Abstract
In solid tumors, hypoxia facilitates malignant progression of cancer cells by triggering epithelial-mesenchymal transition (EMT) and cancer stemness. Fascin-1, an actin-bundling protein, takes part in the formation of many actin-based cellular structures. In the present study, we explored the potential functions of hypoxia-induced upregulation of Fascin-1 in liver cancer. Transcriptome RNA-sequencing was conducted to identify hypoxia-related genes. The potential functions of Fascin-1 were evaluated by western blot, transwell migration and invasion assays, sphere-formation assay, tumor xenograft growth, gelatin zymography analysis, immunofluorescence, cell viability assay, soft agar assay, and flow cytometry. We found that Fascin-1 was upregulated by hypoxia in liver cancer cell lines, elevated in liver cancer patients and correlated with larger tumor size, lymph node metastasis, distant metastasis, and shorter overall survival. Knockdown of Fascin-1 suppressed migration, invasion, EMT, stemness, and tumor xenograft growth of liver cancer cells under both normoxia and hypoxia conditions, while forced Fascin-1 expression showed opposite effects. Moreover, hypoxia-induced upregulation of Fascin-1 was regulated by the Akt/Rac1 signaling, and inhibition of Akt/Rac1 signaling by EHop-016 and MK-2206 restrained migration, invasion, EMT, and stemness of liver cancer cells under hypoxia. Furthermore, Fascin-1 knockdown suppressed MMP-2 and MMP-9 expression, impaired actin cytoskeleton rearrangement, inactivated Hippo/YAP signaling, and increased Sorafenib sensitivity in liver cancer cells. Our study provided a novel insight of Fascin-1 in regulating migration, invasion, EMT, and stemness of liver cancer cells under normoxia and hypoxia conditions.
Highlights
Liver cancer is the second leading cause of cancer-related death worldwide and one of few cancers whose incidence and mortality are steadily increasing [1, 2]
Fascin-1 was apparently promotes migration, invasion, epithelial-mesenchymal transition (EMT), and stemness of cancer cells increased in tumor tissues of liver cancer patients, as observed in [5, 19, 20], we evaluated the influence of Fascin-1 knockdown on The Cancer Genome Atlas (TCGA)-hepatocellular carcinoma (HCC) cohort, GSE36376, GSE39791, and GSE102079 (Fig. 1E). these properties of liver cancer cells under normoxia or hypoxia
Several previous studies report that Fascin-1 is elevated in tumor tissues of liver cancer patients, and correlates with larger tumor size, lymph node metastasis, distant metastasis, and poor overall survival [16,17,18]
Summary
Fascin-1 is upregulated by hypoxia in liver cancer cells and because Fascin-1 ranked the top 5 upregulated genes in Hep3B predicts poor prognosis of liver cancer patients cells under hypoxia (Fig. 1A and Supplementary Table 1), and. In qRT-PCR analysis, the expression levels of stemness markers (Oct, Lin, Nanog, and Knockdown of Fascin-1 suppresses migration, invasion, EMT, Sox2) were downregulated by Fascin-1 knockdown in Hep3B stemness, and tumor xenograft growth of liver cancer cells and HuH-6 cells (Fig. 3C). Our results indicated that hypoxia-induced activation of Akt/Rac signaling increased Fascin-1 expression and regulated migration, invasion, EMT, and stemness of liver cancer cells under hypoxia. We found that forced expression of Fascin-1 reduced the phosphorylation of YAP and Lats and increased the total and nuclear levels of YAP in HepG2 cells, indicating that Fascin-1 overexpression promoted Hippo-YAP activation in liver cancer cells (Fig. 7E, F). Our results indicated that knockdown of Fascin increased Sorafenib sensitivity in liver cancer cells
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