苦参碱诱导人乳腺癌Bcap-37细胞发生自噬与自噬相关蛋白mTOR相关性研究  

Relationship Between Matrine Induced Autophagy and mTOR in Human Breast Cancer Bcap-37 Cells

作　　者：任莉莉[1] 王霖玲 王晓稼[3] 

机构地区：[1]浙江省肿瘤医院中西医结合科,杭州310022 [2]浙江中医药大学,杭州310053 [3]浙江省肿瘤医院肿瘤内科,杭州310022

出　　处：《浙江中西医结合杂志》2016年第9期783-786,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省医药卫生科技计划项目(No.2013KYB04)

摘　　要：目的探讨苦参碱是否可通过抑制mTOR活性阻断PI3K/Akt信号通路,诱导人乳腺癌Bcap-37细胞自噬发生。方法苦参碱(终浓度为0、1、2mg/m L)±氯喹(chloroquine)(自噬抑制剂)处理乳腺癌Bcap-37细胞24h,运用免疫印迹(Western-blot)法检测PI3K/Akt信号通路自噬相关蛋白mTOR、p-mTOR、p70S6K、p-p70S6、e IF4E表达。苦参碱(终浓度为0、1、2mg/m L)±Bafilomycin A1(自噬抑制剂)/Rapamcin(自噬诱导剂)处理乳腺癌Bcap-37细胞24h,Western-blot法检测e IF4E蛋白的表达。结果苦参碱剂量依赖性抑制mTOR、p70S6k磷酸化,同时降低e IF4E蛋白表达;当苦参碱联合氯喹(CQ)作用于乳腺癌Bcap-37细胞时,则对上述蛋白表达均无明显抑制作用。苦参碱联合Bafilomycin A1作用于Bcap-37细胞时,e IF4E蛋白表达同样无明显改变;苦参碱联合Rapamcin时,e IF4E蛋白表达则呈剂量依赖性降低。结论苦参碱可通过抑制mTOR活性阻断PI3K/Akt信号通路,从而诱导人乳腺癌Bcap-37细胞自噬发生。Objective To investigate whether the matrine induce autophagy in human breast cancer Bcap-37 cells by down-regulating PI3K/Akt signal pathway via inhibition of mTOR or not. Methods Human breast cancer Bcap-37 cells were cultured in vitro and treated by different final concentrations （0, 1, 2 mg/mL） of matrine ± chloroquine（autophagy inhibitor）, then cultured for 24 h. Western-blot assay was used to detect the protein expres-sion level of mTOR, p-mTOR, p70S6K, p-p70S6, eIF4E, which were related to PI3K/Akt signaling pathway. The same cells were cultured in vitro and treated by matrine （0, 1, 2 mg/mL） ± bafilomycinA1 （autophagy inhibitor）/rapamcin （autophagy revulsant） for 24 h. Western-blot assay was used to detect the level of eIF4E. Results The expression of p-mTOR, p-p70S6k and eIF4E proteins down-regulated after Bcap-37 cells were treated with matrine at 0, 1, 2 mg/mL for 24 h in a does-dependent manner, but had no obvious changes after Bcap-37 cells were treated with matrine ＋ chloroquine for the same while. The expression of eIF4E protein also had no obvious change after Bcap-37 cells were treated with matrine ＋ bafilomycinA1 for 24 h, however the expression of eIF4E protein down-regulated after the cells were treated with matrine＋ rapamcin in a does dependent manner. Conclu-sion The molecular mechanism of autophagy induced by the effect of matrine in human breast cancer Bcap-37 cells may be via down-regulating PI3K/Akt/mTOR signaling pathway.

关 键 词：苦参碱 Bcap-37细胞 自噬 MTOR P70S6K EIF4E 

分 类 号：R285[医药卫生—中药学]