘤亚型的分类。近期的基因组测序研究和先进的基因靶向技术极大地增进了我们对构成肿瘤亚型基础的基因畸变的理解,突显了不同肺癌亚型可能源自不同“起源细胞”的证据。在此,我们描述了将细胞类型受限的Ad5-Cre病毒经气管内递送至成年小鼠肺内,并结合免疫组织化学和流式细胞术策略在体内检测肺癌起始细胞的方法。

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