Airway brushing as a new experimental methodology to detect airway gene expression signatures in mouse lung squamous cell carcinoma.
Airway brushing as a new experimental methodology to detect airway gene expression signatures in mouse lung squamous cell carcinoma.
复制标题
DOI：
10.1038/s41598-018-26902-7
复制
发表时间：
2018-06-11
影响因子：
4.6
通讯作者：
You M
中科院分区：
文献类型：
--
作者：
Pan J;Xiong D;Zhang Q;Szabo E;Miller MS;Lubet RA;Wang Y;You M
由于暴露于环境毒物中，肺部会发生“野区癌变”效应，导致在含有癌基因或肿瘤抑制基因突变的受损上皮内出现一场初始但形态正常的细胞。与人类不同的是，人类的气道损伤与肺癌相关的领域长期以来一直是通过支气管镜检查获得的气道刷来研究的，由于小鼠气道的尺寸较小，没有方法可用于小鼠的类似研究。在本方案中，我们描述了一种详细的方法，从一只活老鼠进行气道刷洗，它可以从同一只老鼠重复采样，从而模仿人类使用的支气管镜检查方案。在n -亚硝基-三氯乙基脲（NTCU）诱导的小鼠肺鳞状细胞癌（SCC）模型中，我们分离了具有完整细胞膜结构的气道上皮细胞，然后进行转录组测序（RNA-Seq）。我们发现PI3K信号网络的激活在瘤前SCC病变小鼠的细胞学正常支气管气道上皮细胞中最为显著。长时间暴露于NTCU也诱导了NF-kappaB的激活（NFƙB）， PI3K的下游途径；这种ntcu诱导的肺SCC进展可以通过阻断NFƙB途径逆转。
As a consequence of exposure to environmental toxicants, a “field cancerization” effect occurs in the lung, resulting in the development of a field of initiated, but morphologically normal appearing cells within a damaged epithelium containing mutations in oncogene or tumor suppressor genes. Unlike humans, whose airway field of injury associated with lung cancer has long been investigated with airway brushings obtained via bronchoscopy, no methods are available for similar studies in the mouse due to the small size of the murine airways. In this protocol, we describe a detailed method for performing airway brushing from a live mouse, which enables repeated sampling from the same mouse and thus, mimicking the bronchoscopy protocol used in humans. Using this approach in the N-nitroso-tris-chloroethylurea (NTCU)-induced mouse lung squamous cell carcinoma (SCC) model, we isolated airway epithelial cells with intact cell membrane structure and then performed transcriptome sequencing (RNA-Seq). We found activation of the PI3K signaling network to be the most significant in cytologically normal bronchial airway epithelial cells of mice with preneoplastic lung SCC lesions. Prolonged exposure to NTCU also induced activation of NF-kappaB (NFƙB), the downstream pathway of PI3K; this NTCU-induced lung SCC progression can be reversed by blocking the NFƙB pathway.
登录
查看更多内容
DOI：
10.1158/1940-6207.capr-15-0305
发表时间：
2016-01
期刊：
Cancer prevention research (Philadelphia, Pa.)
影响因子：
--
作者：
Merrick DT;Gao D;Miller YE;Keith RL;Baron AE;Feser W;Kennedy TC;Blatchford PJ;Braudrick S;Hirsch FR;Heasley L;Bunn PA Jr;Franklin WA
通讯作者：
Franklin WA
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
14.9
作者：
Ritchie ME;Phipson B;Wu D;Hu Y;Law CW;Shi W;Smyth GK
通讯作者：
Smyth GK
影响因子：
--
作者：
Kellar A;Egan C;Morris D
通讯作者：
Morris D
影响因子：
14.8
作者：
Krupnick, Alexander S.;Lin, Xue;Li, Wenjun;Okazaki, Mikio;Lai, Jiaming;Sugimoto, Seiichiro;Richardson, Steven B.;Kornfeld, Christopher G.;Garbow, Joel R.;Patterson, G. Alexander;Gelman, Andrew E.;Kreisel, Daniel
通讯作者：
Kreisel, Daniel