南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (1): 85-91.doi: 10.12122/j.issn.1673-4254.2023.01.11

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过表达CLEC5A基因抑制肝细胞癌增殖和转移并逆转上皮-间质转化

林 杰，区活辉，王卫东，马 靖，张伟杰，刘清波   

  1. 南方医科大学顺德医院（佛山市顺德区第一人民医院）肝胆胰脾外科，广东 佛山 528300
  • 出版日期:2023-01-20 发布日期:2023-02-22

Overexpression of CLEC5A inhibits cell proliferation and metastasis and reverses epithelial-mesenchymal transition in hepatocellular carcinoma

LIN Jie, OU Huohui, WANG Weidong, MA Jing, ZHANG Weijie, LIU Qingbo   

  1. Department of Hepatobiliary Surgery, Shunde Hospital, Southern Medical University (Shunde First People's Hospital), Foshan 528308, China
  • Online:2023-01-20 Published:2023-02-22

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摘要： 目的 研究C型凝集素结构域家族5成员A（CLEC5A）对肝细胞癌（HCC）增殖、迁移和侵袭能力以及上皮-间质转化（EMT）特性的影响，探索CLEC5A在HCC发生发展过程中的作用及机制。方法 采用免疫组化实验检测CLEC5A在50例HCC组织和癌旁组织中的表达特点，并分析其与HCC肝癌临床病理参数的相关性；通过慢病毒转染构建CLEC5A过表达HCC细胞，分别采用细胞增殖实验（CCK-8法、EdU法）、细胞迁移和侵袭实验（Transwell法）和Western blot实验检测CLEC5A对HCC细胞增殖、迁移及侵袭能力以及EMT标志物表达水平的影响。结果 CLEC5A蛋白在HCC癌组织中的表达低于癌旁组织，其表达水平与患者的肿瘤大小（P=0.008）、肿瘤数量（P=0.010）、肿瘤分化程度（P=0.016）、BCLC分期（P=0.040）和微血管侵犯（P=0.024）等相关；过表达CLEC5A能够抑制HCC细胞的增殖、迁移及侵袭能力，并逆转HCC细胞的EMT特性。结论 CLEC5A是HCC潜在的抑癌基因，有望成为该疾病新的治疗靶点。

关键词: 肝细胞癌；细胞增殖；迁移侵袭；上皮-间质转化

Abstract: Objective To evaluate the effects of CLEC5A expression level on cell proliferation, migration and invasion and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma (HCC) and explore the role of CLEC5A in the tumorigenesis and progression of HCC. Methods The expression level of CLEC5A was detected in 50 pairs of HCC and adjacent tissues using immunohistochemical staining, and its association with clinicopathological parameters of HCC patients was analyzed. Cultured HCC cell line SK-HEP-1 was transfected with a lentiviral vector overexpressing CLEC5A, and the transfection efficiency was verified using real-time fluorescence quantitative PCR and Western blotting. The changes in proliferation, migration and invasion abilities of the transfected cells were analyzed using CCK-8, 5-ethynyl-29-deoxyuridine (EdU) and Transwell assays, and EMT of the cells was determined using Western blotting. Results The protein expression level of CLEC5A was significantly lower in HCC tissues than in the adjacent tissues (P<0.001). The expression level of CLEC5A was significantly correlated with tumor size (P=0.008), tumor number (P=0.010), histological differentiation (P=0.016), microvascular invasion (P=0.024) and BCLC stage (P=0.040). In SK-HEP-1 cells, overexpression of CLEC5A obviously inhibited the cell proliferation, migration and invasion and reversed EMT phenotype of the cells. Conclusion CLEC5A is a potential HCC suppressor gene and may serve as a promising therapeutic target for HCC.

Key words: hepatocellular carcinoma; cell proliferation; cell invasion; epithelial-mesenchymal transition