| mTOR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 |
| 投稿时间：2017-12-25 修订日期：2018-03-09 点此下载全文 |
| 引用本文：丁木莲,李峰.mTOR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[J].医学研究杂志,2018,47(11):189-193 |
| DOI：
10.11969/j.issn.1673-548X.2018.11.042 |
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| 中文摘要:目的 探讨哺乳动物雷帕霉素靶蛋白（mTOR）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（IC50），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IC50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 mTOR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。 |
| 中文关键词:哺乳动物雷帕霉素靶蛋白 AZD8055 乳腺癌 多西紫杉醇 |
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| Effects of mTOR Highly Selective Inhibitor of AZD8055 on Docetaxel-Resistant Breast Cancer Cell Lines |
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| Abstract: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-7 cell lines and MCF-7/Doc cell lines were cultured in vitro.The median inhibitory concentration (IC50) of MCF-7/Doc cells on docetaxel in was detected by MTT assay. The experiment was divided into 2 groups, including MCF-7/Doc 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 AZD8055 was detected by tumor bearing 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-7 and MCF-7/Doc 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 IC50 of MCF-7/Doc+AZD8055 group (5.1±0.6 μmol/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 MDR1 gene 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). AZD8055 has significant inhibitory effect on transplanted tumor. The inhibition rate was 48.95%. The difference was statistically significant (P<0.05). Similar results were obtained by repeated experiments of MDA-MB-231 and MDA-MB-231/Doc cell lines. Conclusion mTOR highly selective inhibitor of AZD8055 can significantly increase the sensitivity of breast cancer cell lines to docetaxel. Its mechanism may be related to inhibiting the expression of MDR1 and P-gp. Further research is needed to confirm. |
| keywords:Mammalian target of rapamycin AZD8055 Breast cancer Docetaxel |
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