α-1,3-岩藻糖基转移酶IV在肝癌转移中的表达及机制研究

肝脏 ›› 2021, Vol. 26 ›› Issue (10): 1123-1127.

α-1,3-岩藻糖基转移酶IV在肝癌转移中的表达及机制研究

倪磊, 郭健, 于雪, 张凝, 韩依影, 杨瑜爱, 杨建仁, 李春蕊, 贾娟, 刘天华

100029 北京中医药大学中医学院(倪磊,郭健,于雪,张凝,杨建仁,李春蕊,贾娟,刘天华);北京中医药大学第一临床医学院(韩依影,杨瑜爱)

收稿日期:2021-03-20 出版日期:2021-10-31 发布日期:2021-12-07
通讯作者: 刘天华,Email:bucmlth@126.com
基金资助:
北京中医药大学重点攻关项目(2020-JYB-ZDGG-013);国家自然科学基金(81803954)

The expression and mechanism of α-1,3-fucosyltransferase IV in liver cancer metastasis

NI Lei¹, GUO Jian¹, YU Xue¹, ZHANG Ning¹, HAN Yi-ying², YANG Yu-ai², YANG Jian-ren¹, LI Chun-rui¹, JIA Juan¹, LIU Tian-hua¹

1. The School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China;
2. The First Clinical Medical College of Beijing University of Traditional Chinese Medicine, Beijing 100029, China

Received:2021-03-20 Online:2021-10-31 Published:2021-12-07
Contact: LIU Tian-hua,Email:bucmlth@126.com

摘要/Abstract

摘要： 目的探讨α-1,3-岩藻糖基转移酶IV在肝癌转移中的表达及机制。方法通过qRT-PCR检测不同转移潜能肝癌细胞中α-1,3-岩藻糖基转移酶IV(FUT4)的表达水平;利用转化生长因子-β1(Transforming growth factor-β1, TGF-β1)诱导人肝癌MHCC97L细胞构建上皮间质转化(Epithelial-mesenchymal transition, EMT)模型,显微镜下观察细胞形态变化,qRT-PCR及Western blot检测EMT标志分子表达水平,qRT-PCR检测FUT4的表达水平;借助HCMDB数据库获取转移与非转移肝癌样本中FUT4的表达水平及共表达基因,并通过OmicsBean数据库对FUT4及其共表达基因进行KEGG通路富集分析。结果与低转移潜能人肝癌MHCC97L细胞相比,在高转移潜能人肝癌细胞MHCC97H与HCCLM3中FUT4 的mRNA表达水平均显著提高(P＜0.01),分别为MHCC97L细胞中的4.99倍和6.08倍;经TGF-β1诱导的人肝癌MHCC97L细胞呈纺锤形,E-钙黏蛋白的mRNA表达水平显著降低(P<0.001),N-钙黏蛋白、波形蛋白的mRNA表达水平显著提高(P<0.001),蛋白表达变化与mRNA表达变化一致,EMT模型构建成功,且FUT4的mRNA表达水平在该模型中显著提高(P<0.01),为对照组的1.42倍;生物信息学分析结果进一步证实,与非转移肝癌样本相比,转移肝癌样本中FUT4的表达水平显著提高(P<0.01),并且提示FUT4及共表达基因可能通过肿瘤转录调控异常、肌动蛋白细胞骨架调节、ECM受体相互作用等KEGG通路参与肝癌转移过程。结论 FUT4高表达与肝癌细胞EMT及转移密切相关,为肝癌转移的诊断和治疗提供了以FUT4为潜在生物学标志物与靶点的新思路。

关键词: α-1,3岩藻糖基转移酶IV, 肝癌, 转移, 上皮间质转化, 共表达

Abstract: Objective To investigate the expression and mechanism of α-1,3-fucosyltransferase IV in liver cancer metastasis. Methods The expression of FUT4 in liver cancer cell lines exhibiting different metastatic potentials was detected by quantitative Real-time PCR (qRT-PCR). The epithelial-mesenchymal transition (EMT) model was established by stimulating human liver cancer cell line MHCC97L with transforming growth factor-β1 (TGF-β1). Morphologic change was observed by phase-contrast microscope. The expression of mRNAs and proteins of EMT markers were detected by qRT-PCR and Western blot, respectively. The expression of FUT4 was detected by qRT-PCR. The expression levels and co-expression genes of FUT4 in primary liver cancer samples with and without metastasis were investigated using Human Cancer Metastasis Database (HCMDB). Enrichment analysis of KEGG pathways was performed by OmicsBean. Results The mRNA expression levels of FUT4 were significantly upregulated in MHCC97H and HCCLM3, two liver cancer cell lines exhibited high metastatic potential, when compared with liver cancer cell line, MHCC97L, which exhibited low metastatic potential (P<0.01), fold changes were 4.99 and 6.08, respectively. After TGF-β1 treatment, MHCC97L cells were spindle-shaped and the mRNA expression of E-cadherin was significantly reduced (P<0.001), however, the mRNA expressions of N-cadherin and vimentin were significantly increased (P<0.001). The proteins' expression of EMT markers were in parallel as the mRNAs' expression. FUT4 was overexpressed in TGF-β1-induced EMT model in MHCC97L when compared with that of the control (P<0.01), the fold change was 1.42. Bioinformatics analysis results further confirmed that mRNA expression level of FUT4 was significantly upregulated in primary liver cancer samples with metastasis compared with those without metastasis (P<0.01), suggesting that FUT4 might participate in the liver cancer metastasis and programmed by the co-expressing genes belonging to KEGG pathways such as transcriptional mis-regulation in cancer, regulation of actin cytoskeleton, and ECM-receptor interaction. Conclusion These findings not only suggest that high-level expression of FUT4 is closely related to EMT and metastasis of liver cancer, but also provide an insight on using FUT4 as a potential biomarker and target for the diagnosis and treatment of liver cancer metastasis.

Key words: α-1,3-fucosyltransferase IV, liver cancer, metastasis, epithelial-mesenchymal transition, co-expression

倪磊, 郭健, 于雪, 张凝, 韩依影, 杨瑜爱, 杨建仁, 李春蕊, 贾娟, 刘天华. α-1,3-岩藻糖基转移酶IV在肝癌转移中的表达及机制研究[J]. 肝脏, 2021, 26(10): 1123-1127.

NI Lei, GUO Jian, YU Xue, ZHANG Ning, HAN Yi-ying, YANG Yu-ai, YANG Jian-ren, LI Chun-rui, JIA Juan, LIU Tian-hua. The expression and mechanism of α-1,3-fucosyltransferase IV in liver cancer metastasis[J]. Chinese Hepatolgy, 2021, 26(10): 1123-1127.

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