【摘要】 目的探讨千金藤碱抑制肺癌细胞A549生长、迁移和侵袭的作用以及对血红素加氧酶-1(heme oxy-genase-1,HO-1)和表皮生长因子受体(epidermal growth factor receptor,EGFR)基因表达的影响。方法甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)比色法检测千金藤碱对肺癌细胞的细胞毒性,观察其对A549细胞生长的抑制作用,并用倒置显微镜观察千金藤碱处理不同时间内A549细胞的形态变化;Transwell和侵袭实验检测A549细胞迁移和侵袭能力;RT-PCR和Western blot法分别检测A549细胞内HO-1和EGFR基因的mRNA和蛋白表达水平。结果千金藤碱能够抑制肺癌细胞A549细胞的生长,细胞活力随着作用浓度的增加和时间的延长明显降低;10g/ml千金藤碱处理24h后,A549细胞发生体积变小、皱缩变圆和染色体固缩等现象;与对照组相比,千金藤碱(>5.0g/ml)可使穿过隔膜和穿过基质胶的A549细胞数明显减少(P<0.05)。10.0g/ml千金藤碱明显降低A549细胞中HO-1和EGFR基因的mR-NA和蛋白表达水平(P<0.05)。结论千金藤碱可抑制肺癌细胞的增殖,降低A549细胞的迁移和侵袭能力,下调A549细胞的HO-1和EGFR基因的表达水平,且作用效果具有一定的时间和浓度依赖性。更多还原

【Abstract】 Objective To investigate the inhibitory effects of cepharanthine(CEP) on the growth,migration,and invasion of human lung cancer cell line A549and the gene expression of heme oxygenase-1(HO-1) and epidermal growth factor receptor(EGFR) in A549cells.Methods The viability of A549cells treated with CEP was determined by methyl thiazolyl tetrazolium assay,and the morphological changes of A549 cells at different points of treatment with CEP were observed under an inverted microscope.The migration and invasion of A549cells were evaluated using Transwell chambers and Matrigel-coated Transwell chambers,respectively.The mRNA and protein expression levels of HO-1and EGFR in A549cells were measured by RT-PCR and western blot,respectively.Results CEP significantly inhibited the growth of A549 cells,and the cell viability decreased significantly as the concentration of CEP increased and the treatment proceeded.After being treated with 10g/ml CEP for 24h,A549cells became smaller,shrunken,and rounded and showed chromatin condensation.There were significantly fewer cells that crossed the Transwell membrane and Matrigel in the A549cells treated with CEP(>5.0g/ml) than in controls(P< 0.05).CEP(10.0g/ml) significantly decreased the mRNA and protein expression levels of HO-1and EGFR in A549cells(P<0.05).Conclusion CEP can inhibit the growth,migration,and invasion of A549 cells and downregulate the mRNA and protein expression of HO-1and EGFR in A549cells.The effects of CEP display certain time and concentration dependence.更多还原