Exosomes secreted by chronic hepatitis B patients with PNALT and liver inflammation grade ≥ A2 promoted the progression of liver cancer by transferring miR-25-3p to inhibit the co-expression of TCF21 and HHIP
Exosomes secreted by chronic hepatitis B patients with PNALT and liver inflammation grade ≥ A2 promoted the progression of liver cancer by transferring miR-25-3p to inhibit the co-expression of TCF21 and HHIP
复制标题
PNALT且肝脏炎症等级≥A2级的慢性乙型肝炎患者分泌的外泌体通过转染miR-25-3p抑制TCF21和HHIP的共表达促进肝癌进展
DOI：
10.1111/cpr.12833
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发表时间：
2020-06-11
影响因子：
8.5
通讯作者：
Fu, Xiaoyu
中科院分区：
文献类型：
--
作者：
Ouyang, Yi;Tang, Yujing;Fu, Xiaoyu
目的探讨慢性乙型病毒性肝炎(CHB)合并PNALT和肝脏炎症分级(>；=A2)患者分泌的外切体影响肝癌发生发展的机制。材料与方法采用RT-PCR、Western blotting和免疫组织化学方法检测基因表达。采用CCK-8、集落形成、Transwell、划痕和流式细胞仪检测细胞活力、增殖、凋亡和转移能力。免疫共沉淀法检测TCF21与HHIP的相互作用。用荧光素酶报告基因检测TCF21/HHIP与miR-25-3P的结合。裸鼠移植瘤研究表明miR-25-3P具有肿瘤生长能力。使用TargetScan、EVmiRNA、TCGA、GEO、David、COEXPEDIA、UALCAN、UCSC和人类蛋白质图谱数据库进行生物信息学分析。结果CHB-PNALT-Exo(>；=A2)可促进HepG2.2.15细胞的增殖和转移。MIR-25-3p在CHB-PNALT-Exo(>；=A2)中上调。MIR-25-3p过表达促进细胞增殖和转移，并与CHB-PNALT(>；=A2)患者的生存不良有关。MiR-25-3p抑制剂可阻断CHB-PNALT-Exo(>；=A2)的促增殖和促转移作用。TCF21与HHIP直接相互作用。抑制TCF21或HHIP可促进细胞增殖和转移。敲除TCF21或HHIP可拮抗含miR-25-3P抑制剂的CHB-PNALT-Exo(>；=A2)对细胞增殖、转移及Ki67、E-钙粘蛋白和caspase-3/-9表达的影响。结论CHB-PNALT-Exo转导miR-25-3p可抑制TCF21和HHIP的共表达，从而促进肝癌的发生发展。
Objectives The current study aimed to investigate the mechanism by which exosomes secreted by CHB patients with PNALT and liver inflammation grade (>= A2) affected the development of liver cancer. Materials and methods Gene expression was assessed by RT-PCR, Western blotting and immunohistochemistry. CCK-8, colony formation, transwell, scratch-wound and flow cytometry assays were used to detect cell viability, proliferation, apoptosis and metastasis. The interaction of TCF21 and HHIP was assessed by co-immunoprecipitation assay. Luciferase reporter was used to detect the combination of TCF21/HHIP and miR-25-3p. Xenograft studies in nude mice manifested tumour growth ability of miR-25-3p. Bioinformatics analyses were conducted using TargetScan, EVmiRNA, TCGA, GEO, DAVID,COEXPEDIA,UALCAN, UCSC and the Human Protein Atlas databases. Results CHB-PNALT-Exo (>= A2) promoted the proliferation and metastasis of HepG2.2.15 cells. miR-25-3p was upregulated in CHB-PNALT-Exo (>= A2). miR-25-3p overexpression promoted cell proliferation and metastasis and was related to poor survival in patients with CHB-PNALT (>= A2). The cell proliferation- and metastasis-promoting functions of CHB-PNALT-Exo (>= A2) were abolished by miR-25-3p inhibitors. TCF21 directly interacted with HHIP. Inhibition of TCF21 or HHIP promoted cell proliferation and metastasis. Knockdown of TCF21 or HHIP counteracted the effects of CHB-PNALT-Exo (>= A2) containing miR-25-3p inhibitor on cell proliferation, metastasis and the expression of Ki67, E-cadherin and caspase-3/-9. Conclusions Transfer of miR-25-3p by CHB-PNALT-Exo promoted the development of liver cancer by inhibiting the co-expression of TCF21 and HHIP.