二苯乙烯苷调控MAPKs信号通路诱导大肠癌SW116细胞凋亡

2,3,5,4′-etrahydroxy stilbene-2-O-β-D-glucoside (THSG) induces apoptosis of SW116 cells via regulating MAPKs signaling pathway in colorectal cancer

目的研究二苯乙烯苷(THSG)对大肠癌SW116细胞凋亡的诱导作用及其可能的分子机制。方法体外常规培养SW116细胞,采用不同剂量(0、5、10、20、40、80、120mmol/L)THSG处理SW116细胞,CCK8法分析不同剂量药物处理24、48、72h后细胞的存活率;不同浓度(0、10、20、40mmol/L)THSG干预SW116细胞24h后,Annexin/PI双染后流式细胞仪检测SW116细胞凋亡情况,Western blot方法检测SW116细胞中凋亡相关蛋白PARP和Pro-caspase 3的表达及MAPKs信号通路p38、ERK、JNK的磷酸化水平;p38、ERK、JNK特异性抑制剂干预细胞后,Western blot方法检测抑制剂对p38、ERK、JNK磷酸化的抑制作用。结果THSG呈剂量和时间依赖性地抑制大肠癌SW116细胞的增殖,诱导细胞凋亡;THSG干预SW116细胞后,Cleaved PARP表达显著升高、Pro-caspase 3表达显著降低,胞内p38和JNK磷酸化水平显著升高(P<0.05),然而ERK磷酸化水平显著降低(P<0.05)。结论特异性抑制剂阻断ERK激活,可以增强THSG诱导SW116细胞凋亡,特异性阻断p38、JNK活化,能够部分逆转THSG对p38和JNK磷酸化和细胞凋亡的诱导作用。THSG通过激活p38和JNK信号传导,抑制ERK信号途径,诱导大肠癌SW116细胞凋亡。

Objective To study the effect of 2,3 ,5 ,4 '-etrahydroxy stilbene-2-O-β-D-glucoside ( THSG) on apoptosis of colorectal cancer cells ( SW116) and explore the molecular mechanism. Methods SW116 cells were cultured in vitro. Cells were treated with varied doses of THSG ( 0,5,10,20,40,80 and 120 mmol /L) for 24,48 and 72 h, and the cell survival was detected by CCK8 assay. After SW116 cells were treated with varied doses of THSG ( 0, 10, 20 and 40 mmol /L) for 24 h, cell apoptosis was determined by Annexin /PI methods. Western blot was used to detect the expression levels of PARP,Pro-caspase 3 and the phosphorylation of p38,ERK and JNK. The cells were treated with specific inhibitors of p38,ERK and JNK, and the inhibitory effects on these pathways were detected with Western blot. Results THSG inhibited the proliferation and induced apoptosis of SW116 cells in dose-and time-dependent methods. THSG treatment obvious enhanced the expression of Cleaved PARP and phosphorylation of p38 and JNK,but decreased the expression of Pro-caspase 3 and ERK phosphorylation in the cells ( P < 0. 05). Conclusion Blocking ERK activation with the ERK inhibitor obviously enhances THSG-induced SW116 cell apoptosis,while inhibiting p38 and JNK activation partly reverses THSG-induced p38 and JNK phosphorylation and apoptosis in the cells. THSG induces apoptosis of colorectal cancer cells ( SW116) in vitro by activating p38 and JNK signaling pathways and inhibiting ERK signaling pathway.