2020 年第 10 期 第 15 卷

解毒祛瘀方通过抑制磷脂酰肌醇-3-激酶/蛋白激酶B信号通路逆转乳腺癌细胞他莫昔芬耐药的机制研究

Effect of Jiedu Quyu recipe on tamoxifen-resistance reversal of breast cancer cells by inhibiting phosphatidylinositol 3 kinase/protein kinase B pathway

作者：万斯斯1赵萍1姜华2黄际薇1

英文作者：Wan Sisi1 Zhao Ping1 Jiang Hua2 Huang Jiwei1

单位：1中山大学附属第三医院药剂科，广州510630；2中山大学附属第三医院甲状腺乳腺外科，广州510630

英文单位：1Department of Pharmacy the Third Affiliated Hospital of Sun Yat-Sen University Guangzhou 510630 China; 2Department of Nail and Breast Surgery the Third Affiliated Hospital of Sun Yat-Sen University Guangzhou 510630 China

关键词：乳腺癌；解毒祛瘀方；他莫昔芬耐药；磷脂酰肌醇-3-激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路

英文关键词：Breastcancer；JieduQuyurecipe；Tamoxifen-resistance；Phosphatidylinositol-3-kinases/protein-serine-threoninekinase/mammaliantargetofrapamycinsignalpathway

• 摘要：

• 目的 探讨解毒祛瘀方通过抑制磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（Akt）信号通路对乳腺癌细胞他莫昔芬耐药的逆转作用及机制。方法采用低浓度逐步加量诱导法建立乳腺癌他莫昔芬耐药细胞株MCF-7C，并分为空白对照组、解毒祛瘀方（2.5 g/L）组、他莫昔芬（10 μmol/L）组、他莫昔芬（10 μmol/L）+解毒祛瘀方（2.5 g/L）组、胰岛素样生长因子1（IGF-1）（10 μg/L）+他莫昔芬（10 μmol/L）+解毒祛瘀方（2.5 g/L）组。采用噻唑蓝法验证MCF-7C细胞的耐药性、解毒祛瘀方的细胞毒性以及对MCF-7C细胞他莫昔芬耐药性的影响。利用流式细胞术检测细胞凋亡率。利用蛋白质印迹法检测各组MCF-7C细胞中PI3K/Akt/雷帕霉素靶蛋白（mTOR）信号通路相关蛋白的表达。结果 MCF-7C细胞对他莫昔芬的耐药指数为7.20；而经解毒祛瘀方（2.5 g/L）协同作用后，MCF-7C细胞对他莫昔芬的耐药指数为2.02。他莫昔芬+解毒祛瘀方组MCF-7C细胞凋亡率明显高于他莫昔芬组［（56.72±7.25）%比（13.24±0.76）%］，同时p-PI3K［（0.37±0.06）比（0.68±0.07）］、p-Akt［（0.17±0.02）比（0.51±0.05）］、p-mTOR［（0.10±0.02）比（0.39±0.02）］蛋白表达量也明显低于他莫昔芬组（均P＜0.05）。而加入PI2K/Akt通路激活剂IGF-1后，IGF-1+他莫昔芬+解毒祛瘀方组MCF-7C细胞增殖抑制率和凋亡率较他莫昔芬+解毒祛瘀方组均明显降低（均P＜0.05）。结论 解毒祛瘀方可逆转乳腺癌细胞他莫昔芬的耐药性，其作用机制可能与抑制PI3K/Akt/mTOR信号通路活化有关。

• Objective To explore the reversal effect and mechanism of Jiedu Quyu recipe on tamoxifen resistance in breast cancer cells by inhibiting phosphatidylinositol 3 kinase (PI3K) / protein kinase B (Akt) signal pathway.Methods The tamoxifen-resistant MCF-7C was established by exposure of parental MCF-7 cells and increasing tamoxifen concentrations. MCF-7C cells were divided into control group, TAM (tamoxifen 10 μmol/L) group, TCM (traditional Chinese medicine, Jiedu Quyu recipe 2.5 g/L) group, TAM+TCM group and IGF-1(insulin growth factor-1, 10 μg/L)+TAM+TCM group. The tamoxifen-resistance of MCF-7C cells, cytotoxicity of Jiedu Quyu recipe and influence of Jiedu Quyu recipe on tamoxifen-resistance of MCF-7C cells were examined by Methyl thiazolyl tetrazolium (MTT). Flow cytometry was used to detect the apoptosis ratio. In addition, the expression of PI3K/Akt/mammalian target of rapamycin (mTOR) pathway related proteins were analyzed by western blot. Results The resistance index (RI) of MCF-7C cells on tamoxifen was 7.20. Under the synergism of Jiedu Quyu recipe, RI of MCF-7C cells on tamoxifen was 2.02. The apoptosis rate of MCF-7C cells in TAM+TCM group was higher than that in TAM group ［(56.72±7.25)% vs (13.24±0.76)%］(P＜0.05); the levels of p-PI3K ［(0.37±0.06) vs (0.68±0.07)］, p-Akt ［(0.17±0.02) vs (0.51±0.05)］, p-mTOR ［(0.10±0.02) vs (0.39±0.02)］ in TAM+TCM group were all lower than those in TAM group (all P＜0.05). The proliferation inhibition rate and apoptosis rate of MCF-7C cells in IGF-1+TAM+TCM group were lower than those in TAM+TCM group (both P＜0.05). Conclusion Jiedu Quyu recipe can reverse the resistance to tamoxifen of MCF-7C cells, which may be related with the inhibition of PI3K/Akt/mTOR signal pathway.