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目的:研究益气养肺方对肺癌耐药细胞体外生长和多药耐药相关蛋白(MRP)表达的影响,探讨益气养肺方干预肺癌耐药的线粒体及半胱天冬酶机制。方法:采用体外培养肺癌耐药细胞株SW 1573/2R120,分别以益气养肺方和阿霉素作用于肺癌细胞,通过MTT法检测细胞毒作用,流式细胞仪检测MRP蛋白表达,共聚焦显微镜测定线粒体膜电位和胞内钙离子(Ca2+)浓度,酶标法测定半胱天冬酶Caspase-3和Caspase-8活性。结果:与正常对照组相比,益气养肺方组细胞抑制率明显增加,MRP蛋白表达降低,线粒体膜电位和胞内Ca2+水平下降,Caspase-3和Caspase-8活性增加。结论:益气养肺方具有干预和逆转肺癌耐药的作用,其分子机制可能与下调线粒体膜电位水平和上调Caspase-3、Caspase-8活性有关。
Objective: To investigate the effect of Yiqi Yangfei Recipe on the growth of lung cancer cell lines in vitro and the expression of multidrug resistance-related protein (MRP), and explore the mechanism of Yiqi Yangfei Recipe in intervening mitochondrial and caspase-resistant lung cancer cells. Methods: Lung cancer cell line SW1573 / 2R120 was cultured in vitro. The lung cancer cells were treated with Yiqi Yangfei Recipe and doxorubicin respectively. The cytotoxicity was detected by MTT assay. MRP protein expression was detected by flow cytometry and confocal Mitochondrial membrane potential and intracellular calcium (Ca2 +) concentration were measured with a microscope. Caspase-3 and Caspase-8 activity were determined by enzyme-linked immunosorbent assay. RESULTS: Compared with the normal control group, the inhibition rate of Yiqi Yangfei Fang group was significantly increased, the expression of MRP protein was decreased, the mitochondrial membrane potential and intracellular Ca2 + level were decreased, and the activities of Caspase-3 and Caspase-8 were increased. Conclusion: Yiqi Yangfei Recipe can intervene and reverse the drug resistance of lung cancer. The molecular mechanism may be related to the down-regulation of mitochondrial membrane potential and up-regulation of Caspase-3 and Caspase-8 activity.