论文部分内容阅读
目的:通过体外实验研究Protobioside对人肝癌细胞株HepG2的细胞增殖抑制作用,并进一步研究其作用机制。方法:采用噻唑兰(MTT)比色法测定Protobioside对3种肿瘤细胞生长增殖的影响;采用Hoechst33528染色观察Protobioside对HepG2细胞凋亡的影响;采用流式细胞仪技术检测Protobioside对HepG2细胞周期的影响;采用蛋白质电泳技术检测Protobio-side对HepG2细胞的细胞周期相关蛋白及凋亡相关蛋白表达的影响。结果:Protobioside对HepG2细胞的生长抑制作用最强,IC50为20μmol.L-1;形态学观察可见Protobioside处理36 h时细胞核发生皱缩扭曲、胞膜完整。细胞周期分析显示,Protobio-side将HepG2细胞抑制在G2/M期,具有良好的时效、量效关系;蛋白质电泳结果表明Protobioside下调了周期蛋白Cyclin B1、上调了促凋亡蛋白Bax、下调了抑凋亡蛋白Bcl-2。结论:Protobioside通过下调周期蛋白Cyclin B1将人肝癌细胞HepG2细胞阻滞在G2/M期,通过上调促凋亡蛋白Bax、下调抑凋亡蛋白Bcl-2诱导HepG2细胞凋亡,从而抑制HepG2细胞的增殖。
OBJECTIVE: To study the inhibitory effect of Protobioside on HepG2 cell proliferation in vitro and to investigate its mechanism. Methods: The effects of Protobioside on the growth and proliferation of three tumor cells were determined by MTT colorimetric assay. The effect of Protobioside on the apoptosis of HepG2 cells was observed by Hoechst33528 staining. The effect of Protobioside on the cell cycle of HepG2 was detected by flow cytometry The effect of Protobio-side on the expression of cell cycle related proteins and apoptosis related proteins in HepG2 cells was detected by protein electrophoresis. RESULTS: Protobioside had the strongest growth inhibitory effect on HepG2 cells with an IC50 of 20 μmol·L-1. Morphological observation showed that the nuclei of the cells were wrinkled and twisted when Protobioside was treated for 36 h, and the cell membrane was intact. Cell cycle analysis showed that Protobio-side inhibited HepG2 cells in G2 / M phase with good aging and dose-effect relationship. The results of protein electrophoresis showed that Protobioside down-regulated cyclin B1, up-regulated pro-apoptotic protein Bax and down-regulated Apoptosis protein Bcl-2. CONCLUSIONS: Protobioside blocks HepG2 cells in G2 / M phase by down-regulating cyclin B1, induces apoptosis of HepG2 cells by up-regulating pro-apoptotic protein Bax and down-regulating apoptosis-inducing protein Bcl-2, thereby inhibiting HepG2 cells proliferation.