目的：探讨miR-4465 靶向高迁移率族蛋白A1（HMGA1）对肝细胞癌 Hep3B 细胞增殖、迁移和侵袭能力的影响。方法：收集2020 年5 月至2021 年9 月在皖南医学院第一附属医院确诊为肝细胞癌患者的16 对癌组织和癌旁组织样本，采用qPCR 分析miR-4465 在肝细胞癌组织和Hep3B、Huh7细胞中的表达情况，双荧光素酶报告基因实验验证miR-4465 与HMGA1的调控关系。按转染物的不同将 Hep3B 细胞分为 mimics-NC 组 、miR-4465-mimics 组、inhibitor-NC 组、miR-4465 inhibitor组、si-NC 组、si-HMGA1 组；另外分组转染mimics-NC+pcDNA-NC、miR-4465 mimics+pcDNA-NC 和miR-4465 mimics+pcDNA-HMGA1进行回复实验。采用qPCR和WB法检测各组细胞中HMGA1 mRNA和蛋白水平的变化，CCK-8法检测各组细胞增殖能力的变化，划痕实验检测各组细胞迁移能力的变化，Transwell 实验检测各组细胞侵袭能力的变化。结果：miR-4465 在肝细胞癌组织和细胞中的表达水平显著低于癌旁组织和正常肝细胞（P<0.05或P<0.001）。转染48 h后，过表达miR-4465 的Hep3B细胞增殖、迁移和侵袭能力均显著下降（P<0.05、P<0.01或P<0.001）；敲低miR-4465 后细胞的增殖、迁移和侵袭能力均明显升高（P<0.05、 P<0.01或P<0.001）。双荧光素酶报告实验验证了HMGA1-3'UTR 与miR-4465 的靶向结合关系，miR-4465 可以靶向下调HMGA1 mRNA 和蛋白质的表达（均P<0.01）。过表达HMGA1 能部分逆转过表达 miR-4465 对细胞增殖、迁移、侵袭的抑制作用及HMGA1 表达的抑制作用（均P<0.05）。结论：miR-4465 通过靶向下调HMGA1 在肝细胞癌Hep3B 细胞中的表达，从而抑制Hep3B细胞的恶性生物学行为。

Objective: To investigate the effect of miR-4465 targeting high mobility group protein A1 (HMGA1) on the proliferation, migration and invasion of hepatocellular carcinoma Hep3B cells. Methods: Sixteen pairs of cancer tissues and adjacent tissue samples from patients diagnosed with hepatocellular carcinoma in the First Affiliated Hospital of Wannan Medical College from May 2020 to September 2021 were collected. The expression of miR-4465 in hepatocellular carcinoma tissues and Hep3B and Huh7 cells was analyzed by qPCR, and the regulatory relationship between miR-4465 and HMGA1 was verified by dual luciferase reporter gene assay. According to the different transfections, Hep3B cells were group, si-NC group, and si-HMGA1 group for transfection; in addition, mimics-NC+pcDNA-NC, miR-4465 mimics+pcDNA-NC, and miR-4465 mimics+pcDNA-HMGA1 were transfected for rescue experiments. The changes of mRNA and protein levels of HMGA1 in each group were detected by qPCR and WB assay; the changes in cell proliferation activity in each group were detected by CCK-8 method; the changes in cell migration ability in each group were detected by scratch assay, and the changes in cell invasion ability in each group were detected by Transwell assay. Results: The expression of miR-4465 in hepatocellular carcinoma tissues and cells was significantly lower than that in adjacent tissues and normal liver cells (P<0.05 or P<0.001). After transfection for 48 h, the proliferation, migration and invasion abilities of Hep3B cells overexpressing miR-4465 were significantly decreased (P<0.05, P<0.01 or P<0.001); the proliferation, migration and invasion abilities of Hep3B cells with miR-4465 knockdown were increased (P<0.05, P<0.01 or P<0.001). The binding relationship between HMGA1-3'UTR and miR-4465 was verified by dual luciferase report gene experiment. miR-4465 could target and down-regulate the mRNA and protein expression of HMGA1 (all P<0.01). Over-expression of HMGA1 partially restored the inhibitory effects of miR-4465 overexpression on cell proliferation, migration and invasion as well as HMGA1 expression (all P<0.05). Conclusion: miR-4465 inhibits the malignant biological behaviors of hepatocellular carcinoma Hep3B cells by down-regulating the expression of HMGA1 in the cells.

国家自然科学基金项目（No. 81772180）；安徽省教育厅省级自然科学重点项目（No. KJ2016A722）；安徽省大学生创新训练项目（No. s202110368042）