小白菊内酯联合5-氟尿嘧啶用药对结直肠癌细胞自噬的影响

Effect of Parthenolide Combined with 5-fluorouracil on Autophagy in Colorectal Cancer Cells

目的探讨小白菊内酯(PT)与5-氟尿嘧啶(5-FU)联合用药对结直肠癌(CRC)细胞活性和自噬的影响。方法采用CCK-8法检测不同浓度PT对结直肠癌细胞HCT 116活性的影响,并检测PT联合5-FU对HCT 116细胞活性的影响。采用蛋白质印迹法检测5-FU处理以及PT联合5-FU对HCT 116后CRC细胞自噬相关蛋白LC3Ⅱ/Ⅰ水平的变化;免疫荧光检测LC3斑点数目变化情况。运用自噬抑制3-MA处理PT与5-FU的联合运用,CCK-8法检测细胞活性,蛋白质印迹法检测LC3Ⅱ/Ⅰ自噬表达情况,免疫荧光检测LC3斑点数目变化情况。结果与5-FU单独处理比较,低浓度PT与5-FU联合运用后显著抑制CRC细胞的活性(P<0.05);蛋白质印迹法检测PT促进5-FU诱导的自噬,免疫荧光可见PT与5-FU联合运用后LC3荧光斑点明显增多;运用自噬阻断剂3-MA后抑制了自噬,降低了PT与5-FU联合运用对CRC细胞活性的抑制作用。结论PT促进5-FU诱导的细胞自噬,并抑制CRC细胞的活性,而抑制自噬可以降低PT和5-FU联合对CRC细胞活性的抑制作用。

Objective To investigate the effect of Parthenolide(PT)combined with 5-fluorouracil(5-FU)on the activity and autophagy of colorectal cancer cells.Methods The CCK-8 method was used to detect the effect of different concentrations of PT on the activity of colorectal cancer cells HCT 116,and to detect the effect of PT combined with 5-FU on the activity of HCT 116 cells.Western blotting was used to detect the changes in the level of autophagy-related protein LC3Ⅱ/Ⅰin CRC cells after 5-FU treatment and PT combined with 5-FU on HCT 116;the changes in the number of LC3 spots were detected by immunofluorescence.Autophagy inhibition 3-MA was used to treat the combined application of PT and 5-FU,CCK-8 method was used to detect cell viability.Western blot method was used to detect the autophagy expression of LC3Ⅱ/Ⅰ,and immunofluorescence was used to detect the number of LC3 spots.Results Compared with 5-FU treatment alone,the combination of low-concentration PT and 5-FU significantly inhibited the activity of CRC cells(P<0.05);Western blot detection of PT promoted 5-FU-induced autophagy,immunofluorescence showed that after the combined use of PT and 5-FU,the LC3 fluorescent spots increased significantly;the use of the autophagy blocker 3-MA inhibited autophagy and reduced the inhibitory effect of the combined use of PT and 5-FU on CRC cell activity.Conclusion PT can promote 5-FU-induced autophagy and inhibit the activity of CRC cells,while inhibition of autophagy can reduce the function of PT and 5-FU on the activity of CRC cells.