从PTEN-PI3K-AKT信号通路探讨补肾疏肝方含药血清抑制人肺腺癌A549细胞增殖和转移的机制  

Inhibiting Effect and Mechanism of Drug-contained Serum of Bushen Shugan Formula on A549 Cells Proliferation and Metastasis by PTEN-PI3K-AKT Pathway

作　　者：尚亚丽[1] 王淑玲[2] 

机构地区：[1]郑州大学第一附属医院,郑州450052 [2]郑州大学基础医学中西医结合研究所,郑州450052

出　　处：《中国实验方剂学杂志》2015年第10期153-157,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省科技厅重点攻关项目(311700531510)

摘　　要：目的:通过研究补肾疏肝方含药血清对PTEN-PI3K-AKT信号通路的影响,探讨补肾疏肝方抑制肺腺癌细胞增殖和转移作用机制。方法:大鼠60只,随机分为6组,每组10只,分别为生理盐水组(NS为2 m L),顺铂组(8 mg·kg-1),补肾疏肝方低剂量组(15 g·kg-1),补肾疏肝方高剂量组(30 g·kg-1),联合顺铂低剂量组(15 g·kg-1+8 mg·kg-1),联合顺铂高剂量组(30 g·kg-1+8 mg·kg-1),每只大鼠ig,2 m L,每天2次,共5 d。制备药物血清:经药物作用后的大鼠,经腹主动取血,分离血清,灭活,过滤,冻存。MTT法检测补肾疏肝方含药血清对A549细胞增殖的影响。三维细胞培养观察并计数各组细胞形成管状结构数量。RT-PCR检测含药血清对A549肺腺癌细胞AKT-1,Cyclin D1,NF-κB mRNA水平的表达,Western blot检测含药血清对PTEN,p-PI3K,p-AKT蛋白水平的表达。结果:低剂量补肾疏肝方含药血清对A549细胞有抑制作用,低剂量中药与顺铂联用有协同作用且以时间依赖方式,即补肾疏肝方低剂量组与联合顺铂低剂量组在48,72,96 h的抑制率均高于其他组(P<0.05,P<0.01);补肾疏肝方低剂量组与联合顺铂低剂量组拟态血管数目均低于其他组(P<0.05,P<0.01);补肾疏肝方低剂量组与联合顺铂低剂量组PTEN蛋白表达均高于其他组(P<0.05,P<0.01),补肾疏肝方低剂量组与联合顺铂低剂量组p-PI3K,p-AKT蛋白,Akt1,Cyclin D1,NF-κB的mRNA表达均低于其他组(P<0.05,P<0.01)。结论:补肾疏肝方含药血清可通过PTEN-PI3K-AKT信号通路抑制人肺腺癌A549细胞增殖和转移。Objective： To explore the inhibiting effect and mechanism of Bushen Shugan formula on A549 cells proliferation and metastasis by PTEN-PI3K-AKT pathway. Method： Sixty rats were randomly divided into 6groups： the normal saline group（2 m L）,the cisplatin group（8 mg·kg- 1）,the low-,high-dose Bushen Shugan formula groups（15,30 g·kg- 1）,and the low-dose Bushen Shugan formula plus cisplatin group（15 g·kg- 1＋ 8 mg·kg- 1）,the high-dose Bushen Shugan formula plus cisplatin group（30 g·kg- 1＋ 8 mg·kg- 1） of 10 rats each. The corresponding medicines were intragastrically administered to rats twice daily for 5 days and the drug serums were prepared. The cell proliferation ability of each group was evaluated by MTT. The capillary structure in each group was observed by three-dimensional cell culture and the number was counted. The mRNA expressions of AKT-1,Cyclin D1 and NF-κB were detected by using RT-PCR. The protein expressions of PTEN,p-PI3 K and p-AKT were detected by using Western blot. Result： Low-dose drug-contained serum of Bushen Shugan formula could inhibit the proliferation of A549 in a time-dependent manner. The inhibiting rates were lower after 48,72 and 96 h culturing,the numbers of tuber-shaped structure were lower,the protein expression of PTEN was higher,the protein expressions of p-PI3 K and p-AKT were lower,and the mRNA expressions of Akt1,Cyclin D1,NF-κB were lower in the Bushen Shugan formula plus cisplatin groups than the other group（P〈0. 05,P〈0. 01）. There were certain synergies between Bushen Shugan formula and cisplatin. Conclusion： Drug-contained serum of Bushen Shugan formula could inhibit the cell proliferation and metastasis of A549 cells by PTEN-PI3K-AKT signaling pathway.

关 键 词：补肾疏肝方 A549p PTEN-PI3K-AKT信号通路 

分 类 号：R285.5[医药卫生—中药学]