目的：探讨miR-3612 靶向信号素（SEMA）4C对肝细胞癌细胞增殖与侵袭能力的影响。方法: 收集2020 年5 月至2021 年9月间在皖南医学院第一附属医院弋矶山医院手术切除的肝细胞肝癌的40对癌组织和相应癌旁组织，常规培养肝细胞癌Hep3B和Huh7细胞，将其分为对照组、miR-3612 mimics-NC 组、miR-3612 mimics 组、miR-3612 inhibitor-NC组、miR-3612 inhibitor组、si-NC 组、si-SEMA4C 组、mimics-NC+pcDNA-NC 组、miR-3612 mimics+pcDNA-NC 组和miR-3612 mimics+pcDNA-SEMA4C组，用转染试剂将相应的核酸和质粒转染各组细胞。qPCR 法检测miR-3612 和SEMA4C mRNA 在肝细胞癌组织和Hep3B 和Huh7细胞中的表达，双荧光素酶报告基因实验和免疫共沉淀（RIP）实验验证miR-3612 与SEMA4C的结合及调控关系，qPCR 法和WB法检测转染后各组Hep3B和Huh7细胞中miR-3612、SEMA4C mRNA和蛋白的表达，CCK-8法、细胞划痕实验和Transwell小室实验分别检测各组Hep3B和Huh7细胞的增殖、迁移和侵袭能力。结果: miR-3612 在肝细胞癌组织和Hep3B和Huh7细胞中呈低表达（P<0.001），而SEMA4C 则呈高表达（P<0.001），过表达miR-3612 可抑制Hep3B 和Huh7 细胞的增殖、迁移、侵袭和vimentin、SEMA4C蛋白的表达，促进E-cadherin 蛋白的表达（P<0.05 或P<0.01 或P<0.001），敲低miR-3612 则促进Hep3B和Huh7细胞的增殖、迁移、侵袭和SEMA4C蛋白的表达（P<0.05 或P<0.01 或P<0.001）。双荧光素酶报告基因实验和RIP 实验证实miR-3612 与SEMA4C可直接结合（P<0.001），miR-3612 与SEMA4C的表达呈负相关也间接证明了这一点（P<0.001）。敲减SEMA4C能明显抑制Hep3B、Huh7细胞的增殖、侵袭和迁移能力（P<0.05 或P<0.01 或P<0.001），过表达SEMA4C可逆转过表达miR-3612对Hep3B和Huh7 细胞增殖、迁移、侵袭和EMT的抑制作用（P<0.05 或P<0.01 或P<0.001）。结论: miR-3612 通过调控SEMA4C表达影响Hep3B和Huh7细胞的恶性生物学行为，miR-3612有望成为临床肝细胞癌治疗的潜在靶点。

Objective: To investigate the effect of miR-3612 targeting semaphorin (SEMA)4C on the proliferation and invasion ability of hepatocellular carcinoma cells. Methods: Forty pairs of cancerous tissues and corresponding paracancerous tissues of hepatocellular carcinoma that surgically resected at Yijishan Hospital, the First Affiliated Hospital of Wannan Medical College between May 2020 and September 2021 were collected for this study. Hepatocellular carcinoma Hep3B and Huh7 cells were routinely cultured and divided into control group, miR-3612 mimics-NC group, miR-3612 mimics group, miR-3612 inhibitor-NC group, miR-3612 inhibitor group, si-NC group, si-SEMA4C group, mimics-NC+pcDNA-NC group, miR-3612 mimics+pcDNA-NC group, and miR-3612 mimics+pcDNA- SEMA4C group. The corresponding nucleic acids and plasmids were transfected into each group of cells with transfection reagents.qPCR assay was used to detect the mRNA expression of miR-3612 and SEMA4C in hepatocellular carcinoma tissues and Hep3B and Huh7 cells. Dual-luciferase reporter gene assay and RNA immunoprecipitation assay (RIP) were used to validate the binding and regulatory relationship between miR-3612 and SEMA4C. qPCR and WB assays were used to detect the mRNA and protein expression of miR-3612 and SEMA4C in Hep3B and Huh7 cells after transfection in each group. The proliferation, migration and invasion abilities of Hep3B and Huh7 cells were detected by CCK-8 assay, cell scratch assay and Transwell assay, respectively. Results: miR-3612 was lowly expressed in hepatocellular carcinoma tissues and Hep3B and Huh7 cells (P<0.001), whereas SEMA4C was highly expressed (P<0.001). Overexpression of miR-3612 suppressed proliferation, migration, invasion, and expression of vimentin and SEMA4C proteins in Hep3B and Huh7 cells, promoted E-cadherin protein expression (P<0.05 or P<0.01 or P<0.001), and knockdown of miR-3612 promoted proliferation, migration, invasion and SEMA4C protein expression in Hep3B and Huh7 cells (P<0.05 or P<0.01 or P<0.001). Dual luciferase reporter gene assay and RIP assay confirmed that miR-3612 bound directly to SEMA4C (P<0.001), as indirectly evidenced by the negative correlation between miR-3612 and SEMA4C expression (P<0.001). Knockdown of SEMA4C significantly inhibited the proliferation, invasion and migration of Hep3B and Huh7 cells (P<0.05 or P<0.01 or P<0.001), and overexpression of SEMA4C reversed the inhibitory effect of miR-3612 overexpression on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of Hep3B and Huh7 cells (P<0.05 or P<0.01 or P< 0.001). Conclusion: miR-3612 affects the malignant biological behaviors of Hep3B and Huh7 cells by regulating SEMA4C expression, and it is expected to be a potential target for clinical hepatocellular carcinoma therapy.

国家自然科学基金（No. 81772180）；安徽省自然科学基金（No. KJ2016A722）