青蒿琥酯对胃癌细胞系HGC27细胞增殖、凋亡的影响及其机制

Effects of artesunate on cell proliferation and apoptosis in human gastric cancer HGC27 cells and its mechanisms

目的 探讨青蒿琥酯对大胃癌细胞系HGC27细胞增殖、凋亡的影响及其机制.方法 体外培养HGC27细胞,采用不同浓度青蒿琥酯处理24、48、72 h,MTT法测定其对HGC27细胞增殖的影响;倒置显微镜下观察细胞的形态学变化;青蒿琥酯（浓度分别为20、40、80 mg/L）处理HGC27细胞48 h后,分别采用流式细胞仪检测细胞凋亡、分光光度计检测Casepase-3、Caspase-9相对活性、Western blot法检RUNX-3蛋白表达情况.结果 青蒿琥酯（浓度10～100 mg/L）能抑制HGC27细胞的增殖,呈剂量和时间依赖性;倒置显微镜下可观察到典型的细胞凋亡形态.青蒿琥酯（浓度分别为20、40、80 mg/L）作用HGC27细胞48 h后,细胞凋亡率分别为11.5％、21.4％、36.6％,而对照组凋亡率仅为2.2％;处理后Casepase-3相对活性分别为（0.19±0.02）、（0.25±0.04）和（0.31±0.03）,对照组为（0.11±0.02）,Casepase-9相对活性分别为（0.18±0.02）、（0.23±0.03）和（0.30±0.04）,对照组为（0.10±0.02）,与对照组比较,青蒿琥酯处理组Casepase-3、Caspase-9相对活性显著增加,差异均有统计学意义（均P＜0.05）;RUNX-3蛋白表达上调,呈剂量依赖性.结论 青蒿琥酯能抑制HGC27细胞的增殖并诱导其凋亡,其作用机制可能与增加细胞Casepase-3、Caspase-9活性、上调RUNX-3蛋白的表达有关.青蒿琥酯是一种前景广阔的抗肿瘤药物.

Objective To investigate the effect of artesunate on cell proliferation and apoptosis in gastric cancer line HGC27 cells and discuss its possible mechanisms. Methods HGC27 cells were cultured in vitro. After treatment by artesunate at different concentrations respectively at 24, 48, 72 h, the cell survival was determined by the MTY method. The changes of cell morphology were observed by inverted microscope. After 48 h treatment by artesunate （20, 40, 80 mg/L）, the HGC27 cell apoptosis was detected by flow eytometry, the relative activity of Caspase-3 and Caspase-9 was monitored by spectrophotometer, the change of protein expression of RUNX-3 was detected by western blot. Results From the data of MTT, the cell proliferation of human gastriel cancer HGC27 cells was inhibited by artesunate （10-100 rag/L） in a dose-dependent and time-dependent manner. Typical apoptosis morphology of HGC27 cells was observed by inverted microscope. Flow cytometry assays showed that artesunate significantly induced apoptosis in HGC27 cells. Af- ter treated with artesunate （20, 40, 80 mg/L）, the apoptosis rate of HGC27 cells was 11.5%, 21.4% and 36.6% respec- tively, which showed an obvious concentration-effect relationship, while the apoptosis rate of HGC27 cells was 2.2% in the control group. The relative activity of Caspase-3 of artesunate group was （0.19±0.02）, （0.25±0.04） and （0.31±0.03） respectively, which was significantly increased than the control group （0.11±0.02） （P 〈 0.05）. And the relative activity of Caspase-9 of artesunate group was （0.18±0.02）, （0.23±0.03） and （0.30±0.04）, which was significantly increased than the control group （0.10±0.02） （P 〈 0.05）. The data of Western blot showed that artesunate up-regulated RUNX-3 in a dose-dependent manner. Conclusion Artesunate can inhibit the proliferation of HGC27 cells and induce apoptosis, and the mechanism of artesunate on apoptosis may be related to the up-regulation of RUNX-3 expression, as well as the increase of relative activity of Caspase-3 and Caspase-9. Artesunate may be a promising antitumor agent for gastric cancer treatment.