Drews-Elger Katherine, Brinkman Joeli A, Miller Philip, Shah Sanket H, Harrell J Chuck, da Silva Thiago G, Ao Zheng, Schlater Amy, Azzam Diana J, Diehl Kathleen, Thomas Dafydd, Slingerland Joyce M, Perou Charles M, Lippman Marc E, El-Ashry Dorraya
Sylvester Comprehensive Cancer Center, Department of Medicine, University of Miami Miller School of Medicine, Biomedical Research Building, 1501 NW 10th Avenue, Miami, FL, 33136, USA.
Breast Cancer Res Treat. 2014 Apr;144(3):503-17. doi: 10.1007/s10549-014-2887-9. Epub 2014 Feb 25.
Our goal was to establish primary cultures from dissociation of breast tumors in order to provide cellular models that may better recapitulate breast cancer pathogenesis and the metastatic process. Here, we report the characterization of six cellular models derived from the dissociation of primary breast tumor specimens, referred to as "dissociated tumor (DT) cells." In vitro, DT cells were characterized by proliferation assays, colony formation assays, protein, and gene expression profiling, including PAM50 predictor analysis. In vivo, tumorigenic and metastatic potential of DT cultures was assessed in NOD/SCID and NSG mice. These cellular models differ from recently developed patient-derived xenograft models in that they can be used for both in vitro and in vivo studies. PAM50 predictor analysis showed DT cultures similar to their paired primary tumor and as belonging to the basal and Her2-enriched subtypes. In vivo, three DT cultures are tumorigenic in NOD/SCID and NSG mice, and one of these is metastatic to lymph nodes and lung after orthotopic inoculation into the mammary fat pad, without excision of the primary tumor. Three DT cultures comprised of cancer-associated fibroblasts (CAFs) were isolated from luminal A, Her2-enriched, and basal primary tumors. Among the DT cells are those that are tumorigenic and metastatic in immunosuppressed mice, offering novel cellular models of ER-negative breast cancer subtypes. A group of CAFs provide tumor subtype-specific components of the tumor microenvironment (TME). Altogether, these DT cultures provide closer-to-primary cellular models for the study of breast cancer pathogenesis, metastasis, and TME.
我们的目标是通过解离乳腺肿瘤来建立原代培养物，以提供可能更好地概括乳腺癌发病机制和转移过程的细胞模型。在此，我们报告了从原发性乳腺肿瘤标本解离获得的六种细胞模型的特征，这些模型被称为“解离肿瘤（DT）细胞”。在体外，通过增殖测定、集落形成测定、蛋白质和基因表达谱分析（包括PAM50预测分析）对DT细胞进行了表征。在体内，在NOD/SCID和NSG小鼠中评估了DT培养物的致瘤性和转移潜力。这些细胞模型与最近开发的患者来源的异种移植模型不同，因为它们可用于体外和体内研究。PAM50预测分析表明，DT培养物与其配对的原发性肿瘤相似，属于基底型和Her2富集型亚型。在体内，三种DT培养物在NOD/SCID和NSG小鼠中具有致瘤性，其中一种在原位接种到乳腺脂肪垫后转移至淋巴结和肺，且未切除原发性肿瘤。从腔面A型、Her2富集型和基底型原发性肿瘤中分离出三种由癌症相关成纤维细胞（CAF）组成的DT培养物。在DT细胞中，有一些在免疫抑制小鼠中具有致瘤性和转移性，为ER阴性乳腺癌亚型提供了新的细胞模型。一组CAF提供了肿瘤微环境（TME）的肿瘤亚型特异性成分。总之，这些DT培养物为研究乳腺癌发病机制、转移和TME提供了更接近原代的细胞模型。
