研究刺五加多糖（Acanthopanax senticosus polysaccharide,ASPS）诱导H446细胞凋亡及其可能的作用机制。采用MTT法检测ASPS对小细胞肺癌H446细胞增殖的抑制作用;Hoechst33258染色和流式细胞技术检测经ASPS处理后H446细胞凋亡的形态特征及凋亡率的变化;Western印迹方法检测凋亡相关基因bax、bcl-2、p53表达的变化。MTT分析表明,ASPS作用48h后可明显抑制H446细胞的增殖,半数抑制浓度（IC50值）为476.36μg/ml;Hoechst染色结果：H446细胞在ASPS诱导下出现典型的凋亡形态;流式细胞术检测结果显示：对照组及浓度为240、480、960μg/ml药物处理组凋亡率分别是（5.02±0.4）%、（11.12±1.8）%、（19.89±2.5）%、（22.54±1.8）%;Western印迹显示：在ASPS的诱导下bax、p53的表达量提高,而bcl-2的表达量下降。研究表明,ASPS对H446细胞增殖有抑制作用,并能促进其凋亡;ASPS通过上调bax、p53表达,下调bcl-2表达促进H446细胞凋亡。
To investigate Acanthopanax senticosus polysaccharide （ASPS） induced apoptosis in small cell lung cancer cell lines H446 and its potential mechanism. MTT assay was used to determine the inhibitory rate of ASPS with different concentrations on the proliferation of H446 cells. The cell apoptosis induced by ASPS was detected with Hoechst 33258 dye and flow cytometry （FCM）. Western blot was used to detect the changes of bax, bcl-2 and p53 in protein levels in H446 cells. MTT assay showed that the proliferation of H446 cells were markedly inhibited after treatment with ASPS for 48 h; IC50 was 476.36 μg/ml; Typical morphological changes of apoptosis were observed in H446 cells with Hoechst staining after induced by ASPS. FCM analysis revealed the apoptotic rates of H446 cells treated by 0, 240, 480, 960 μg/ml ASPS were （5.02±0.4）%, （11.12±1.8）%, （19.89±2.5）%, （22.54± 1.8）% respectively. Western blot results indicated that the expression of bax and p53 were increased but bcl-2 was decreased by ASPS. ASPS could inhibit the proliferation of H446 cells and induce part of cells to apoptosis. The activation of bax and p53 and the suppression of bc1-2 may contribute to the apoptosis mechanism.
Chinese Journal of Cell Biology