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
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摘要: 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MDA-MB-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60 g/L，在此条件反应24 h，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200 μmol/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。Abstract: 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.
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