Abstract:
Cell fusion is a biological process that is crucial for the development and homeostasis of different tissues, but it is also pathophysiologically associated with tumor progression and malignancy. The investigation of cell fusion processes is difficult because there is no standardized marker. Many studies therefore use different systems to observe and quantify cell fusion in vitro and in vivo. The comparability of the results must be critically questioned, because both the experimental procedure and the assays differ between studies. The comparability of the fluorescence-based fluorescence double reporter (FDR) and dual split protein (DSP) assay was investigated as part of this study, in which general conditions were kept largely constant. In order to be able to induce both a high and a low cell fusion rate, M13SV1 breast epithelial cells were modified with regard to the expression level of the fusogenic protein Syncytin-1 and its receptor ASCT2 and were co-cultivated for 72 h with different breast cancer cell lines. A high number of fused cells was found in co-cultures with Syncytin-1-overexpressing M13SV1 cells, but differences between the assays were also observed. This shows that the quantification of cell fusion events in particular is highly dependent on the assay selected, but the influence of fusogenic proteins can be visualized very well.
摘要:
细胞融合是一个生物学过程,对不同组织的发育和稳态至关重要,但它在病理生理上也与肿瘤进展和恶性肿瘤有关。细胞融合过程的研究很困难,因为没有标准化的标记。因此,许多研究使用不同的系统来观察和定量体外和体内的细胞融合。结果的可比性必须受到严格质疑,因为实验程序和试验方法在不同的研究之间是不同的。作为本研究的一部分,研究了基于荧光的荧光双报告(FDR)和双分裂蛋白(DSP)测定的可比性,一般情况基本保持不变。为了能够诱导高和低的细胞融合率,M13SV1乳腺上皮细胞在融合蛋白Syncytin-1及其受体ASCT2的表达水平方面进行了修饰,并与不同的乳腺癌细胞系共培养72小时。在与Syncytin-1过表达M13SV1细胞的共培养物中发现了大量的融合细胞,但也观察到试验之间的差异。这表明细胞融合事件的定量尤其高度依赖于所选择的测定。但是融合蛋白的影响可以很好地可视化。