DMSO对悬浮培养的东北红豆杉细胞凋亡和紫杉醇合成的影响

Effect of DMSO on Apoptosis and Taxol Biosynthesis in Suspension Cultured Taxus cuspidata Cells

研究了不同浓度的DMSO对悬浮培养的东北红豆杉 (Taxuscuspidata)细胞的增殖能力、细胞活性以及紫杉醇合成和释放等方面的影响 ,同时应用荧光指示剂双染法检测了细胞凋亡的发生情况。结果显示2 %的DMSO处理能显著降低细胞活性 ,抑制细胞的增殖能力 ,使细胞核内DNA含量减少 ,培养中、后期在荧光显微镜下可见部分细胞核出现典型的凋亡形态 ,同时伴有紫杉醇产量的明显增加 ;对照组及 1%以下浓度组未出现上述改变。结果表明一定浓度的DMSO能诱导细胞凋亡 。

The effects of different concentrations of DMSO on cell proliferation, cell viability and secondary product biosynthesis of Taxus cuspidata in suspension cultured cells were studied. Meanwhile, apoptosis was detected with fluorescence dyes PI and Hoechst 33342 used for staining. The results showed that 2% DMSO could decrease the cell viability significantly(Fig.3), suppress cell proliferation(Fig.1), and reduce the DNA content in the nucleus(Fig.2). Some of the cells treated with 2% DMSO showed typical apoptotic morphological changes under fluorescence microscope from the 8th to the 27th day of culture(Plate I b,c,d). In the same time, the yield of taxol was enhanced evidently(Fig.5). These changes in the above description were not observed in the control group and the groups treated by DMSO of concentrations 0.5% and 1%. These findings show that DMSO at a certain concentration induces apoptosis and promotes taxol biosynthesis by Taxus cuspidata cells in suspension culture.