论文部分内容阅读
目的:观察曲妥珠单抗(Trastuzumab)与转录信号转导子与激活子3蛋白(STAT3)抑制剂NSC 74859联用对曲妥珠耐药细胞株SK-BR-3R的生长抑制作用及机理研究。方法:采用四甲基偶氮唑蓝(MTT)法鉴定曲妥珠耐药的SK-BR-3R细胞株并检测曲妥珠单药处理、NSC 74859单药处理以及两药联用处理对SK-BR-3R细胞的生长抑制程度。建立SK-BR-3R的皮下肿瘤模型,观察两药联用对肿瘤生长的抑制效果;通过免疫印迹(Western Blot)实验检测SK-BR-3R细胞中磷酸化HER2(p-HER2),磷酸化STAT3(p-STAT3)及磷酸化AKT(p-AKT)的水平。结果:当曲妥珠浓度在50 nmol/L及NSC 74859的浓度在50μmol/L联用时,较之两药单用显示了显著的抑制效果,其差异具有统计学意义;进一步在建立的SK-BR-3R小鼠肿瘤模型中观察到了曲妥珠联合NSC74859治疗组显示了比曲妥珠或NSC 74859单独使用时更显著的抑瘤效果。最后,免疫印迹实验显示了曲妥珠和NSC74859联合处理显著降低了SK-BR-3R细胞的HER2,STAT3及AKT的磷酸化水平。结论:曲妥珠单抗联合NSC 74859使用可显著抑制曲妥珠耐药的乳腺癌细胞SK-BR-3R的生长,其机制可能是药物协同抑制了对肿瘤生长重要的PI3K/AKT信号通路。本研究可为临床上治疗曲妥珠耐药的乳腺癌提供参考。
OBJECTIVE: To observe the growth inhibitory effect and mechanism of trastuzumab combined with transcriptional signal transducer and activator of transcription 3 (STAT3) inhibitor NSC 74859 on trastuzumab-resistant cell line SK-BR-3R the study. METHODS: Trastuzumab-resistant SK-BR-3R cell line was identified by MTT assay and the effects of trastuzumab alone, NSC 74859 single-agent treatment, The extent of growth inhibition of -BR-3R cells. A subcutaneous tumor model of SK-BR-3R was established to observe the inhibitory effect of the two drugs on tumor growth. The phosphorylation of HER2 (p-HER2), phosphorylation and phosphorylation of SK-BR-3R cells were detected by Western Blot STAT3 (p-STAT3) and phosphorylated AKT (p-AKT). Results: When the concentration of trastuzumab was 50 nmol / L and the concentration of NSC 74859 was 50 μmol / L, the difference was statistically significant when compared with the two drugs alone. The difference was statistically significant between the established SK- It was observed in the BR-3R mouse tumor model that trastuzumab in combination with the NSC74859 treatment group showed more pronounced antitumor efficacy than either trastuzumab or NSC 74859 alone. Finally, Western blotting experiments showed that the combination of trastuzumab and NSC74859 significantly reduced the phosphorylation of HER2, STAT3 and AKT in SK-BR-3R cells. Conclusion: Trastuzumab combined with NSC 74859 can significantly inhibit the growth of trastuzumab-resistant breast cancer cell line SK-BR-3R. The mechanism may be that the drug synergistically inhibits the PI3K / AKT signaling pathway that is important for tumor growth. This study may provide a reference for the clinical treatment of trastuzumab-resistant breast cancer.