Abstract：
Parecoxib, a well-recognized nonsteroidal anti-inflammatory drug, has been reported to possess anticancer properties in various tumor types. In this work, we aimed to investigate the potential anticancer effects of parecoxib on hepatocellular carcinoma (HCC) cells. To assess the impact of parecoxib on HCC cell proliferation, we employed Cell Counting Kit-8, colony formation, and 5-ethynyl-2\'-deoxyuridine assays. Hoechst/propidium iodide (PI) double staining and flow cytometry were performed to evaluate apoptosis and cell cycle analysis. Wound healing and transwell assays were utilized to assess cell migration and invasion. Tube formation assay was employed to analyze angiogenesis. Protein levels were determined using western blotting, and mRNA expression levels were assessed using quantitative real-time polymerase chain reaction (PCR). A xenograft mouse model was used to confirm the antitumor effects of parecoxib on HCC tumors in vivo. Our data demonstrated that parecoxib effectively inhibited the proliferation of HCC cells in a dose- and time-dependent manner. In addition, parecoxib induced cell cycle arrest in the G2 phase and promoted apoptosis. Moreover, parecoxib hindered tumor migration and invasion by impeding the epithelial-mesenchymal transition process. Further investigation showed that parecoxib could significantly suppress angiogenesis through the inhibition of extracellular signal-regulated kinase (ERK)-vascular endothelial growth factor (VEGF) axis. Notably, treatment with the ERK activator phorbol myristate acetate upregulated the expression of matrix metalloproteinase (MMP)-2, MMP-9, and VEGF and reversed the function of parecoxib in HCC cells. Besides, parecoxib displayed its antitumor efficacy in vivo. Collectively, our results suggest that parecoxib ameliorates HCC progression by regulating proliferation, cell cycle, apoptosis, migration, invasion, and angiogenesis through the ERK-VEGF/MMPs signaling pathway.
摘要：
帕瑞考昔布,一种公认的非甾体抗炎药，据报道，在各种肿瘤类型中具有抗癌特性。在这项工作中,我们旨在研究帕瑞昔布对肝细胞癌（HCC）细胞的潜在抗癌作用。为了评估帕瑞昔布对肝癌细胞增殖的影响，我们使用了细胞计数试剂盒-8，集落形成，和5-乙炔基-2'-脱氧尿苷测定。进行Hoechst/碘化丙啶（PI）双重染色和流式细胞术以评估细胞凋亡和细胞周期分析。利用伤口愈合和transwell测定来评估细胞迁移和侵袭。采用管形成测定来分析血管生成。蛋白质水平用蛋白质印迹法测定，使用定量实时聚合酶链反应（PCR）评估mRNA表达水平。使用异种移植小鼠模型来证实帕瑞昔布对体内HCC肿瘤的抗肿瘤作用。我们的数据表明，帕瑞昔布以剂量和时间依赖性方式有效抑制HCC细胞的增殖。此外,帕瑞昔布诱导细胞周期阻滞在G2期并促进细胞凋亡。此外，帕瑞昔布通过阻碍上皮-间质转化过程来阻碍肿瘤的迁移和侵袭。进一步研究表明，帕瑞昔布可以通过抑制细胞外信号调节激酶（ERK）-血管内皮生长因子（VEGF）轴，显着抑制血管生成。值得注意的是,用ERK激活剂佛波醇肉豆蔻酸盐醋酸盐处理可上调HCC细胞中基质金属蛋白酶（MMP）-2，MMP-9和VEGF的表达，并逆转帕瑞昔布的功能。此外,帕瑞昔布在体内显示出其抗肿瘤功效。总的来说，我们的结果表明，帕瑞昔布通过调节增殖来改善HCC的进展，细胞周期,凋亡，迁移,入侵,和血管生成通过ERK-VEGF/MMPs信号通路。
