a Department of Biochemistry and Molecular Biology , Gainesville , FL.
b Department of Medicine , University of Florida , Gainesville , FL.
Cancer Biol Ther. 2018 Jul 3;19(7):598-608. doi: 10.1080/15384047.2018.1449612. Epub 2018 Apr 13.
Human cell lines are an important resource for research, and are often used as in vitro models of human diseases. In response to the mandate that all cells should be authenticated, we discovered that the MDA-MB-231 cells that were in use in our lab, did not validate based on the alleles of 9 different markers (STR Profile). We had been using this line as a model of triple negative breast cancer (TNBC) that has the ability to form tumors in immuno-compromised mice. Based on marker analysis, these cells most closely resembled the MCF10A line, which are a near diploid and normal mammary epithelial line. Yet, the original cells express carbonic anhydrase IX (CAIX) both constitutively and in response to hypoxia and are features that likely drive the aggressive nature of these cells. Thus, we sought to sub-purify CAIX-expressing cells using Fluorescence Activated Cell Sorting (FACS). These studies have revealed a new line of cells that we have name UFH-001, which have the TNBC phenotype, are positive for CAIX expression, both constitutively and in response to hypoxia, and behave aggressively in vivo. These cells may be useful for exploring mechanisms that underlie progression, migration, and metastasis of this phenotype. In addition, constitutive expression of CAIX allows its evaluation as a therapeutic target, both in vivo and in vitro.
人源细胞系是研究的重要资源,通常被用作人类疾病的体外模型。为了响应所有细胞都应经过验证的要求,我们发现实验室中使用的 MDA-MB-231 细胞在 9 个不同标记物(STR 谱)的等位基因上无法验证。我们一直将该细胞系用作三阴性乳腺癌(TNBC)的模型,该模型具有在免疫功能低下的小鼠中形成肿瘤的能力。根据标记物分析,这些细胞最接近 MCF10A 系,它们是近二倍体和正常乳腺上皮细胞系。然而,原始细胞持续表达碳酸酐酶 IX(CAIX),并对缺氧有反应,这些特征可能驱动这些细胞的侵袭性。因此,我们试图使用荧光激活细胞分选(FACS)对 CAIX 表达细胞进行亚纯化。这些研究揭示了一种新的细胞系,我们将其命名为 UFH-001,该细胞系具有 TNBC 表型,CAIX 表达阳性,持续表达且对缺氧有反应,并在体内表现出侵袭性。这些细胞可能有助于探索这种表型进展、迁移和转移的机制。此外,CAIX 的组成型表达允许其作为治疗靶点进行体内和体外评估。