β-二氢青蒿素-大黄素复合物对人胃癌细胞株SGC-7901作用的实验研究

Effects ofβ-dihydroartemisinin-emodin on experiment in human gastric cancer SGC-7901 cells

目的探讨β-二氢青蒿素-大黄素复合物能否诱导人胃癌SGC-7901细胞凋亡及相关作用机制。方法四唑盐(MTT)比色法检测β-二氢青蒿素-大黄素复合物对SGC-7901细胞的抗增殖及细胞毒作用、Hoechst33258荧光染色和HE染色观察细胞凋亡的形态学改变、AnnexinV-PI双染法检测凋亡、Western Blotting方法检测CyclinD1和Ki-67蛋白的表达变化。结果 MTT结果β-二氢青蒿素-大黄素复合物对SGC-7901细胞抑制作用显著,且呈浓度和时间依赖性(P<0.05);镜下观察可见明显的细胞凋亡形态;流式细胞仪检测结果显示,β-二氢青蒿素-大黄素复合物能诱导人胃癌SGC-7901细胞凋亡,且呈浓度依赖性(P<0.05);Western Blotting方法发现β-二氢青蒿素-大黄素复合物明显下调了CyclinD1和Ki-67的表达水平。结论β-二氢青蒿素-大黄素复合物可抑制人胃癌SGC-7901细胞的增殖,诱导其凋亡,其作用机制可能与细胞周期阻滞有关。

Objective To explore whetherβ-dihydroartemisinin-emodin could induce apoptosis and inhibit the proliferation of SGC-7901 cells,and if its mechanism is related to the blockage of cell cycle.Methods The anti-proliferation and cytotoxic effects ofβ-dihydroartemisinin-emodin on SGC-7901 cells were studied by MTT colorimetric assay;after being treated byβ-dihydroartemisinin-emodin and special stains,the morphological changes of SGC-7901 cells were observed with optical microscope and fluorescence microscope;the apoptosis inducement ofβ-dihydroartemisinin-emodin on SGC-7901 cells was measured with Annexin V PI flow cytometry;the expression of CyclinD1 and Ki-67 was detected with Western blotting.Resultsβ-dihydroartemisinin-emodin had the highest inhibition rate by MTT colorimetric assay.With the increasing of drug concentration and the extension of time,the anti-proliferation and cytotoxic effects ofβ-dihydroartemisinin-emodin on SGC-7901 cells strengthened,showing a dose-and time-dependent depression effect.By using Hoechst33258 and HE staining,cells were observed and compared at morphology level,it was clear that the apoptosis inducement ofβ-dihydroartemisinin-emodin on SGC-7901 cells.Annexin V PI flow cytometry:compared to control group,the apoptosis inducement ofβ-dihydroartemisinin-emodin on SGC-7901 cells objectively with statistically significance(P<0.05).Detection of Western Blotting:the expression of cell cycle related protein CyclinD1 and proliferating cell nuclear antigen Ki-67 detected in experimental group decreased.Conclusion The effect ofβ-dihydroartemisininemodin on the inhibition of proliferation in SGC-7901 cells had dose-effect relationship and induced apoptosis,its mechanism is related to the blockage of cell cycle.