Department of General Medicine, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People's Republic of China.
Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, People's Republic of China.
Drug Des Devel Ther. 2023 Mar 30;17:993-1006. doi: 10.2147/DDDT.S402824. eCollection 2023.
To evaluate the effect of 5-fluorouracil (5-FU) combined with rutaecarpine (RUT) on the antiproliferative, anti-migratory, and apoptosis-promoting ability of colorectal cancer (CRC) cells and explore the underlying mechanism.
The antiproliferative effects of RUT and 5-FU on CRC cells were evaluated using MTT and colony formation assays. Anti-migration was assessed by cell scratch and transwell tests. The synergistic effect of RUT and 5-FU was assessed by isobologram and combination index analysis using CompuSyn software. The effects of RUT and 5-FU on cell apoptosis were detected by flow cytometry. Differences in protein expression levels with or without RUT and/or 5-FU treatment were assessed by Western blot. Moreover, a mouse xenograft model of CRC was established to investigate the antitumor effect of RUT and 5-FU in vivo, and Ki67 and cleaved caspase-3 expression was detected by immunofluorescence.
In this study, we found that 5-FU combined with RUT can inhibit the proliferative, migratory, and antiapoptotic abilities of CRC cells to a significantly greater extent than either RUT or 5-FU alone both in vivo and in vitro. Western blot analysis showed that the level of signal transducer and activator of transcription 3 (STAT3) phosphorylation in CRC cells was significantly reduced after combination therapy compared with that seen with the respective monotherapies. In addition, combination therapy influenced the STAT3 signaling pathway, namely, it inhibited the expression of c-Myc, CDK4, and Bcl-2 while enhancing that of the proapoptotic protein cleaved caspase-3. Immunofluorescence staining further showed that the expression of Ki67 and cleaved caspase-3 was significantly downregulated and upregulated, respectively, in tumor tissues of mice treated with combination therapy compared with that observed with 5-FU treatment alone.
Combined therapy with 5-FU and RUT exerted a superior curative effect in CRC than treatment with either single drug alone and has potential as a novel therapeutic modality for the treatment of CRC.
评估 5-氟尿嘧啶（5-FU）联合吴茱萸碱（RUT）对结直肠癌细胞增殖、迁移和促凋亡能力的影响，并探讨其潜在机制。
采用 MTT 和集落形成实验评估 RUT 和 5-FU 对 CRC 细胞的增殖抑制作用。通过细胞划痕和 Transwell 实验评估抗迁移作用。使用 CompuSyn 软件的等效应图和组合指数分析评估 RUT 和 5-FU 的协同作用。通过流式细胞术检测 RUT 和 5-FU 对细胞凋亡的影响。通过 Western blot 检测有无 RUT 和/或 5-FU 处理时蛋白表达水平的差异。此外，建立 CRC 小鼠异种移植模型，研究 RUT 和 5-FU 的体内抗肿瘤作用，并通过免疫荧光检测 Ki67 和 cleaved caspase-3 的表达。
本研究发现，5-FU 联合 RUT 可显著抑制 CRC 细胞的增殖、迁移和抗凋亡能力，无论是在体内还是体外，其效果均明显强于单独使用 RUT 或 5-FU。Western blot 分析显示，与单独用药相比，联合治疗后 CRC 细胞中信号转导和转录激活因子 3（STAT3）磷酸化水平显著降低。此外，联合治疗还影响了 STAT3 信号通路，即抑制了 c-Myc、CDK4 和 Bcl-2 的表达，同时增强了促凋亡蛋白 cleaved caspase-3 的表达。免疫荧光染色进一步显示，与单独使用 5-FU 治疗相比，联合治疗组小鼠肿瘤组织中 Ki67 和 cleaved caspase-3 的表达分别显著下调和上调。
与单独使用单一药物相比，5-FU 联合 RUT 治疗结直肠癌具有更好的疗效，有望成为治疗结直肠癌的一种新的治疗方法。
