小白菊内酯对胃癌细胞BGC-823的抑制作用及其机制

INHIBITORY EFFECT OF PARTHENOLIDE ON GASTRIC CANCER BGC-823 CELLS AND RELATED MECHANISM

目的研究小白菊内酯(PN)对胃癌细胞BGC-823的抑制作用及其机制。方法将BGC-823细胞分为DMSO组和PN组,DMSO组常规进行细胞培养,PN组则采用不同浓度的PN处理细胞。两组细胞采用CCK-8技术检测不同时间点细胞活力的变化,采用克隆形成实验检测细胞增殖能力的变化,采用流式细胞术检测细胞不同周期比例及凋亡水平,采用荧光实验检测细胞内活性氧簇(ROS)生成水平,采用Western Blot方法检测细胞周期及凋亡相关蛋白水平的变化。结果与DMSO组比较,随着PN浓度的增加BGC-823细胞的活力及增殖能力均下降(F=283.80、312.60,P<0.01)。24h时,PN组的总凋亡比例与DMSO组相比明显升高(t=27.57,P<0.01),且随着处理时间的延长凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶(caspase)8、caspase9和caspase3剪切型的表达明显增加(F=228.10～2 024.00,P<0.01)。与DMSO组相比较,PN组细胞周期被阻滞在G1期(t=4.22、7.44,P<0.05);而且随着处理时间的延长,细胞周期蛋白(Cyclin)D1及CyclinE1水平明显下降(F=159.40、25.40,P<0.01),P53及P21蛋白表达水平明显升高(F=53.74、73.03,P<0.01)。24h时,PN组细胞内ROS生成水平较DMSO组明显升高(t=10.92,P<0.01);而且随着处理时间的延长,细胞核内c-Myc基因、转录因子E2F1、核因子κB(NF-κB)以及磷酸化的信号传导及转录激活因子3(p-STAT3)蛋白的表达水平明显下降(F=306.90、1 067.00,t=8.72、9.15,P<0.01)。结论 PN对BGC-823细胞增殖能力呈剂量依赖性抑制,对细胞周期产生阻滞并诱导细胞凋亡,这种抗肿瘤作用可能是通过抑制STAT3-c-Myc-E2F1发挥作用的。

Objective To investigate the inhibitory effect of parthenolide(PN)on gastric cancer BGC-823 cells and related and mechanism. Methods BGC-823 cells were divided into DMSO group and PN group.The cells in the DMSO group were cultured with the conventional method,and those in the PN group were treated with different concentrations of PN.CCK-8 assay was used to measure the change in cell viability at different time points,colony-forming assay was used to measure the change in the proliferative capacity of BGC-823 cells,flow cytometry was used to measure the proportion of cells in different cell cycles and the apoptosis of cells,fluorescence assay was used to measure the level of reactive oxygen species(ROS)in cells,and Western Blot was used to measure the changes in the levels of cell cycle-and apoptosis-related proteins. Results Compared with the DMSO group,the PN group had significant reductions in the viability and proliferative capacity of BGC-823 cells in a concentration-dependent manner(F=283.80,312.60,P<0.01).At 24 hof PN exposure,compared with the DMSO group,the PN group had a significant increase in the proportion of apoptotic cells(t=27.57,P<0.01),as well as significant increases in the expression of the apoptosis-related proteins cleaved caspase-8,cleaved caspase-9,and cleaved caspase-3 over the time of treatment(F =228.10-2 024.00,P<0.01).Compared with the DMSO group,the PN group had a significantly higher number of cells arrested in G1 phase(t=4.22,7.44,P<0.05),as well as significant reductions in the levels of CyclinD1 and CyclinE1(F=159.40,25.40,P<0.01)and significant increases in the protein expression of P53 and P21(F=53.74,73.03,P<0.01)over the time of treatment.Compared with the DMSO group at 24 hof PN exposure,the PN group had a significant increase in the level of ROS(t=10.92,P<0.01),and over the time of treatment,the PN group had significant reductions in the protein expression of c-Myc,the transcription factor E2 F1,nuclear factor-kappa B,and phosphorylated STAT3(F=306.90,1 067.00,t=8.72,9.15,P<0.01). Conclusion PN can inhibit the proliferation of BGC-823 cells,induce G1-phase cell cycle arrest,and thus exerts an antitumor effect in a dose-dependent manner,possibly by inhibiting the STAT3-c-Myc-E2 F1 axis.