Nie Peipei, Hu Weicheng, Zhang Tao, Yang Yiju, Hou Benxin, Zou Zhengzhi
MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China.
Cell Physiol Biochem. 2015;35(6):2255-71. doi: 10.1159/000374030. Epub 2015 Apr 9.
BACKGROUND/AIM: Treatment of human non-small-cell lung cancer (NSCLC) often involves uses of multiple therapeutic strategies with different mechanisms of action. Here we found that resveratrol (RV) enhanced the anti-tumor effects of epidermal growth factor receptor (EGFR) inhibitor erlotinib in NSCLC cells.
Cell viability was measured by MTT assay and clonogenicity assay. Western blot was applied to assess the protein expression levels of target genes. Cell apoptosis was monitored by AnnexinV-FITC assay and sub-G1 population assay. Intracellular ROS were measured by flow cytometric analysis. Cell caspase activities were carried out by fluorometric assays.
Exposure of H460, A549, PC-9 and H1975 cells to minimal or non-toxic concentrations of RV and erlotinib synergistically reduced cell viability, colony formation and induced cell apoptosis. Furthermore, RV synergistically enhanced erlotinib-induced apoptosis was involved in ROS production. Additionally, co-treatment with RV and erlotinib repressed the expressions of anti-apoptosis proteins, such as survivin and Mcl-1, whereas promoted p53 and PUMA expression and caspase 3 activity. Moreover, the combination was also more effective at inhibiting the AKT/mTOR/S6 kinase pathway. Subsequently, small interfering RNA (siRNA) depletion of PUMA and overexpression of survivin significantly attenuated NSCLC cells apoptosis induced by the combination of the two drugs.
Our findings suggested that RV synergistically enhanced the anti-tumor effects of erlotinib in NSCLC cells were involved in decrease of survivin expression and induction of PUMA expression. In conclusion, based on the observations from our study, we indicated that the combined administration of these two drugs might be considered as a novel therapeutic regimen for treating NSCLC.
背景/目的：人类非小细胞肺癌（NSCLC）的治疗通常涉及多种作用机制不同的治疗策略。在此，我们发现白藜芦醇（RV）可增强表皮生长因子受体（EGFR）抑制剂厄洛替尼对NSCLC细胞的抗肿瘤作用。
采用MTT法和克隆形成试验检测细胞活力。应用蛋白质印迹法评估靶基因的蛋白表达水平。通过膜联蛋白V-异硫氰酸荧光素（AnnexinV-FITC）试验和亚G1期细胞群试验监测细胞凋亡。通过流式细胞术分析测量细胞内活性氧（ROS）。通过荧光测定法检测细胞半胱天冬酶活性。
将H460、A549、PC-9和H1975细胞暴露于最低或无毒浓度的RV和厄洛替尼下，可协同降低细胞活力、集落形成并诱导细胞凋亡。此外，RV协同增强厄洛替尼诱导的凋亡与ROS产生有关。此外，RV与厄洛替尼联合处理可抑制抗凋亡蛋白如生存素和髓细胞白血病-1（Mcl-1）的表达，而促进p53和p53上调凋亡调节因子（PUMA）的表达及半胱天冬酶3活性。此外，该联合用药在抑制AKT/雷帕霉素靶蛋白（mTOR）/S6激酶途径方面也更有效。随后，小干扰RNA（siRNA）敲低PUMA以及生存素过表达显著减弱了两种药物联合诱导的NSCLC细胞凋亡。
我们的研究结果表明，RV协同增强厄洛替尼对NSCLC细胞的抗肿瘤作用与生存素表达降低和PUMA表达诱导有关。总之，基于我们研究的观察结果，我们指出这两种药物联合给药可能被视为一种治疗NSCLC的新型治疗方案。
