论文部分内容阅读
目的探讨醌茜素对人胃癌细胞的凋亡作用及其分子机制。方法 MTT法检测醌茜素对3种人胃癌AGS、MKN-28及MKN-45细胞的杀伤作用;Annexin V-FITC/PI双染法、流式细胞术检测醌茜素诱导AGS细胞凋亡能力及细胞内活性氧簇(ROS)水平;蛋白质免疫印迹法检测细胞凋亡相关蛋白的表达量变化。结果醌茜素对3种胃癌细胞均具有明显的杀伤作用,且呈浓度依赖性(P<0.05)。经计算其IC50值分别为7.07μmol/L、22.55μmol/L及14.18μmol/L。醌茜素能诱导AGS细胞发生凋亡,且呈浓度依赖性(P<0.001),并能够促进细胞内活性氧水平升高(P<0.001)。当预处理ROS清除剂NAC后,能够明显抑制醌茜素诱导的细胞凋亡(P<0.001)。Western blotting结果显示促凋亡蛋白p-p38、p-JNK、Bad、cleaved-caspase-3及cleaved-PARP-1表达量增加(P<0.05),抗凋亡蛋白p-Akt、p-ERK及Bcl-2蛋白表达量减小(P<0.05)。结论醌茜素通过上调细胞内ROS水平,调控MAPK及Akt信号途径,进而诱导人胃癌AGS细胞发生凋亡。
Objective To investigate the apoptosis of human gastric cancer cells induced by quinizarin and its molecular mechanism. Methods MTT assay was used to detect the killing effect of quinizarin on AGS, MKN-28 and MKN-45 cells. Annexin V-FITC / PI double staining and flow cytometry were used to detect the apoptosis of AGS cells induced by quinizarin And intracellular reactive oxygen species (ROS). Western blotting was used to detect the expression of apoptosis-related proteins. Results Quinizarin had a significant killing effect on all three kinds of gastric cancer cells in a concentration-dependent manner (P <0.05). The calculated IC50 values were 7.07μmol / L, 22.55μmol / L and 14.18μmol / L, respectively. Quinizarin induced apoptosis in AGS cells in a dose-dependent manner (P <0.001), and increased the intracellular reactive oxygen species (P <0.001). When pretreated with ROS scavenger NAC, quinolizine-induced apoptosis was significantly inhibited (P <0.001). The results of Western blotting showed that the expressions of p-p38, p-JNK, Bad, cleaved-caspase-3 and cleaved-PARP- Bcl-2 protein expression decreased (P <0.05). Conclusion Quinizarin can induce the apoptosis of human gastric cancer cell line AGS by up-regulating ROS levels and regulating the MAPK and Akt signaling pathways.