背景:肺癌,尤其是非小细胞肺癌(NSCLC),是全球癌症相关死亡的主要原因之一。长春新碱(VCR)是肺癌的化学治疗剂;然而,其有效性受到副作用和耐药性发展的限制。广藿香醇(PA),来自Pogostemoncablin提取物,已知具有抗炎和抗癌特性。在这项研究中,我们研究了PA在A549和VCR耐药的A549/V16NSCLC细胞中诱导活性氧(ROS)介导的DNA损伤中的作用。
方法:使用MTT法研究了PA的抗癌潜力,菌落形成,流式细胞术分析,西方印迹,DCFDA染色,免疫荧光染色,和TUNEL测定技术。
结果:PA处理的细胞内ROS水平增强,激活CHK1和CHK2信号通路。PA通过调节p53/p21和CDK2/cyclinE1的表达进一步抑制增殖和集落形成能力,并在G0/G1期诱导细胞周期停滞。此外,PA通过激活Bax/caspase-9/caspase-3内在途径增加处于亚G1期的细胞百分比并诱导细胞凋亡。此外,PA治疗后,耐药性(P-糖蛋白)和癌症干细胞(CD44和CD133)标志物下调。此外,PA和顺铂联合在A549和A549/V16细胞中表现出协同抑制活性。
结论:PA诱导ROS介导的DNA损伤,触发细胞周期停滞和凋亡,减毒耐药性和肿瘤干细胞表型,与顺铂联合使用可协同抑制增殖。这些发现表明PA具有用于治疗具有或不具有VCR抗性的NSCLC的潜力。
Lung cancer, especially non-small cell lung cancer (NSCLC), is one of the leading causes of cancer-related deaths worldwide. Vincristine (VCR) is a chemotherapeutic agent for lung cancers; however, its effectiveness is limited by side effects and the development of drug resistance. Patchouli alcohol (PA), from Pogostemon cablin extract, is known to possess anti-inflammatory and anticancer properties. In this study, we investigated the role of PA in inducing reactive oxygen species (ROS)-mediated DNA damage in A549 and VCR-resistant A549/V16 NSCLC cells.
The anticancer potential of PA was studied using the MTT assay, colony formation, flow cytometry analysis, western blotting, DCFDA staining, immunofluorescence staining, and TUNEL assay techniques.
The intracellular ROS levels were enhanced in PA-treated cells, activating the CHK1 and CHK2 signaling pathways. PA further inhibited proliferation and colony-forming abilities and induced cell cycle arrest at the G0 /G1 phase by regulating p53/p21 and CDK2/cyclin E1 expression. Moreover, PA increased the percentage of cells in the subG1 phase and induced apoptosis by activating the Bax/caspase-9/caspase-3 intrinsic pathway. In addition, drug resistance (p-glycoprotein) and cancer stem cell (CD44 and CD133) markers were downregulated after PA treatment. Furthermore, combining PA and cisplatin exhibited synergistic inhibitory activity in A549 and A549/V16 cells.
PA induced ROS-mediated DNA damage, triggered cell cycle arrest and apoptosis, attenuated drug resistance and cancer stem cell phenotypes, and synergistically inhibited proliferation in combination with cisplatin. These findings suggest that PA has the potential to be used for the treatment of NSCLC with or without VCR resistance.