Study of the effects in vitro of berberine combined with bortezomib on multiple myeloma cell line U266 cells
10.3760/cma.j.issn.0253-2727.2016.11.010
- VernacularTitle:小檗碱联合硼替佐米对多发性骨髓瘤细胞株U266细胞作用机制的体外研究
- Author:
Yin TIAN
1
;
Lihong LI
;
Guangzhong YANG
;
Wenming CHEN
Author Information
1. 100020,首都医科大学附属北京朝阳医院血液科
- Keywords:
Multiple myeloma;
Berberine;
Bortezomib;
In vitro
- From:
Chinese Journal of Hematology
2016;37(11):976-981
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of berberine in combination with bortezomib on proliferation and apoptosis of multiple myeloma (MM) cell line.Methods MM cell line U266 cells were treated with berberine and/or bortezomib.The effects of berberine and/or bortezomib on proliferation of cells were measured by methylthiazolyl tetrazolium bromide (MTT).Flow cytometric Annexin Ⅴ/PI double staining method was used to detect effect of either drug alone or in combination on apoptosis of MM cell line U266.ELISA was used to measure the expression of casepase-3,-8,-9 affected by the two drugs.Western blot was used to detect the expression of the apoptosis-related protein TRADD and FADD.King formula was used to determine if there was a synergistic effect of berberine in combination with bortezomib.Results ① Both berberine and bortezomib as single agent had dose-and time-dependent effects of proliferation inhibition on U266 cells.Berberine (20 μmol/L) and bortezomib (5 nmol/L) had a synergistic effect of proliferation inhibition (Q value:1.31-1.65).②The proportion of early stage apoptosis in both single agent groups and combination group significantly increased compared to control group (P< 0.05).Berberine and bortezomib had a synergistic effect on cell apoptosis (Q value after 6 h and 12 h were 0.896 and 1.197,respectively).③Berberine in combination with bortezomib significantly upregulated expressions of caspase-3,-8 and-9,which were statistically significant (P<0.05).④Berberine in combination with bortezomib significantly upregulated expressions of TRADD (0.91±0.01,0.70±0.01) and FADD (0.98±0.01,0.98±0.01) compared with control group (both P<0.05).Conclusion Berberine in combination with bortezomib had synergistic effects on proliferation inhibition and apoptosis,which were mediated by up-regulated levels of TRADD and FADD.