Abstract：
Fucosyltransferases (Fut) regulate the fucosylation process associated with tumorogenesis in different cancer types. Ascitic fluid (AF) from patients diagnosed with advanced stage of epithelial ovarian cancer (EOC) is considered as a dynamic tumor microenvironment associated with poor prognosis. Previous studies from our laboratory showed increased fucosylation in SKOV-3 and OVCAR-3, cancer-derived cell lines, when these cells were incubated with AFs derived from patients diagnosed with EOC. In the present work we studied three fucosyltransferases (Fut 2, Fut 4, and Fut 8) in SKOV-3, OVCAR-3 and CAOV-3 cell lines in combination with five different AFs from patients diagnosed with this disease, confirming that all tested AFs increased fucosylation. Then, we demonstrate that mRNAs of these three enzymes were overexpressed in the three cell lines under treatment with AFs. SKOV-3 showed the higher overexpression of Fut 2, Fut 4, and Fut 8 in comparison with the control condition. We further confirmed, in the SKOV-3 cell line, by endpoint PCR, WB, and confocal microscopy, that the three enzymes were overexpressed, being Fut 4 the most overexpressed enzyme compared to Fut 2 and Fut 8. These enzymes were concentrated in vesicular structures with a homogeneous distribution pattern throughout the cytoplasm. Moreover, we found that among the three enzymes, only Fut 4 was located inside the nuclei. The nuclear location of Fut 4 was confirmed for the three cell lines. These results allow to propose Fut 2, Fut 4, and Fut 8 as potential targets for EOC treatment or as diagnostic tools for this disease.
摘要：
岩藻糖基转移酶(Fut)调节与不同癌症类型中的肿瘤发生相关的岩藻糖基化过程。诊断为晚期上皮性卵巢癌（EOC）患者的腹水（AF）被认为是与不良预后相关的动态肿瘤微环境。我们实验室的先前研究表明，SKOV-3和OVCAR-3（癌症衍生的细胞系）中的岩藻糖基化增加，当这些细胞与来自诊断为EOC的患者的AF孵育时。在目前的工作中，我们研究了SKOV-3，OVCAR-3和CAOV-3细胞系中的三种岩藻糖基转移酶（Fut2，Fut4和Fut8），并结合了诊断患有这种疾病的患者的五种不同的AF。确认所有测试的AF增加岩藻糖基化。然后，我们证明了这三种酶的mRNA在用AFs处理的三种细胞系中过表达。与对照条件相比，SKOV-3显示Fut2，Fut4和Fut8的过表达更高。我们进一步证实,在SKOV-3细胞系中，通过终点PCR，WB,和共聚焦显微镜，这三种酶过度表达，与Fut2和Fut8相比，Fut4是过表达最多的酶。这些酶集中在囊泡结构中，在整个细胞质中具有均匀的分布模式。此外，我们发现在这三种酶中，只有Fut4位于原子核内部。确认了三种细胞系的Fut4的核位置。这些结果允许提出Fut2、Fut4和Fut8作为EOC治疗的潜在靶标或作为该疾病的诊断工具。
