新月大戟提取液诱导人胃癌细胞SGC7901凋亡的研究

Study on the induction of apoptosis in human gastric carcinoma SGC7901 cells by Euphorbia lunulata extracts

目的研究新月大戟提取物诱导人胃癌SGC7901细胞的凋亡,并初步探索其可能的作用机制。方法用不同稀释度的新月大戟提取液处理SGC7901胃癌细胞,MTT法检验肿瘤细胞生长抑制情况;用HO与PI染色后,在荧光显微镜下观察凋亡细胞的核形态改变;透射电子显微镜下观察凋亡细胞的形态学改变;用流式细胞术检测细胞凋亡率和凋亡峰;Western blot检测细胞色素C的亚细胞分布。结果新月大戟提取液对SGC7901胃癌细胞的增殖有抑制作用,并呈时间和浓度依赖性;荧光显微镜下可观察到典型的凋亡细胞的核形态学改变;透射电子显微镜下,可观察到出泡或冒泡现象。用流式细胞术可检测到细胞凋亡率的增加和凋亡峰(亚G1峰)。新月大戟处理过的SGC7901细胞,细胞色素C的亚细胞分布发生了变化:线粒体内细胞色素C明显减少,而胞浆内的细胞色素C明显增强。结论新月大戟提取液具有抑制人胃癌SGC7901细胞增殖和诱导细胞凋亡的作用,其作用机制可能是通过线粒体通路或内源性凋亡的途径。

OBJECTIVE To study the induction of apoptosis in gastric carcinoma cell line SGC7901 by the natural extracts of Euphorbia lunulata and preliminarily explore the action mechanism. METHODS SGC7901 cells were treated with different concentra- tions of Euphorbia lunulata extracts and MTT test was conducted to measure cell viability. After stained with Hoechst 33258 （HO） and propidium iodide （PI）, the apoptotic cell nucleic morphology was observed with fluorescent microscope. Morphological changes of apop- totic cell was detected with Transmission electron microscope（TEM） Flow eytometry （FCM） was performed to detect the apoptotic rate and apoptotic peak. Western blot was employed to examine the subcellular localization of cytochrome C. RESULTS The extracts of Euphorbia lunulata induced SGC7901 cells to undergo apoptosis, which was time and concentration dependent. HO and PI staining and fluorescent microscopy showed the typical nucleic morphology changes of apoptotic cells. Through TEM ,foam or bubbling was observed. FMC examination showed an increased apoptotic rate and the typical apoptotic peak （ sub - G1 peak）. SGC7901 cells treated with Euphorbia iunulata extracts showed altered distribution of cytochrome C： mitochondrial cytochrome decreased while cytoplasmic cytochrome increased. CONCLUSION The results indicate that Euphorbia lunulata extracts could inhibit cell proliferation and induce apoptosis in SGC7901 cells and preliminarily revealed that the mechanism of action is through activating the mitochondrial pathway or the internal apoptotic pathway.