[摘要] 目的: 探讨miR-144-3p 通过阻断卷曲受体4（FZD4）/Wnt/β-catenin 信号通路对肝癌Huh-7 细胞增殖、迁移和凋亡的影响及其作用机制。方法：收集2012 年3 月至2017 年7 月在柳州市工人医院手术切除的18 例肝癌患者的癌组织及相应的癌旁组织标本，以及肝癌细胞系Huh-7、SMMC7721 和MHCC97 和人正常肝上皮细胞株THLE-3，用qPCR 检测肝癌组织及细胞系中miR-144-3p 的表达水平。将miR-144-3p mimics/inhibitor 和FZD4 敲降载体转染至Huh-7 细胞中，用CCK-8、Transwell、划痕愈合实验和Annexin V-FITC/PI 染色流式细胞术检测Huh-7 细胞的增殖、迁移和凋亡水平。用双荧光素酶报告基因验证miR-144-3p 和FZD4 的靶向关系。结果：在肝癌组织和细胞系中miR-144-3p 均呈低表达（P<0.05 或P<0.01）。过表达miR-144-3p 可显著抑制Huh-7 细胞增殖、迁移并诱导细胞凋亡（均P<0.01）。双荧光素酶报告基因实验证明miR-144-3p 靶向作用FZD4 并下调其表达水平。体外实验进一步证实，过表达miR-144-3p 靶向FZD4 并阻断Wnt/β-catenin 通路，进而抑制Huh-7 细胞增殖、迁移和促进细胞凋亡（均P<0.01）。结论：miR-144-3p 通过阻断FZD4/Wnt/β-catenin 通路抑制肝癌Huh-7 细胞的恶性生物学行为，为肝癌诊断或治疗提供了潜在分子靶点。

[Abstract] Objective: To investigate the effect of miR-144-3p modulating proliferation, migration and apoptosis of liver cancer Huh-7 cells through blocking frizzled class receptor 4 (FZD4)/Wnt/β -catenin pathway and the possible mechanism. Methods: A total of 18 pairs of cancer tissues and corresponding para-cancerous tissues from liver cancer patients, who underwent surgery in Workers' Hospital of Liuzhou City from March 2012 to July 2017, were collected for this study; in addition, hepatic cancer cell lines (Huh-7, SMMC7721 and MHCC97) and human normal liver epithelial cell line THLE-3 were also collected. The expression of miR-144-3p in liver cancer tissues and cell lines was detected by qPCR. MiR-144-3p mimics/inhibitor and FZD4 siRNA were transfected into liver cancer Huh-7 cells; the proliferation, migration and apoptosis of Huh-7 cells were evaluated by CCK-8 assay, Transwell assay, wound healing assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. The interaction between miR-144-3p and FZD4 was verified by dual-luciferase reporter gene assay. Results: The expression of miR-144-3p was down-regulated in liver cancer tissues and cell lines (P<0.05 or P<0.01). Over-expression of miR-144-3p significantly inhibited cell proliferation viability, migration but induced apoptosis of Huh-7 cells (all P<0.01). Moreover, dual-luciferase reporter gene assay showed that miR-144-3p directly interacted with FZD4 and suppressed its expression. Furthermore, in vitro experiments verified that miR-144-3p targeted FZD4 to suppress the proliferation,migration and promote apoptosis of Huh-7 cells via blocking Wnt/β -catenin pathway (all P<0.01). Conclusion: miR-144-3p inhibits malignant biological behaviors of liver cancer Huh-7 cells via blocking Wnt/FZD4/β-catenin signaling pathway, which may provide potential molecular targets for early diagnosis or treatment of liver cancer.

柳州市工人医院科研资助项目（No. 2017BH20305, No. Z20170896）