阿帕替尼与5-氟尿嘧啶联合应用对人胃癌细胞AGS增殖、凋亡及迁移的影响

目的:探索小分子酪氨酸激酶抑制剂阿帕替尼与5-氟尿嘧啶(5-fluorouracil,5-FU)联合应用对人胃癌细胞AGS增殖、凋亡及迁移的影响。方法:采用Western Blot方法评估人胃癌细胞系AGS、MKN-45、SGC-7901及人脐静脉血管内皮细胞中血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)蛋白表达情况。采用MTS法测定空白对照组、阿帕替尼单药组、5-FU单药组及阿帕替尼与5-FU联合组对人胃癌细胞AGS增殖情况的影响;采用Annexin V-FITC/PI流式双染法测定四组细胞的凋亡情况;采用划痕试验测定药物处理后人胃癌细胞AGS的迁移情况。结果:人胃癌细胞AGS表达VEGFR2。MTS结果显示,与空白对照、阿帕替尼单药和5-FU单药比较,阿帕替尼与5-FU联合应用可显著抑制人胃癌细胞AGS增殖,差异均有统计学意义(P<0.05);阿帕替尼单药、5-FU单药对人胃癌细胞AGS增殖的抑制呈时间及剂量依赖性。流氏双染法结果显示,与空白对照、阿帕替尼单药和5-FU单药比较,阿帕替尼与5-FU联合应用可显著诱导人胃癌细胞AGS凋亡,差异均有统计学意义(P<0.001);其总凋亡率为45.8%,以晚期调亡细胞为主。划痕试验结果显示,与空白对照、阿帕替尼单药和5-FU单药比较,阿帕替尼与5-FU联合应用可明显抑制人胃癌细胞AGS迁移,差异均有统计学意义(P<0.001)。结论:体外实验结果显示,阿帕替尼与5-FU联合应用可显著抑制人胃癌细胞AGS的增殖、迁移,并诱导细胞凋亡,该研究可为上述两药联合应用治疗晚期胃癌提供实验依据。

Effect of Apatinib Combined with 5-Fluorouracil on the Proliferation,Apoptosis and Migration of Gastric Cancer AGS Cells

OBJECTIVE:To probe into the effect of apatinib combined with 5-fluorouracil(5-FU)on the proliferation,apoptosis and migration of gastric cancer AGS cells.METHODS:Western Blot method was adopted to evaluate the expression levels of vascular endothelial growth factor receptor 2(VEGFR2)in human gastric cancer AGS,MKN-45,SGC-7901 cells and human umbilical vein endothelial cell(HUVEC).MTS method was used to determine the effect of blank control group,apatinib group,5-FU group and apatinib combined with 5-FU group on the proliferation of human gastric cancer AGS cells;Annexin V-FITC/PI staining flow cytometry was used to determine the apoptosis of the four groups of cells;Scratch test was used to determine the migration of AGS cells after drug treatment.RESULTS:Human AGS cells expressed VEGFR2.MTS results showed that compared with blank control group,apatinib group and 5-FU group,the combination of apatinib and 5-FU can significantly inhibit the proliferation of human gastric cancer AGS cells,with statistically significant differences(P<0.05);the inhibition of apatinib alone and 5-FU alone on the proliferation of AGS cells was in a time-dependent and dose-dependent.The results of double staining method showed that compared with blank control group,apatinib group and 5-FU group,the combination of apatinib and 5-FU can significantly induce the apoptosis of human gastric cancer AGS cells,with statistically significant differences(P<0.001);the total apoptosis rate was 45.8%,mainly were advanced apoptotic cells.The results of scratch test showed that compared with blank control group,apatinib group and 5-FU group,the combination of apatinib and 5-FU can significantly inhibit the migration of AGS cells,with statistically significant differences(P<0.001).CONCLUSIONS:Results of experiments in vitro indicate that the combination of apatinib and 5-FU can significantly inhibit the proliferation and migration of human gastric cancer AGS cells,and induce cell apoptosis.This study can provide experimental evidence for the combined application of the above two drugs in the treatment of advanced gastric cancer.