Yin Jun-Qiang, Shen Jing-Nan, Su Wei-Wei, Wang Jin, Huang Gang, Jin Song, Guo Qian-Chen, Zou Chang-Ye, Li Hao-Miao, Li Fo-Bao
Department of Orthopaedic Oncology, First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, China.
Acta Pharmacol Sin. 2007 May;28(5):712-20. doi: 10.1111/j.1745-7254.2007.00559.x.
To investigate the antiproliferative activity and apoptosis-inducing effects of bufalin on human osteosarcoma cell lines.
U-2OS and U-2OS methotrexate (MTX) 300-resistant cell lines were treated with bufalin. Cell viability was assessed using the MTT assay. Cell-cycle status, apoptosis-inducing effects, and the expression of apoptosis-related proteins were evaluated by flow cytometry, fluorescent staining, DNA fragmentation assays, and Western blotting. The effect of bufalin on dihydrofolate reductase (DHFR) expression was studied by RTPCR and Western blotting.
Bufalin inhibited cell growth in both U-2OS and U-2OS MTX300 cells. The induction of G2/M cell-cycle arrest was also seen in the cells treated with bufalin. The induction of apoptosis by bufalin was confirmed by increased expression of the tumor suppressor protein p53 and the increased ratio of the Bax/Bcl-2 proteins. Bufalin induced apoptosis to the same extent in both cell lines without regard to DHFR levels in the cells.
Bufalin inhibited the growth of and induced apoptosis in both MTX-sensitive and MTX-resistant human osteosarcoma U-2OS cells. The apoptosis-inducing effect of bufalin was not influenced by the presence of high levels of the DHFR protein.
研究蟾毒灵对人骨肉瘤细胞系的抗增殖活性和诱导凋亡作用。
用蟾毒灵处理U-2OS和U-2OS甲氨蝶呤（MTX）300耐药细胞系。采用MTT法评估细胞活力。通过流式细胞术、荧光染色、DNA片段化分析和蛋白质免疫印迹法评估细胞周期状态、诱导凋亡作用及凋亡相关蛋白的表达。通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法研究蟾毒灵对二氢叶酸还原酶（DHFR）表达的影响。
蟾毒灵抑制U-2OS和U-2OS MTX300细胞的生长。在用蟾毒灵处理的细胞中也观察到G2/M期细胞周期阻滞。肿瘤抑制蛋白p53表达增加以及Bax/Bcl-2蛋白比例增加证实了蟾毒灵诱导细胞凋亡。无论细胞中DHFR水平如何，蟾毒灵在两种细胞系中诱导凋亡的程度相同。
蟾毒灵抑制MTX敏感和MTX耐药的人骨肉瘤U-2OS细胞的生长并诱导其凋亡。蟾毒灵的诱导凋亡作用不受高水平DHFR蛋白存在的影响。
