Department of Cell Biology, College of Integrated Chinese and Western Medicine, Liaoning University of Traditional Chinese Medicine, 79 Chongshan Eastern Road, Huanggu District, Shenyang 110847, China.
Key Laboratory of Ministry of Education for Traditional Chinese Medicine (TCM) Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, 79 Chongshan Eastern Road, Huanggu District, Shenyang 110847, China.
Biomed Res Int. 2022 Sep 1;2022:6022981. doi: 10.1155/2022/6022981. eCollection 2022.
Cisplatin resistance is one of the major obstacles in the treatment of nonsmall cell lung cancer (NSCLC). Kangai injection (KAI), a Chinese herbal medicine, has been used in tumors as adjuvant treatment, but its exact antitumor mechanism is still unclear. In this study, we first demonstrated that cisplatin-resistant A549/DDP cells showed a higher level of basal autophagy in response to cisplatin treatment with increasing autophagic protein expression levels of Beclin 1, p62, and LC3 compared to cisplatin-sensitive A549/DDP cells; then, we assessed the antitumor effect of KAI in cisplatin-resistant lung adenocarcinoma A549/DDP cells. Our results showed that KAI exhibited direct cytotoxic and chemosensitizing effects in A549/DDP cells. Combining KAI with cisplatin promoted A549/DDP cell apoptosis, which was confirmed by cell cycle arrest, condensed nuclear chromatin, annexin V fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) staining, and apoptosis-related protein expression. In addition, combining KAI with cisplatin induced autophagic cell death in A549/DDP cells with a high level of basal autophagy, as indicated by an increase in LC3 spot count, an accumulation of Beclin 1 and LC3 II, and reduced p62 protein expression. We also found that the apoptosis and autophagic cell death induced by cotreatment of KAI and cisplatin in A549/DDP cells were FOXO3a-dependent as indicated by decreased p-FOXO3a expression and increased FOXO3a nuclear localization, respectively. Furthermore, the gene knockdown assay further confirmed that KAI enhanced cisplatin cytotoxicity in A549/DDP cells with a high level of basal autophagy by inducing apoptosis and autophagic cell death in a FOXO3a-dependent manner. These findings suggest that the combination of KAI and cisplatin might support the potential clinical treatment as a novel strategy to overcome cisplatin resistance.
顺铂耐药是治疗非小细胞肺癌（NSCLC）的主要障碍之一。康艾注射液（KAI）是一种中药，已被用于肿瘤的辅助治疗，但确切的抗肿瘤机制仍不清楚。在这项研究中，我们首先证明顺铂耐药 A549/DDP 细胞在顺铂处理下表现出更高水平的基础自噬，与顺铂敏感 A549/DDP 细胞相比，自噬蛋白 Beclin 1、p62 和 LC3 的表达水平增加；然后，我们评估了 KAI 在顺铂耐药肺腺癌 A549/DDP 细胞中的抗肿瘤作用。结果表明，KAI 对 A549/DDP 细胞具有直接的细胞毒性和化疗增敏作用。将 KAI 与顺铂联合使用可促进 A549/DDP 细胞凋亡，这通过细胞周期阻滞、浓缩核染色质、Annexin V-FITC/PI 染色和凋亡相关蛋白表达得到证实。此外，在具有高基础自噬水平的 A549/DDP 细胞中，KAI 与顺铂联合使用诱导自噬性细胞死亡，表现为 LC3 斑点计数增加、Beclin 1 和 LC3 II 积累以及 p62 蛋白表达减少。我们还发现，KAI 和顺铂联合处理 A549/DDP 细胞诱导的凋亡和自噬性细胞死亡与 FOXO3a 依赖性有关，表现为 p-FOXO3a 表达减少和 FOXO3a 核定位增加。此外，基因敲低实验进一步证实，KAI 通过诱导 FOXO3a 依赖性凋亡和自噬性细胞死亡增强了 A549/DDP 细胞中高基础自噬水平的顺铂细胞毒性。这些发现表明，KAI 与顺铂联合可能支持作为克服顺铂耐药的新策略的潜在临床治疗。
