姜黄素增强氟尿嘧啶诱导胃癌SGC-7901细胞凋亡的效应与Fas信号通路的研究

Study on the enhancing effect of curcumin on fluorouracil induced apoptosis of gastric cancer SGC-7901 cells and related Fas signaling pathway

目的探索姜黄素(CUR)对氟尿嘧啶(5-FU)诱导胃癌SGC-7901细胞凋亡的增强效应及其作用机理。方法将体外培养的胃癌SGC-7901细胞分为6组,即空白对照组、CUR组、5-FU低剂量+CUR组、5-FU中剂量组、5-FU中剂量+CUR组、5-FU高剂量组。应用AO/EB双染荧光显微镜观察法检测各组细胞发生凋亡的形态学改变;应用Annexin V-FITC/PI双染的流式细胞术检测各组细胞的早期与晚期凋亡率;最后采用Western blot方法检测各组细胞凋亡相关的Fas通路中Fas、FADD、caspase-8、caspase-3蛋白的表达水平。结果荧光显微镜检测显示,与空白对照组比较,各药物处理组SGC-7901细胞发生凋亡的数量显著增多(P<0.05),其中5-FU低剂量+CUR组的凋亡率显著高于5-FU中剂量组(P<0.05),5-FU中剂量+CUR组的凋亡率极显著高于5-FU高剂量组(P<0.01);流式细胞检测结果显示,与空白对照组比较,各药物处理组的细胞凋亡率均显著升高(P<0.05);Western blot检测显示,与空白对照组比较,药物处理24h的细胞,凋亡通路中的Fas、FADD、caspase-8、caspase-3的表达水平均显著增加(P<0.05)。结论 CUR对5-FU诱导胃癌SGC-7901细胞凋亡的作用具有增强效应,其作用机理可能是通过上调Fas、FADD、caspase-8以及caspase-3蛋白的表达而实现的。

Objective To explore the enhancing effect of curcumin(CUR)on fluorouracil(5-FU)induced apoptosis of gastric cancer SGC-7901 cells and its mechanism. Methods Apoptosis of SGC-7901 cells induced by curcumin combined with fluorouracil was observed under fluorescence microscope based onAO/EB staining. The effect of curcumin combined with fluorouracil on the apoptosis of gastric cancer SGC-7901 cells was detected by flow cytometry based on Annexin V-FITC/PI double staining. The expressions of Fas,FADD,caspase-8 and caspase-3 in SGC-7901 cells apoptosis signalingpathway were detected by Western blot(WB). Results Compared with that of control group,the number of apoptosis in SGC-7901 cells was significantly increased(P<0.05)and showed dose-dependent and time-dependent effects;curcumin(30 M)combined with low dose(10 M)or medium dose(20 M)fluorouracil can achieve the effect of medium dose(20 M)and high dose(40 M)fluorouracil alone,respectively;the results of flow cytometry showed that the apoptosis rate in experimental group was significantlyhigher than that in control group(P<0.05)and showed dose and time-dependent effect;the results of WB method showed that,compared with control group,the expressions of Fas,FADD,caspase-8 and caspase-3,which are closely related to apoptosis,were significantly increased in experimental groups after being treated with CUR and fluorouracil at different doses for 24 h(P<0.05). Conclusion CUR can enhance the apoptosis of SGC-7901 cells induced by fluorouracil by upregulating the expressions of Fas,FADD,caspase-8 and caspase-3.