FBW7 increases chemosensitivity in hepatocellular carcinoma cells through suppression of epithelial-mesenchymal transition
FBW7 increases chemosensitivity in hepatocellular carcinoma cells through suppression of epithelial-mesenchymal transition
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DOI：
10.1016/s1499-3872(14)60029-1
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发表时间：
2014-04-15
影响因子：
3.3
通讯作者：
Zheng, Shu-Sen
中科院分区：
文献类型：
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作者：
Yu, Jun;Zhang, Wu;Zheng, Shu-Sen
背景：FBW7 是一种肿瘤抑制因子，调节在细胞分裂、细胞生长和分化中发挥核心作用的蛋白质网络。本研究旨在评估FBW7在不同肝细胞癌（HCC）细胞系中化学敏感性和上皮间质转化（EMT）中的作用，并探讨相关的潜在机制。 方法：培养不同的人HCC细胞系（Hep3B、Huh-7和SNU-449）。通过细胞计数试剂盒8评估细胞活力，并通过实时PCR和Western blotting定量FBW7 mRNA转录和蛋白表达。通过蛋白质印迹和免疫细胞化学评估波形蛋白（间充质生物标志物）和E-钙粘蛋白（上皮生物标志物）的表达。采用Transwell迁移法检测细胞侵袭能力，并使用FBW7质粒或siRNA评估FBW7过表达或沉默对细胞化疗敏感性的影响。结果：FBW7表达影响肿瘤细胞对阿霉素的化疗敏感性以及不同HCC细胞系中肿瘤细胞的侵袭能力。 FBW7(hi)（高 FBW7 表达）Hep3B 和 FBW7(mi)（中位 FBW7 表达）Huh-7 细胞比 FBW7(lo)（低 FBW7 表达）SNU-449 细胞对阿霉素更敏感，侵袭能力更低。 Huh-7和Hep3B细胞中FBW7的沉默诱导了对阿霉素的抵抗并增强了细胞侵袭，而SNU-449细胞中FBW7的过度表达恢复了对阿霉素的敏感性并显着降低了侵袭能力。此外，阿霉素诱导 HCC 细胞向间充质发生 EMT。 Huh-7和Hep3B细胞中FBW7的下调或SNU-449细胞中FBW7的上调改变了EMT的方向。结论：FBW7表达水平影响肿瘤对阿霉素的耐药性和HCC细胞的侵袭能力。因此，FBW7可能通过调节EMT成为HCC化疗的潜在靶点。
BACKGROUND: FBW7 is a tumor suppressor which regulates a network of proteins with central roles in cell division, cell growth and differentiation. This study aimed to evaluate the role of FBW7 in chemosensitivity and epithelial-mesenchymal transition (EMT) in different hepatocellular carcinoma (HCC) cell lines and to investigate the relevant underlying mechanisms.METHODS: Different human HCC cell lines (Hep3B, Huh-7, and SNU-449) were cultured. The cell viability was evaluated by cell counting kit-8, and FBW7 mRNA transcription and protein expression were quantitated by real-time PCR and Western blotting. Expressions of vimentin (mesenchymal biomarker) and E-cadherin (epithelial biomarker) were evaluated by Western blotting and immunocytochemistry. Cell invasion was assayed by Transwell migration, and FBW7 plasmid or siRNA was used to evaluate the effect of FBW7 overexpression or silencing on cell chemosensitivity.RESULTS: FBW7 expression affected tumor cell chemosensitivity to doxorubicin and tumor cell invasive capacity in different HCC cell lines. FBW7(hi) (high FBW7 expression) Hep3B and FBW7(mi) (median FBW7 expression) Huh-7 cells were more sensitive to doxorubicin and lower in invasive capacity than FBW7(lo) (low FBW7 expression) SNU-449 cells. Silencing of FBW7 in Huh-7 and Hep3B cells induced the resistance to doxorubicin and enhanced cell invasion, whereas overexpression of FBW7 in SNU-449 cells restored the sensitivity to doxorubicin and significantly reduced invasive capacity. Furthermore, doxorubicin induced EMT toward mesenchyme in HCC cells. Downregulation of FBW7 in Huh-7 and Hep3B cells or upregulation of FBW7 in SNU-449 cells altered the direction of EMT.CONCLUSIONS: The level of FBW7 expression impacted the tumor resistance to doxorubicin and the invasion capability of HCC cells. FBW7 therefore may be a potential target for the chemotherapy of HCC through the regulation of EMT.