目的：分析环氧合酶2（cyclooxygenase-2，COX-2）的表达与肝癌患者临床病理特征及预后的关系，探讨刺芒柄花素 （formononetin，FOR）在肝癌发生和发展中的作用及其机制。方法：收集2021年1~5月河北医科大学第四医院20例手术治疗肝 癌患者的癌和相应的癌旁组织、60例手术治疗肝癌患者的临床资料，以及人肝癌细胞系HepG2和Bel-7402。用qPCR法和免疫组 织化学染色法检测肝癌组织中COX-2的表达，分析其表达与患者临床病理特征和预后的关系。构建小鼠二乙基亚硝胺诱导 肝癌模型，验证FOR对肝癌发病的影响。用0.003、0.006 μmol/ml的FOR分别处理肝癌细胞HepG2和Bel-7402 48 h后，用CCK-8 法和流式细胞术检测FOR对肝癌细胞增殖和周期的影响，qPCR和WB法检测细胞中COX-2、CDK4、CDK6和cyclin D1表达的变 化。结果：肝癌组织中COX-2 mRNA表达水平显著高于癌旁组织（P<0.01），COX-2 蛋白表达阳性率为68.3%（41/60）；COX-2 表达阳性与患者 TNM 分期晚、肿瘤大、生存期短有关（均P<0.05）。诱导小鼠肝癌模型中，FOR处理组小鼠肝癌的发生率显著 降低、肝癌组织中COX-2表达显著降低（均P<0.05）。FOR处理可显著抑制HepG2和Bel-7402细胞的增殖能力，增加G0/G1期细 胞比例、降低S和G2期细胞比例（均P<0.05），抑制细胞中COX-2和cyclin D1的表达（均P<0.01）。。结论：COX-2表达阳性肝癌患 者临床分期较晚且预后较差，FOR可通过抑制COX-2和cyclin D1表达来降低肝癌的发生和发展。

Objective: To analyze the relationship between the expression of cyclooxygenase-2 (COX-2) and clinicopathological features and prognosis of patients with liver cancer, and to explore the role and mechanism of formononetin (FOR) in the pathogenesis of liver cancer. Methods: The clinical data of 60 patients with liver cancer and 20 pairs of surgically resected cancer tissues and corresponding para-cancerous tissues from liver cancer patients that treated in the Fourth Hospital of Hebei Medical University during January 2021 and May 2021 were collected for this study; in addition, human liver cancer cell lines HepG2 and Bel-7402 were also collected. The expression of COX-2 in liver cancer tissues was detected by qPCR and immunohistochemical staining, and the relationship between COX-2 expression and clinicopathological characteristics and prognosis of patients was analyzed. Diethylnitrosamine-induced mouse model of liver cancer was established to verify the effect of FOR on the incidence of liver cancer. After being treated with 0.003 and 0.006 μmol/ml FOR 48 h, the effects of FOR on proliferation and cell cycle of HepG2 and Bel-7402 cells were detected by CCK-8 and Flow cytometry. The changes in the expression of COX-2, CDK4, CDK6 and cyclin D1 were detected by qPCR and WB. Results: The mRNA expression of COX-2 in liver cancer tissues was significantly higher than that in para cancerous tissues ( P<0.01 ). The positive expression rate of COX-2 in liver cancer tissues was 68.3% (41/60), and the positive expression of COX-2 was correlated with advanced TNM stage, large tumor and short survival (all P<0.05 ). In the xenograft mouse model of induced liver cancer, the incidence of liver cancer in mice treated with FOR was significantly reduced, and the expression of COX-2 in liver tissues was significantly decreased (all P<0.05 ). FOR significantly inhibited the proliferation, increased the proportion of cells at G0/G1 phase, decreased the proportion of cells at S and G2 phase (all P<0.05 ), and inhibited the expression of COX-2 and cyclin D1 in HepG2 and Bel-7402 cells (all P<0.01 ). Conclusion: Liver cancer patients with positive COX-2 expression have an advanced clinical stage and a poor prognosis. FOR can prevent the occurrence and development of liver cancer by inhibiting the expression of COX-2 and cyclin D1.

河北省卫生厅科研基金资助项目（No. 20200110）