miR-17-5p and miR-20a-5p suppress postoperative metastasis of hepatocellular carcinoma via blocking HGF/ERBB3-NF-κB positive feedback loop
miR-17-5p and miR-20a-5p suppress postoperative metastasis of hepatocellular carcinoma via blocking HGF/ERBB3-NF-κB positive feedback loop
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DOI：
10.7150/thno.41365
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Wu, Wei-Zhong
中科院分区：
文献类型：
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作者：
Liu, Dong-Li;Lu, Li-Li;Wu, Wei-Zhong
微小RNA（microRNA，miRNA）表达异常是肝细胞癌（hepatocellular carcinoma，HCC）中常见的事件，但在肝切除术后肝微环境重建过程中，miRNA是旁观者还是参与者，目前尚不清楚。采用Western blot、qRT-PCR、荧光原位杂交、免疫荧光和免疫组化等方法检测miRNA和mRNA在细胞系和患者组织中的表达。在体外和体内研究了其生物学功能。采用染色质免疫沉淀、邻位连接和荧光素酶报告基因分析等方法研究靶基因的特异性结合。结果：肝切除诱导的HGF可促进残余肝癌细胞的转移，且HGF/ERBB 3信号通路的表达与肝切除诱导的肝癌细胞的转移密切相关。miR-17- 5 p和miR-20 a-5 p对肝癌转移有抑制作用。ERBB 3是miR-17- 5 p和miR-20 a-5 p共同作用的靶点。具有较低水平的miR-17- 5 p和miR-20 a-5 p或较高水平的ERBB 3的HCC细胞通常对响应HGF刺激更敏感，并且在体外和体内实验系统中促进转移定植。此外，HGF通过NF-κ B B转录活性在正反馈环中增强ERBB 3表达。特别重要的是，miR-17- 5 p和miR-20 a-5 p水平较低或ERBB 3水平较高的HCC患者手术切除后的OS和PFS生存期显著较短。miR-17- 5 p和miR-20 a-5 p通过阻断HGF/ERBB 3-NF-κ B抑制肝癌术后转移。kappa B正反馈环，为肝癌切除术后转移的预防提供了新的可能策略。
Dysregulation of microRNA (miRNA) is a frequent event in hepatocellular carcinoma (HCC), but little is known whether it is a bystander or an actual player on residual HCC metastasis during liver microenvironment remodeling initiated by hepatectomy.Methods: The differently expressed miRNAs and mRNAs were identified from RNA-seq data. Western blot, qRT-PCR, fluorescence in situ hybridization, immunofluorescence and immunohistochemical were used to detect the expression of miRNA and mRNA in cell lines and patient tissues. The biological functions were investigated in vitro and in vivo. Chromatin immunoprecipitation, proximity ligation and luciferase reporter assay were used to explore the specific binding of target genes. The expression of HGF/ERBB3 signaling was detected by Western blot.Results: In this study, HGF induced by hepatectomy was shown to promote metastasis of residual HCC cells. miR-17-5p and miR-20a-5p were confirmed to play inhibitory roles on HCC metastasis. And ERBB3 was found to be the common target of miR-17-5p and miR-20a-5p. HCC cells with lower levels of miR-17-5p and miR-20a-5p or higher level of ERBB3 were often more sensitive to response HGF stimuli and to facilitate metastatic colonization both in vitro and in vivo experimental systems. Furthermore, HGF reinforced ERBB3 expression by NF-kappa B transcriptional activity in a positive feedback loop. Of particular importance, HCC patients with lower levels of miR-17-5p and miR-20a-5p or higher level of ERBB3 had significantly shorter OS and PFS survivals after surgical resection.Conclusion: miR-17-5p and miR-20a-5p could suppress postoperative metastasis of hepatocellular carcinoma via blocking HGF/ERBB3-NF-kappa B positive feedback loop and offer a new probable strategy for metastasis prevention after HCC resection.