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目的:研究纳米雄黄对肺癌A549细胞及其肿瘤干细胞(cancer stem cells,CSC)的凋亡诱导作用。方法:采用机械研磨法制备纳米雄黄。以肺癌A549细胞为靶细胞,采用MTT比色法检测细胞的增殖活性;Annexin V/PI双染色法检测A549细胞及其CSC的凋亡,流式细胞术检测P-糖蛋白(P-glycoprotein,P-gp)和乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)表达、Caspase-3活性及群体细胞中CSC含量。结果:纳米雄黄显著抑制A549细胞的增殖,50μg/ml和100μg/ml的纳米雄黄处理48h后,细胞凋亡率分别为12.53%和69.19%;作用24h后,细胞中活化caspase-3由对照的(2.25±0.17)%增高到(3.84±0.63)%和(7.35±0.33)%。20μg/ml、50μg/ml和100μg/ml纳米雄黄处理48h,细胞中CSC的相对含量有所增高,同时CSCs的凋亡率明显增高,分别为(4.28±0.42)%、(9.17±1.11)%和(30.71±2.82)%,但低于群体细胞。纳米雄黄作用后A549细胞P-gp和BCRP表达变化不明显。结论:纳米雄黄可有效诱导肺癌A549细胞及其肿瘤干细胞发生凋亡。
AIM: To investigate the apoptosis-inducing effect of nano-realgar on lung cancer A549 cells and its cancer stem cells (CSCs). Methods: Nano realgar was prepared by mechanical grinding method. The proliferation of A549 cells was detected by MTT assay. The apoptosis of A549 cells and CSCs was detected by Annexin V / PI double staining. The expressions of P-glycoprotein (P-glycoprotein, P-gp) and breast cancer resistance protein (BCRP) expression, Caspase-3 activity and CSC content in population cells. Results: Nano-realgar significantly inhibited the proliferation of A549 cells. After treated with 50μg / ml and 100μg / ml nano-realgar for 48h, the apoptotic rates were 12.53% and 69.19%, respectively. After 24h, the activated caspase-3 (2.25 ± 0.17)% to (3.84 ± 0.63)% and (7.35 ± 0.33)%, respectively. The relative content of CSC increased in the cells treated with 20μg / ml, 50μg / ml and 100μg / ml nano realgar for 48h, and the apoptotic rates of CSCs were significantly higher (4.28 ± 0.42%, 9.17 ± 1.11% And (30.71 ± 2.82)%, respectively, but lower than the population of cells. Nano-realgar A549 cells after P-gp and BCRP expression did not change significantly. Conclusion: Nano-realgar can effectively induce the apoptosis of lung cancer A549 cells and cancer stem cells.