小檗碱增加乳腺癌细胞对阿霉素药物敏感作用的实验研究

Experimental study on the effect of berberine on increasing the sensitivity of breast cancer cells to adriamycin

目的:观察小檗碱联合阿霉素对三阴性乳腺癌MDA-MB-231细胞株增殖的影响及潜在机制。方法:培养三阴性乳腺癌MDA-MB-231细胞株;CCK-8检测不同浓度阿霉素和联用小檗碱后细胞的存活率变化;Western blot检测细胞内p300、hSIRT-2、H3K56ac蛋白表达情况;细胞划痕实验、侵袭小室法检测药物对MDA-MB-231细胞迁移和侵袭能力的影响。结果:阿霉素对细胞增殖的抑制作用呈浓度依赖性,相同浓度阿霉素联合终浓度80μmol/L盐酸小檗碱对细胞的增殖抑制率均明显高于单独使用阿霉素(P<0.05)。p300、hSIRT-2、H3K56ac三种蛋白在对照组及加药组中均有表达,hSIRT-2蛋白在加药后含量降低,p300和H3K56ac蛋白的含量仅在含小檗碱组的细胞中增多,差异有统计学意义(P<0.05)。细胞侵袭和迁移实验显示小檗碱可以有效增强阿霉素对乳腺癌MDA-MB-231细胞三维迁移能力的抑制作用。结论:小檗碱可协同阿霉素抑制MDA-MB-231细胞增殖,共同发挥抗肿瘤效果,作用机制可能与p300、hSIRT-2、H3K56ac蛋白有关。

Objective:To observe the effect of berberine combined with adriamycin on the proliferation of MDA-MB-231 cell line in triple negative breast cancer and its potential mechanism.Methods:The MDA-MB-231 cell line of triple negative breast cancer was cultured and the changes of cell morphology were observed before and after the treatment.CCK-8 was used to detect the changes of survival rate of adriamycin and berberine in different concentrations.The expressions of p300,hSIRT-2 and H3K56ac proteins in the cells were detected by Western blot.The effects of drugs on the migration and invasion of MDA-MB-231 cells were examined by scratch test and invasion chamber method.Results:The inhibitory effect of adriamycin on cell proliferation was concentration-dependent.The inhibition rate of adriamycin combined with 80μmol/L berberine hydrochloride at the same concentration was significantly higher than that of adriamycin alone(P<0.05).p300,hSIRT-2 and H3K56ac proteins were all expressed in the control group and the treatment group,and the content of hSIRT-2 protein decreased after adding drugs.The content of p300 and H3K56ac protein only increased in berberine-containing cells,and the difference was statistically significant(P<0.05).Cell invasion and migration assay showed that berberine could effectively enhance the inhibitory effect of adriamycin on three-dimensional migration of breast cancer MDA-MB-231 cells.Conclusion:Berberine can cooperate with adriamycin to inhibit the proliferation of MDA-MB-231 cells and exert its anti-tumor effect,and the mechanism may be related to p300,hSIRT-2 and H3K56ac proteins.