BACKGROUND Adhesion molecules distributed on the cell-surface depends upon their dynamic trafficking that plays an important role during cancer progression. ADP-ribosylation factor 6 (Arf6) is a master regulator of membrane trafficking. CD147, a tumor-related adhesive protein, can promote the invasion of liver cancer. However, the role of Arf6 in CD147 trafficking and its contribution to liver cancer progression remain unclear. METHODS Stable liver cancer cell lines with Arf6 silencing and over-expression were established. Confocal imaging, flow cytometry, biotinylation and endomembrane isolation were used to detect CD147 uptake and recycling. GST-pull down, gelatin zymography, immunofluorescence, cell adhesion, aggregation and tight junction formation, Transwell migration, and invasion assays were used to examine the cellular phenotypes. GEPIA bioinformatics, patient's specimens and electronic records collection, and immunohistochemistry were performed to obtain the clinical relevance for Arf6-CD147 signaling. RESULTS We found that the endocytic recycling of CD147 in liver cancer cells was controlled by Arf6 through concurrent Rab5 and Rab22 activation. Disruption of Arf6-mediated CD147 trafficking reduced the cell-matrix and cell-cell adhesion, weakened cell aggregation and junction stability, attenuated MMPs secretion and cytoskeleton reorganization, impaired HGF-stimulated Rac1 activation, and markedly decreased the migration and invasion of liver cancer cells. Moreover, high-expression of the Arf6-CD147 signaling components in HCC (hepatocellular carcinoma) was closely correlated with poor clinical outcome of patients. CONCLUSIONS Our results revealed that Arf6-mediated CD147 endocytic recycling is required for the malignant phenotypes of liver cancer. The Arf6-driven signaling machinery provides excellent biomarkers or therapeutic targets for the prevention of liver cancer.

中文翻译：

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Arf6驱动的CD147内吞再循环决定了HCC恶性表型。

背景技术分布在细胞表面上的粘附分子取决于它们的动态运输，该动态运输在癌症进展期间起重要作用。ADP-核糖基化因子6（Arf6）是膜运输的主要调节剂。CD147是一种与肿瘤相关的粘附蛋白，可以促进肝癌的侵袭。然而，Arf6在CD147贩运中的作用及其对肝癌进展的作用仍不清楚。方法建立稳定的具有Arf6沉默和过度表达的肝癌细胞系。共聚焦成像，流式细胞仪，生物素化和膜隔离被用来检测CD147的摄取和回收。GST下拉，明胶酶谱，免疫荧光，细胞粘附，聚集和紧密连接形成，Transwell迁移和侵袭实验被用来检查细胞表型。进行了GEPIA生物信息学，患者标本和电子记录收集以及免疫组织化学分析，以获取Arf6-CD147信号传导的临床相关性。结果我们发现肝癌细胞中CD147的内吞再循环受Arf6通过同时的Rab5和Rab22激活来控制。Arf6介导的CD147贩运的中断减少了细胞基质和细胞间粘附，减弱了细胞聚集和连接稳定性，减弱了MMP分泌和细胞骨架重组，削弱了HGF刺激的Rac1活化，并显着降低了肝癌细胞的迁移和侵袭。 。此外，肝癌（肝细胞癌）中Arf6-CD147信号转导成分的高表达与患者不良的临床预后密切相关。结论我们的结果表明，肝癌的恶性表型需要Arf6介导的CD147内吞再循环。Arf6驱动的信号传导机制为预防肝癌提供了出色的生物标志物或治疗靶标。