牛蒡子苷元对人食管癌细胞增殖和凋亡的影响及机制研究

Study of effects and mechanisms of arctigenin on cell proliferation and apoptosis of human esophageal cancer cells

目的:观察牛蒡子苷元(arctigenin,ARG)对体外培养的人食管癌细胞(esophageal cancer-1 cells,EC-1)增殖和凋亡的影响。方法:将不同浓度的ARG作用于EC-1,噻唑蓝(methyl thiazolyl tetrazolium,MTT)实验检测细胞增殖抑制率、流式细胞术测定细胞周期变化,免疫组化法测定增殖细胞核抗原(Proliferating CellNuclear Antigen,PCNA)表达水平;将不同浓度的ARG作用于EC-1,流式细胞术检测细胞凋亡率、琼脂糖凝胶电泳和DNA缺口末端标记法(terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labelingtechnique,TUNEL)来检测细胞凋亡,免疫组化检测Bcl-2、Bax凋亡基因表达水平。结果:与对照组比较,ARG对EC-1细胞生长有抑制作用,随着时间/浓度的增加抑制作用逐渐增强(P<0.05);ARG可抑制EC-1由G0/G1向S期转换,G0/G1期细胞百分率较对照组显著增加,S期细胞百分率显著降低;随ARG浓度增加EC-1内PCNA表达水平逐渐降低。随ARG浓度增加,EC-1细胞凋亡率和凋亡指数显著增加(P<0.05);随ARG浓度增加,Bcl-2表达逐渐减弱,Bax表达逐渐增强(P<0.05)。结论:ARG有抑制EC-1增殖,下调EC-1内PCNA表达水平,阻滞细胞周期的作用。ARG可明显诱导EC-1细胞凋亡可能与ARG调控Bcl-2,Bax基因表达有关。

Objective： To investigate the effects of proliferation and apoptosis induced by ARG in human esophageal cancer-1 cells.Methods： EC-1 cells were treated with different concentration ARG.Proliferation of EC-1 cell inhibited by ARG were assessed by MTT.The cell cycle was measured using flow cytometry（FCM）,and the level of proliferating cell nuclear antigen（PCNA） with immunohistochemistry（IHC）.Apoptosis rate of EC-1 cell were assessed by flow cytomeu-y（FCM）.the changes of apoptosis of EC-1 cells were detected by TUNEL.The expression alteration of protein of Bcl-2 and Bax genes were detected by immunocytochemistry technique.Results： ARG could inhibit the growth and significantly suppressed expression of PCNA of EC-1 cells in a dose/time dependent manner（P0.05）.G0/G1 to S phase transition was blocked after treated with ARG,the percentage of G1 phase of the cell cycle was significantly increased,whereas the percentage of S phase was remarkably decreased.ARG could inhibit the growth of EC-1 cells in a dose/time dependent manner（P0.05）.TUNEL showed the ratio of apoptotic rate of EC-1 cells increased（P0.05）.Immunocytochemistry technique showed that the Bax protein increased gradually,while the Bcl-2 protein decreased gradually（P0.05）.Conclusion： The ARG could inhibit the growth and expression of PCNA,and retarded cell cycle of EC-1.ARG can induce apoptosis of EC-1 cells,the pathway that ARG induce apoptosis of EC-1 cells may be through regulation of protein expression of Bcl-2 gene and Bax gene.