竹节参皂苷对顺铂耐药肺癌细胞敏感性的影响及机制研究

Study on the Effect and Mechanism of Saponins from Panax Japonicus in Inhibiting Cisplatin Resistant Lung Cancer Cells

关键词:  竹节参皂苷  顺铂  耐药

Key words:  saponins from Panax japonicus  cisplatin  drug resistant

摘要:

目的 研究竹节参皂苷对顺铂耐药肺癌细胞敏感性的影响及其潜在作用机制。方法 通过递增顺铂浓度诱导肺癌A549细胞耐药，测定细胞存活率，SRB显色法测定药物敏感性，Annexin V-FITC/PI双染测定细胞凋亡，ELISA测定Caspase 3和Caspase 8水平，Western blotting测定细胞中相关蛋白表达情况。结果 经长期诱导后，A549细胞对顺铂的耐受性增加14.79倍，且较稳定。与顺铂组比较，联合应用竹节参皂苷和顺铂可显著增加A549细胞对顺铂的敏感性，增加细胞内Caspase 3和Caspase 8水平，促进细胞凋亡，同时抑制MDR1、P-gp、Bcl-2和p-Akt等蛋白表达，促进Bax蛋白表达。结论 竹节参皂苷可增加顺铂耐药细胞株对顺铂的敏感性，增强顺铂的作用效果，其作用机制可能与抑制耐药相关蛋白MDR1、P-gp和Akt活性相关。

Abstract:

OBJECTIVE To study the effect of saponins from Panax japonicus on the sensitivity of cisplatin resistant lung cancer cell and further discuss its potential mechanism. METHODS Drug resistance of lung cancer A549 cells was induced by increasing concentration of cisplatin, and then cell viability was determined, the drug sensitivity was determined by SRB colorimetry, apoptosis was detected by Annexin V-FITC/PI double staining assay, Caspase 3 and Caspase 8 levels was measured by ELISA and the expression of related proteins was determined by Western blotting. RESULTS After long-term induction, the tolerance of A549 cells to cisplatin was increased by 14.79 times and was relatively stable. Compared with cisplatin treatment group, the combination of saponins from Panax japonicus and cisplatin significantly increased the sensitivity of A549 cells to cisplatin, increased the intracellular Caspase 3 and Caspase 8 levels and promoted apoptosis. At the same time, the expression of MDR1, P-gp, Bcl-2 and p-Akt was inhibited, and the expression of Bax protein was promoted. CONCLUSION Saponins from Panax japonicus can increase the sensitivity of cisplatin-resistant cell lines to cisplatin and enhance the effect of cisplatin, and the possible mechanism may be through inhibiting the activity of drug-resistant related proteins MDR1, P-gp and Akt.