Division of Thoracic Surgery, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
STAR Protoc. 2022 May 2;3(2):101367. doi: 10.1016/j.xpro.2022.101367. eCollection 2022 Jun 17.
Here, we describe a protocol for fluorescence-activated cell sorting (FACS) of human EpCAM cells from fresh surgically resected specimens. We then use Q-PCR to identify specific molecular targets associated with the metastatic phenotype. This combined approach enables a qualitative and quantitative gene expression analysis of lung cancer samples. We describe how to use the protocol for Rac GEFs, but it can be applied broadly to other molecular targets. For complete details on the use and execution of this protocol, please refer to Cooke et al. (2021) and Quatromoni et al. (2015).
在这里，我们描述了一种从新鲜手术切除标本中分离人 EpCAM 细胞的荧光激活细胞分选（FACS）的方案。然后，我们使用 Q-PCR 鉴定与转移表型相关的特定分子靶标。这种联合方法能够对肺癌样本进行定性和定量的基因表达分析。我们描述了如何使用 Rac GEFs 方案，但它可以广泛应用于其他分子靶标。有关该方案的使用和执行的完整详细信息，请参阅 Cooke 等人（2021 年）和 Quatromoni 等人（2015 年）。
