摘要
目的探讨哺乳动物雷帕霉素靶蛋白(m TOR)高选择性抑制剂AZD8055对乳腺癌多西紫杉醇耐药细胞株化疗敏感度的作用。方法体外培养MCF-7细胞株、MCF-7/Doc细胞株。噻唑蓝比色法检测AZD8055作用后MCF-7/Doc细胞对多西紫杉醇的半数抑制浓度(IC_(50)),实验分2组:MCF-7/Doc组和MCF-7/Doc+AZD8055组。实时荧光定量PCR和蛋白印迹法检测AZD8055作用后,细胞MDR1基因和P-gp蛋白的表达,实验分4组:MCF-7组、MCF-7+AZD8055组、MCF-7/Doc组、MCF-7/Doc+AZD8055组。裸鼠体内荷瘤实验检测AZD8055的体内抑瘤率,实验分3组:空白对照组、阴性对照组和AZD8055组。为排除MCF-7和MCF-7/Doc细胞特异性对实验结果的影响,研究采用乳腺癌MDA-MB-231和MDA-MB-231/Doc细胞株重复了上述实验。结果噻唑蓝比色法检测显示,MCF-7/Doc+AZD8055组细胞对多西紫杉醇的IC_(50)为5. 1±0. 6μmol/L,明显低于MCF-7/Doc组(P <0. 05),AZD8055对多西紫杉醇的耐药逆转倍速为5. 4。实时荧光定量PCR检测显示,MCF-7/Doc组细胞MDR1基因表达量为1. 84±0. 35,明显高于MCF-7组(P <0. 05);而MCF-7/Doc+AZD8055组细胞MDR1基因表达量为1. 21±0. 29,明显低于MCF-7/Doc组(P <0. 05)。蛋白印迹法检测显示,MCF-7/Doc组细胞P-gp蛋白表达量分别为0. 93±0. 15,明显高于MCF-7组(P <0. 05);而MCF-7/Doc+AZD8055组细胞P-gp蛋白表达量0. 54±0. 10,明显低于MCF-7/Doc组(P <0. 05)。AZD8055组肿瘤生长速度明显低于空白对照组和阴性对照组(P <0. 05),AZD8055对移植瘤具有显著的抑制作用,抑瘤率为48. 95%,差异有统计学意义(P <0. 05)。MDA-MB-231和MDA-MB-231/Doc细胞株重复实验得到了相似结果。结论 m TOR高选择性抑制剂AZD8055可显著提高乳腺癌细胞株对多西紫杉醇的化疗敏感度,其作用机制可能与抑制MDR1和P-gp表达有关。
Objective To investigate the effects of mammalian target of rapamycin (mTOR)highly selective inhibitor of AZD8055 on docetaxel -resistant breast cancer cell lines.Methods MCF -7cell lines and MCF -7/Doc cell lines were cultured in vitro.The median inhibitory concentration (IC50)of MCF -7/Doe cells on docetaxel in was detected by MTT assay.The experiment was divided into 2 groups,including MCF -7/Doe group and MCF -7/Doc +AZD8055 group.The expressions of MDR1 gene and P -gp protein were detected by real-time quantitative PCR and Western blot.The experiment was divided into 4 groups,including MCF -7 group,MCF -7+ AZD8055 group,MCF-7/Doc group and MCF-7/Doc +AZD8055 group.The tumor inhibitory rate of AZD8055was detected by tumor hearing test.The experiment was divided into 3 groups:blank control group,negative control group and AZD8055 group.In order to exclude the effects of MCF -7and MCF-7/Doe cell specificity on the experimental results,this study repeated these experiments using MDA -MB -231 and MDA -MB -231/Doc cell lines of breast cancer.Results MTT assay showed that the IC50of MCF-7/Doc + AZDS055 group (5.1±0.6μmol/L)was significantly lower than the IC50 in MCF -7/Doc group (P <0.05),and the reveral fold of AZD8055 was 5.4.Real-time quantitative PCR showed that the expression of MDR1gene in MCF -7/Doc group (1.84±0.35)was significantly higher than that in MCF-7 group (P <0.05),and the expression of MDR1 gene in MCF-7/Doc +AZD8055 group (1.21±0.29)was significantly lower than that in MCF-7/Doc group (P <0.05).Western blot showed that the expression of P -gp protein in MCF -7/Doc group (0.93±0.15)was significantly higher than that in MCF-7 group (P <0.05),and the expression of P-gp protein in MCF -7/Doc+AZD8055 group (0.54±0.10)was significantly lower than that in MCF -7/Doc group (P <0.05).The tumor inhibitory rate in AZD8055 group was significantly lower than the blank control group and negative control group (P <0.05).AZD8055has significant inhibitory effect on transplanted tumor.The inhibition rate wa
作者
丁木莲
李峰
Ding Mulian;Li Feng(Pharmacy of Pengzhou People's Hospital,Sichuan 611930,China)
出处
《医学研究杂志》
2018年第11期189-193,共5页
Journal of Medical Research
