开口箭皂苷诱导肿瘤细胞凋亡研究  

Anti-proliferative effects of saponins from Tupistra chinensis Baker. in vitro

作　　者：谢锦艳[1] 张东东[1] 李玉泽[1] 李真[1] 宋蓓[1] 乔蓉霞 刘海静 宋小妹[1] 

机构地区：[1]陕西中医学院药学院,咸阳712046 [2]陕西省食品药品检验所,西安710065

出　　处：《中药药理与临床》2014年第3期82-86,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：陕西省科技统筹创新计划项目(2013KTCQ03-14)

摘　　要：目的:研究开口箭皂苷对肿瘤细胞凋亡的影响,并对其机制进行研究。方法:采用MTT法检测开口箭皂苷对体外培养的肿瘤细胞系HepG2、A549和SGC-7901增殖的影响,利用荧光染色技术及流式细胞术研究开口箭皂苷对HepG2细胞周期和细胞凋亡的影响,利用Western blotting技术检测Bax、Bcl-2、P53蛋白的表达情况。结果:开口箭皂苷对HepG2肝癌细胞具有显著的抑制作用,其24、48及72 h对HepG2细胞的半数抑制浓度(IC50)分别为13.99μg/ml、11.78μg/ml和10.01μg/ml,且呈一定的量效关系和时效关系;对A549肺癌细胞的72h的IC50为23.68μg/ml,而对SGC-7901胃癌细胞的增殖无明显抑制作用。流式细胞仪检测结果显示,HepG2细胞经浓度为10、15和20μg/ml的开口箭皂苷处理24h后,S期细胞的比例分别为28.81%、34.96%、46.57%;凋亡细胞的比例分别为30.06%、30.32%和40.34%。Western blotting结果显示,随着开口箭皂苷浓度的增加,Bax、P53蛋白的表达量增加,而Bcl-2的表达量减少。结论:开口箭皂苷可抑制HepG2和A549细胞的增殖并诱导HepG2细胞凋亡,其机制可能与开口箭皂苷上调P53蛋白表达,从而影响Bax和Bcl-2表达,使细胞阻滞于S期,并最终导致细胞凋亡。Objective： To look for the apoptosis of saponins from Tupistra chinensis Baker. by macroporous resin and its mechenism. Methods：MTT method was performed to detect the inhibition of saponins of Tupistra chinensis Baker upon to HepG2,A549 and SGC-7901 cell lines.DAPI staining and flow cytometry（ FCM） are used to observe the changes of cell cycle and apoptosis of HepG2 cell induced by the saponins of Tupistra chinensis Baker. The effects on expression of Bax,Bcl-2 and P53 proteins in HepG2 cells were detected by Western blot. Results：Saponins of Tupistra chinensis Baker has obvious inhibitory effect on HepG2 cells after 24h,48h and 72h. The IC50 of HepG2 cell treated by saponins of Tupistra chinensis Baker for 24h,48h and 72h were 13. 99,11. 78 and 10. 01 μg /ml,respectively. The results showed the rate of apoptosis induced by saponins of Tupistra chinensis Baker increase in a dose-dependent and time-dependent manner. Saponins of Tupistra chinensis Baker has definite inhibitory effect on A549 cells but has no obvious inhibitory effect of SGC7901 cells. The IC50 of A549 cells is23. 68 μg /ml after 72h. FCM assay indicated that the percentage of the cell cycles arresting at S phase after treatment with 10,15 and 20μg/ml saponins of Tupistra chinensis Baker for 24 h were increased to 28. 81%,34. 96% and 46. 57% from 23. 75%. By western blot analysis,we found that expression of Bax,P53 proteins in HepG2 cells was up-regulated with varying concentration of saponins of Tupistra chinensis Baker,and the Bcl-2 proteins was down-regulated. Conclusion： Saponins of Tupistra chinensis Baker can inhibit HepG2 and A549 cells proliferation,and induce HepG2 cell apoptosis,which has a obvious anti-proliferative activity in vitro. The mechanism may be related to expression of Bax and Bcl-2 protein by up-regulated expression of P53 protein,thus block cell cycle in S phase,and eventually lead to apoptosis.

关 键 词：开口箭 人肝癌细胞株(HepG2) 人肺腺癌细胞(A549) 人胃癌细胞株(SGC7901) 凋亡 

分 类 号：R285[医药卫生—中药学]