浙贝母总生物碱对人肺腺癌A549/顺铂细胞耐药性的逆转作用(英文)  

M ultidrug resistance reversal activity of total alkaloid from Fritillaria thunbergii on cisplatin-resistant human lung adenocarcinom a A549/DDP cells

作　　者：李泽慧[1] 安超[2] 胡凯文[2] 周科华 段惠惠[1] 唐民科[1] 

机构地区：[1]北京中医药大学中药学院中药药理系 [2]北京中医药大学东方医院肿瘤科 [3]Department of Physical Therapy,Daemen College,Amherst 14226,USA

出　　处：《中国药理学与毒理学杂志》2013年第3期315-320,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家科技重大专项(2009ZX09103-346)~~

摘　　要：目的研究浙贝母总生物碱(TAF)对人肺腺癌A549/顺铂(DDP)细胞DDP耐药性的逆转作用。方法①离体实验:采用MTT法观察TAF(12.5～200mg·L-1)对A549和A549/DDP细胞的毒性作用;采用MTT法检测TAF 9mg·L-1对A549/DDP细胞耐药的逆转作用,同时设环孢菌素A(Cys A)1mg·L-1和汉防己甲素(Tet)1mg·L-1为阳性对照;实时荧光定量PCR检测A549/DDP细胞多药耐药基因1(MDR1)mRNA相对表达;Western蛋白免疫印迹法检测A549/DDP细胞P-糖蛋白(P-gp)相对表达。②在体实验:制备BALB/c裸鼠A549/DDP移植瘤模型,随机分为模型对照、DDP 5mg·kg-1、TAF2 mg·kg-1、DDP 5 mg·kg-1+TAF 0.5,1和2 mg·kg-1联用组,每两天ip给予DDP,每天ig给予TAF,持续13d,检测移植瘤体积和质量的变化。结果 TAF作用72 h抑制A549和A549/DDP细胞存活的IC50值分别为141±5和(298±22)mg·L-1;IC10值分别为15.3±1.9和(9.0±1.2)mg·L-1。DDP 0.01～100mg·L-1与TAF 9 mg·L-1合用后,DDP抑制A549/DDP细胞存活的IC50值由(14.06±3.72)mg·L-1降至(0.79±0.14)mg·L-1,抑制A549细胞存活的IC50值无明显变化;TAF对A549/DDP细胞DDP耐药性的逆转倍数为17.80倍,高于Cys A(10.16倍)和Tet(14.05倍)。TAF可明显降低A549/DDP细胞MDR1 mRNA及P-gp相对表达(P<0.01)。DDP 5mg·kg-1体内抑瘤率为49.9%,与TAF 2mg·kg-1合用后抑瘤率增至67.4%(P<0.01)。结论 TAF在体内外均可逆转A549/DDP细胞对DDP的耐药性,可降低MDR1 mRNA和P-gp蛋白表达。OBJECTIVE To explore the effect of total alkaloid from Fritillaria thunbergii（TAF） on reversing multidrug resistance（MDR） of human lung adenocarcinoma A549 /DDP cells.METHODS ① In vitro Cytoxicity and proliferation inhibitory rate of TAF（12.5-200 mg·L-1） was assessed by MTT method.TAF 9 mg·L-1 was used in subsequent reversal experiments.Cyclosporine A（Cys A） 1 mg·L-1 and tetrandrine（Tet） 1 mg·L-1 acted as positive control group.The amount of MDR1 mRNA and P-glycoprotein（P-gp） of A549 /DDP cells was measured by real time polymerase chain reaction and Western blotting,respectively.② In vivo BALB /c nude mice were used to establish an A549 /DDP tumor model.The mice were randomly divided into vehicle,DDP 5 mg·kg-1,TAF 2 mg·kg-1,DDP 5 mg·kg-1 plus TAF 0.5,1 and 2 mg·kg-1 groups.DDP was ip given every two days and TAF was ig given once a day.Tumor volume was measured every four days and tumor mass was detected after 13 d.RESULTS After incubation with TAF for 72 h,IC50 of TAF to A549 and A549 /DDP cells was 141 ± 5 and（298 ± 22） mg·L-1,respectively,and IC10 of TAF to A549 and A549 /DDP cells was 15.3 ± 1.9 and（9.0 ± 1.2） mg·L-1.IC50 of DDP without or with TAF 9 mg·L-1 on A549 /DDP cells was 14.06 ± 3.72 and（0.79 ± 0.14） mg·L-1,respectively,while there was no significant change in IC50 of A549 cells.TAF reversed DDP resistance of A549 /DDP cells with fold-reversal 17.80,which was higher than that of Cys A（10.16） and Tet（14.05）.Compared with A549 /DDP cell vehicle group,MDR1 mRNA and P-gp expression in A549 /DDP cells was decreased by TAF（P 0.01）.The tumor inhibitory rate of DDP 5 mg·kg-1 in vivo was 49.9%.The combination of DDP 5 mg·kg-1 and TAF 2 mg·kg-1 increased the tumor inhibition rate to 67.4%.CONCLUSION TAF enhances MDR reversal effect of DDP on A549 /DDP cells in vitro and in vivo,and down-regulates MDR1 mRNA and P-gp expression in A549 / DDP cells.

关 键 词：总生物碱 浙贝母 多药耐药性 P-糖蛋白 

分 类 号：R734.2[医药卫生—肿瘤]