Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Jilin University, Changchun, Jilin 130021, P.R. China.
Department of Cardiovascular Medicine, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.
Mol Med Rep. 2021 Feb;23(2). doi: 10.3892/mmr.2020.11797. Epub 2020 Dec 23.
Neochlorogenic acid (NCA), a natural compound found in honeysuckle, possesses prominent anti‑inflammatory and antitumor effects. Pingyangmycin (PYM) induces DNA damage and has been used for the treatment of oral and maxillofacial tumors. Oral care serves an important role in promoting wound healing during chemotherapy in patients with oral squamous cell carcinoma (OSCC). Therefore, the present study aimed to analyze the effects of NCA and PYM on OSCC cells and to investigate the potential underlying mechanism. Reverse transcription‑quantitative PCR and western blotting were conducted to analyze the expression levels of DNA topoisomerase II α (TOP2A) in different OSCC cell lines. TOP2A‑overexpression cells were constructed via transfection of TOP2A‑overexpression plasmids. Following NCA or PYM treatment, cell proliferation was assessed using Cell Counting Kit‑8 and colony formation assays, whereas cell apoptosis and the cell cycle distribution were assessed via TUNEL staining and flow cytometry, respectively. In addition, the expression levels of apoptosis‑ and cell cycle‑related proteins were detected via western blotting. Moreover, co‑immunoprecipitation (Co‑IP) was conducted to determine whether TOP2A interacted with CDK1. The results of the present study indicated that NCA treatment significantly enhanced the suppressive effects of PYM on OSCC cell proliferation and apoptosis. The results also indicated that PYM arrested the cell cycle in the G/ by regulating cyclin dependent kinase 1 (CDK1)/cyclin B1, which was enhanced by the cotreatment of NCA and PYM. In addition, NCA and PYA treatment altered the expression levels of apoptosis‑related proteins. The Co‑IP assay indicated that TOP2A interacted with CDK1. Moreover, TOP2A overexpression significantly reversed the effects of NCA and PYM treatment on OSCC cell proliferation and apoptosis. In addition, NCA significantly decreased PYM‑induced toxicity in normal oral epithelial cells. In conclusion, the results of the present study suggested that NCA may promote the inhibitory effects of PYM in OSCC via TOP2A.
新绿原酸（NCA）是金银花中的一种天然化合物，具有显著的抗炎和抗肿瘤作用。平阳霉素（PYM）诱导 DNA 损伤，已用于治疗口腔颌面部肿瘤。口腔护理在促进口腔鳞状细胞癌（OSCC）患者化疗期间伤口愈合方面起着重要作用。因此，本研究旨在分析 NCA 和 PYM 对 OSCC 细胞的影响，并探讨其潜在的作用机制。通过反转录定量 PCR 和 Western blot 分析不同 OSCC 细胞系中 DNA 拓扑异构酶 IIα（TOP2A）的表达水平。通过转染 TOP2A 过表达质粒构建 TOP2A 过表达细胞。用细胞计数试剂盒-8 和集落形成实验评估 NCA 或 PYM 处理后细胞增殖，用 TUNEL 染色和流式细胞术分别评估细胞凋亡和细胞周期分布。此外，通过 Western blot 检测凋亡和细胞周期相关蛋白的表达水平。此外，还进行了共免疫沉淀（Co-IP）以确定 TOP2A 是否与 CDK1 相互作用。本研究结果表明，NCA 处理显著增强了 PYM 对 OSCC 细胞增殖和凋亡的抑制作用。结果还表明，PYM 通过调节细胞周期蛋白依赖性激酶 1（CDK1）/细胞周期蛋白 B1 将细胞周期阻滞在 G1 期，这一作用在 NCA 和 PYM 联合治疗时得到增强。此外，NCA 和 PYA 处理改变了凋亡相关蛋白的表达水平。Co-IP 实验表明 TOP2A 与 CDK1 相互作用。此外，TOP2A 过表达显著逆转了 NCA 和 PYM 处理对 OSCC 细胞增殖和凋亡的影响。此外，NCA 显著降低了 PYM 对正常口腔上皮细胞的毒性。综上所述，本研究结果表明，NCA 可能通过 TOP2A 促进 PYM 对 OSCC 的抑制作用。
