Institute of Immunology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany.
Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany.
BMC Cancer. 2020 May 19;20(1):446. doi: 10.1186/s12885-020-06952-9.
Several physiological (fertilization, placentation, wound healing) and pathophysiological processes (infection with enveloped viruses, cancer) depend on cell fusion. In cancer it was postulated that the fusion of cancer cells with normal cells such as macrophages or stem cells may not only give rise to hybrid cells exhibiting novel properties, such as an increased metastatic capacity and drug resistance, but possibly also cancer stem/ initiating cell properties. Hence, hybrid clone cells (M13HS, M13MDA435 and M13MDA231) that were derived from spontaneous fusion events of human M13SV1-EGFP-Neo breast epithelial cells and HS578T-Hyg, MDA-MB-435-Hyg and MDA-MB-231-Hyg cancer cells were investigated regarding potential in vitro cancer stem/ initiating cell properties.
CD44/CD24 expression pattern and ALDH1 activity of parental cells and hybrid clones was determined by flow cytometry. A colony formation and mammosphere formation assay was applied to determine the cells' capability to form colonies and mammospheres. Sox9, Slug and Snail expression levels were determined by Western blot analysis.
Flow cytometry revealed that all hybrid clone cells were CD44/CD24, but differed markedly among each other regarding ALDH1 activity. Likewise, each hybrid clone possessed a unique colony formation and mammosphere capacity as well as unique Snail, Slug and Sox9 expression patterns. Nonetheless, comparison of hybrid clones revealed that M13HS hybrids exhibited more in vitro cancer stem/ initiating cell properties than M13MDA231 and M13MDA435 hybrids, such as more ALDH1 positive cells or an increased capacity to form colonies and mammospheres.
The fate whether cancer stem/ initiating cells may originate from cell fusion events likely depends on the specific characteristics of the parental cells.
几种生理（受精、胎盘形成、伤口愈合）和病理生理过程（包膜病毒感染、癌症）依赖于细胞融合。在癌症中，有人假设癌细胞与正常细胞（如巨噬细胞或干细胞）融合不仅会产生具有新特性的杂交细胞，例如增加转移能力和耐药性，还可能具有癌症干细胞/起始细胞特性。因此，研究了源自人 M13SV1-EGFP-Neo 乳腺上皮细胞和 HS578T-Hyg、MDA-MB-435-Hyg 和 MDA-MB-231-Hyg 癌细胞自发融合事件的杂交克隆细胞（M13HS、M13MDA435 和 M13MDA231），以研究其潜在的体外癌症干细胞/起始细胞特性。
通过流式细胞术测定亲本细胞和杂交克隆细胞的 CD44/CD24 表达模式和 ALDH1 活性。应用集落形成和类器官形成实验来确定细胞形成集落和类器官的能力。通过 Western blot 分析测定 Sox9、Slug 和 Snail 的表达水平。
流式细胞术显示，所有杂交克隆细胞均为 CD44/CD24，但 ALDH1 活性在彼此之间差异显著。同样，每个杂交克隆细胞均具有独特的集落形成和类器官形成能力以及独特的 Snail、Slug 和 Sox9 表达模式。尽管如此，对杂交克隆细胞的比较表明，M13HS 杂交细胞比 M13MDA231 和 M13MDA435 杂交细胞表现出更多的体外癌症干细胞/起始细胞特性，例如更多的 ALDH1 阳性细胞或形成集落和类器官的能力增加。
癌症干细胞/起始细胞是否可能源自细胞融合事件，可能取决于亲本细胞的特定特征。
