芹菜素对人结肠癌细胞SW620增殖与葡萄糖摄取的影响

Effects of Apigenin on Proliferation of Human Colon Cancer Cells and Glucose Uptake

目的探讨芹菜素对人结肠癌细胞SW620细胞增殖与葡萄糖摄取的影响。方法以SW620细胞为研究对象,用10~80μmol·L^(-1)不同浓度芹菜素作用于细胞,2.5 mmol·L^(-1)的二氟甲基鸟氨酸(DFMO)作为阳性对照药,细胞以5×103个·mL^(-1)的密度接种于96孔培养板,每组设6个复孔,分别培养24,48,72 h后,CCK-8法检测细胞增殖抑制情况;同时,取96孔板培养24,48,72 h的细胞上清液,用葡萄糖测定试剂盒检测SW620细胞葡萄糖摄取情况;细胞以2×106个·mL^(-1)的密度接种于60 mm的中皿中,培养48 h后,Western blot及荧光定量PCR检测葡萄糖转运蛋白1(GLUT1)及钠-葡萄糖共转运载体(SGLT1)及其mRNA表达水平的变化。结果与对照组比较,芹菜素各浓度组均能抑制SW620细胞的增殖,且抑制作用呈时间和浓度依赖性,差异有统计学意义(P<0.05,P<0.01);芹菜素各浓度组作用后SW620细胞对葡萄糖的摄取明显减少(P<0.05,P<0.01);与对照组比较,GLUT1和SGLT1蛋白及m RNA随着芹菜素浓度的升高表达减少(P<0.05,P<0.01)。结论芹菜素具有抑制人结肠癌细胞SW620细胞增殖与葡萄糖摄取的作用,这为结肠癌的治疗提供了新的思路和方法。

Objective To investigate the effect of apigenin on the proliferation of human colon cancer cells SW620 and glucose uptake.Methods SW620 cells were treated with apigenin at 10~80 μmol·L^-1and 2.5 mmol·L^-1DFMO was used as the positive control drug.The cells were inculated at the density of 5×103·mL^-1in 96-well plate,6 duplicate wells for each group.After culturing for 24,48,72 h,cell proliferation was determinated by CCK8.Meanwhile,the supernatant of the SW620 cells cultured in 96-well plate for 24,48,72 h was used for the detection of glucose uptake with glucose test kit.In the other experiment,the cells were inoculated at the density of 2×10~6·mL^-1in 60-mm culture dishes for48 h,and then the protein and mRNA levels of glucose transporter 1（GLUT1）and sodium/glucose cotransporter^-1（SGLT1） were detected by Western blotting and qPCR respectively.Results Compared with the control group,apigenin at various concentrations significantly inhibit the proliferation of human colon cancer cells SW620 in a time-and concentration-dependent manner,and the difference was statistically significant（P〈0.05,P〈0.01）.Apigenin at different concentrations also significantly reduced the intake of glucose by SW620 cells（P〈0.05,P〈0.01）.Compared with the control group,the expression levels of GLUT1 and SGLT1 protein and m RNA were decreased with the increase of apigenin concentration（P〈0.05,P〈0.01）.Conclusion Apigenin has an effect on inhibiting the proliferation and reducing the glucose uptake of human colon cancer cells SW620,which will provide a new approach and method for the treatment of colon cancer.