Laboratory of Molecular Diagnosis of Cancer, West China Hospital of Sichuan University, Chengdu, China.
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Clin Exp Pharmacol Physiol. 2020 Jun;47(6):1083-1091. doi: 10.1111/1440-1681.13287. Epub 2020 Mar 20.
Carboplatin (CBP) is a widely used targeted anticancer therapeutic drug; however, multi-drug resistance induced by the accumulation of CBP eventually causes diseases progression. The anti-malarial drug artesunate (ART) also exerts anticancer effects in various cancers; however, the combined effect of ART and CBP on non-small cell lung cancer (NSCLC) remains unclear. In the present study, the NSCLC cell line A549 was pretreated with various concentrations of CBP, ART and gemcitabine (GEM). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were conducted to detect cell viability. Cell apoptosis was evaluated by both flow cytometry and TUNEL apoptotic assay. The expression profiles of cell cycle-related proteins and apoptotic proteins were determined by western blot. Cell clone numbers were visualized using crystal violet staining. Here, we found that both CBP and ART suppressed cell viability, and promoted cell apoptosis, and the combined application of ART and CBP at a lower concentration exhibited synergistic effects. Specifically, the combination of ART and CBP at a lower concentration suppressed cell clone numbers, promoted cell cycle arrest at the G /M phase, and induced the expression of the cell cycle and apoptosis-related proteins BAX, p21, p53, and Caspase-3, while decreasing Bcl-2 and Cyclin B1 expression. Based on these results, we concluded that combined application of ART and CBP exerts synergistic anti-tumour effects on NSCLC by enhancing cell apoptosis in a mitochondria-dependent manner.
卡铂(CBP)是一种广泛应用的靶向抗癌治疗药物;然而,CBP 积累引起的多药耐药最终导致疾病进展。抗疟药青蒿琥酯(ART)在各种癌症中也具有抗癌作用;然而,ART 和 CBP 联合用于非小细胞肺癌(NSCLC)的效果尚不清楚。在本研究中,NSCLC 细胞系 A549 先用不同浓度的 CBP、ART 和吉西他滨(GEM)预处理。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法检测细胞活力。通过流式细胞术和 TUNEL 凋亡测定法评估细胞凋亡。通过 Western blot 测定细胞周期相关蛋白和凋亡蛋白的表达谱。通过结晶紫染色观察细胞克隆数。在这里,我们发现 CBP 和 ART 均抑制细胞活力,促进细胞凋亡,ART 和 CBP 的联合应用在较低浓度下表现出协同作用。具体而言,ART 和 CBP 的联合应用在较低浓度下抑制细胞克隆数,促进细胞周期停滞在 G/M 期,并诱导细胞周期和凋亡相关蛋白 BAX、p21、p53 和 Caspase-3 的表达,同时降低 Bcl-2 和 Cyclin B1 的表达。基于这些结果,我们得出结论,ART 和 CBP 的联合应用通过以线粒体依赖性方式增强细胞凋亡对 NSCLC 发挥协同抗肿瘤作用。