葛根素糖苷生物合成及对乳腺癌细胞增殖抑制的影响

Biosynthesis of Puerarin Glycoside and Proliferation Inhibition Effect on Breast Cancer Cell

目的研究约氏不动杆菌G2菌株对葛根素糖苷生物合成的最佳转化条件,以及葛根素糖苷对人乳腺癌细胞株MDAMB-231的增殖抑制作用。方法考察生物合成体系中转化温度、pH和蔗糖浓度对转化率的影响;并以四甲基偶氮唑盐微量酶反应比色法(MTT法)观察不同浓度的葛根素及葛根素糖苷对MDA-MB-231细胞生长的影响。结果葛根素糖苷的最佳转化条件为温度30℃,pH 7.38,蔗糖浓度60g/L,在此条件反应24h,转化率为85%。MTT法显示,不同浓度的葛根素及其糖苷对MDA-MB-231细胞均有抑制作用,并随浓度升高而抑制作用增强;其中200μmol/L浓度葛根素及葛根素糖苷作用MDA-MB-231细胞48h后,其抑制率分别为61.91%和59.42%。结论约氏不动杆菌G2菌株可有效催化葛根素合成葛根素糖苷;葛根素及其糖苷均能明显抑制人乳腺癌细胞株MDA-MB-231的增殖。

OBJECTIVE To study the optimal transforming conditions for biosynthesis of puerarin glycoside by Acinetobacter johnsonii G2,and the proliferation inhibition effect of puerarin glycoside on human breast cancer cell line MDA-MB-231.METHODS Effect of temperature,pH and sucrose concentration on transformation rate in biosynthesis system were studied.Besides,effects of different concentrations of puerarin and puerarin glycoside on the proliferation of MDA-MB-231 cells through methyl thiazolyl tetrazolium trace enzyme reaction colorimetry （MTT assay） were investigated.RESULTS The best transformation condition of puerarin glycoside was 30 ℃,pH 7.38 and sucrose concentration of 60 g/L.In this condition,the transformation rate was 85 % after 24 h.MTT assays showed that different concentrations of puerarin and puerarin glycoside all could inhibit proliferation of MDA-MB-231 cells,and the inhibition effect strengthened with the concentration increased; among which the inhibition rate of 200μmol/L puerarin and puerarin glycoside on MDA-MB-231 cells were 61.91% and 59.42 % respectively after 48 h.CONCLUSION Acinetobacter johnsonii G2 can effectively catalyze synthesis of puerarin to puerarin glycoside; both puerarin and puerarin glycoside can significantly inhibit the proliferation of human breast cancer cell line MDA-MB -231.