2023 年第 9 期 第 18 卷

氟尿嘧啶联合低剂量雷公藤内酯对胃癌细胞增殖和迁移及侵袭的影响及机制研究

Effects and mechanism of fluorouracil combined with low-dose triptolide on proliferation, migration and invasion of gastric cancer cells

作者：杜艳明崔丽伟檀靖宇马召雨马丽丽

英文作者：Du Yanming Cui Liwei Tan Jingyu Ma Zhaoyu Ma Lili

单位：河北省唐山市人民医院消化科，唐山063000

英文单位：Department of Gastroenterology Tangshan People′s Hospital Hebei Province Tangshan 063000 China

关键词：胃癌；氟尿嘧啶；雷公藤内酯；增殖；迁移；侵袭

英文关键词：Gastriccancer;Fluorouracil;Triptolide;Proliferation;Migration;Invasion

• 摘要：

• 目的  探讨氟尿嘧啶联合雷公藤内酯是否可协同抑制胃癌细胞增殖和迁移及侵袭，并进一步分析可能的作用机制。方法 将体外培养的胃癌SGC-7901细胞依据添加药物的不同分为对照组（不加药处理）、氟尿嘧啶组（加5 mg/L的氟尿嘧啶）、雷公藤内酯组（加2 μg/L的雷公藤内酯）、联合组（加5 mg/L的氟尿嘧啶和2 μg/L的雷公藤内酯）。观察噻唑蓝实验检测氟尿嘧啶（0、5、10、20和40 mg/L）、雷公藤内酯（0、2、4、8和16 μg/L）对胃癌SGC-7901细胞增殖的抑制作用，并计算细胞生存率和联合作用指数。采用细胞划痕实验、Transwell实验、凋亡实验和蛋白质印迹法分别检测氟尿嘧啶、雷公藤内酯单独或联合对SGC-7901细胞迁移率、细胞侵袭率、细胞凋亡率、磷酸酶2A的癌性抑制因子（CIP2A）、蛋白磷酸酶2A（PP2A）、磷酸化-哺乳动物雷帕霉素靶标复合物1(p-mTORC1)、哺乳动物雷帕霉素靶标复合物1（mTORC1）蛋白表达的影响。结果 氟尿嘧啶和雷公藤内酯均呈剂量依赖性抑制SGC-7901细胞增殖，半数抑制浓度分别为15.3 mg/L、10.3 μg/L，二者不同浓度联用联合作用指数值均小于1。联合组细胞迁移率、细胞侵袭率低于氟尿嘧啶组和雷公藤内酯组［（5.2±1.3）%比（33.6±2.7）%、（38.5±3.1）%，（19.3±2.5）%比（57.2±3.5）%、（80.4±2.8）%］，差异均有统计学意义（均P<0.05）。联合组细胞凋亡率高于氟尿嘧啶组和雷公藤内酯组（均P<0.05）。联合组细胞CIP2A、PP2A、p-mTORC1蛋白相对表达水平低于对照组、氟尿嘧啶组和雷公藤内酯组，差异均有统计学意义（均P<0.05）。结论 氟尿嘧啶联合雷公藤内酯可协同抑制SGC-7901细胞增殖、迁移和侵袭，可能与抑制CIP2A/PP2A/mTORC1信号通路传导有关。

• Objective To investigate whether fluorouracil combined with triptolide can synergistically inhibit the proliferation, migration and invasion of gastric cancer cells, and to analyze the possible mechanism. Methods In vitro cultured gastric cancer SGC-7901 cells were divided into control group(without drug treatment), fluorouracil group(with 5 mg/L fluorouracil), triptolide group(with 2 μg/L triptolide), combined group(with 5 mg/L fluorouracil and 2 μg/L triptolide). Methyl thiazolyl tetrazolium assay was observed to detect the inhibitory effects of fluorouracil(0, 5, 10, 20 and 40 mg/L) and triptolide(0, 2, 4, 8 and 16 μg/L) on the proliferation of gastric cancer SGC-7901 cells, and the cell survival rate and the combined effect index were calculated. Cell scratch assay, Transwell assay, apoptosis assay and western blot assay were used to detect the effects of fluorouraciland triptolide alone or in combination on SGC-7901 cell migration rate, cell invasion rate, cell apoptosis rate, and expressions of cancerous inhibitor of protein phosphatase 2A(CIP2A), protein phosphatase 2A (PP2A), phosphorylation-mechanistic target of rapamycin complex 1 (p-mTORC1), and mechanistic target of rapamycin complex 1 (mTORC1). Results Both fluorouracil and triptolide inhibited the proliferation of SGC-7901 cells in a dose-dependent manner, with median inhibitory concentration values of 15.3 mg/L and 10.3 μg/L, respectively, and combined effect index values when combined with different concentrations of the two were both less than 1. The cell migration rate and cell invasion rate in the combined group were lower than those in the fluorouracil group and the triptolide group［(5.2±1.3)% vs (33.6±2.7)%, (38.5±3.1)%; (19.3±2.5)% vs (57.2±3.5)%, (80.4±2.8)%］(all P<0.05). The cell apoptosis rate in the combined group was higher than those in the fluorouracil group and the triptolide group(both P<0.05). The relative expression levels of CIP2A, PP2A and p-mTORC1 protein in the combined group were lower than those in the control group, the fluorouracil group and the triptolide group(all P<0.05). Conclusions  Fluorouracil combined with triptolide can synergistically inhibit the proliferation, migration and invasion of SGC-7901 cells, which may be related to the inhibition of CIP2A/PP2A/mTORC1 signaling transduction.