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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 study the effects of Saponins on the apoptosis of tumor cells and to study its mechanism. Methods: The proliferation of HepG2, A549 and SGC-7901 cells cultured in vitro was detected by MTT assay. The effect of drupe saponins on the cell cycle and apoptosis of HepG2 cells was studied by fluorescence staining and flow cytometry Western blotting was used to detect the expression of Bax, Bcl-2 and P53 proteins. Results: Saponin of HepG2 cells had a significant inhibitory effect. The IC50 of HepG2 cells at 24, 48 and 72 h were 13.99μg / ml, 11.78μg / ml and 10.01μg / ml respectively The results showed that the IC50 of 72h for A549 lung cancer cells was 23.68μg / ml, but had no significant inhibitory effect on the proliferation of SGC-7901 gastric cancer cells. The results of flow cytometry showed that the percentage of S phase cells in HepG2 cells was 28.81%, 34.96% and 46.57%, respectively, after treatment with orphanin at 10, 15 and 20μg / ml for 24 hours. The percentage of apoptotic cells 30.06%, 30.32% and 40.34% respectively. The results of Western blotting showed that the expression of Bax and P53 increased and the expression of Bcl-2 decreased with the increase of the concentration of saxapin. Conclusion: Sapogenin can inhibit the proliferation of HepG2 and A549 cells and induce the apoptosis of HepG2 cells. The mechanism may be related to the fact that Sapogenin up-regulates the expression of P53 protein, thus affecting the expression of Bax and Bcl-2, blocking the cells in S phase and Eventually lead to apoptosis.