The First Clinical Medical College of Lanzhou University, Lanzhou, Gansu, China.
Key Laboratory of Gynecological Oncology of Gansu Province, Lanzhou, Gansu, China.
Bioengineered. 2022 Mar;13(3):7277-7292. doi: 10.1080/21655979.2022.2047496.
This study aimed to elucidate the specific anticancer mechanism of 6-methoxyflavone in HeLa cells. A total of 178 putative targets of 6-methoxyflavone were obtained from the PharmMapper database. Microarray analyses, transcriptome sequencing analyses, functional enrichment analyses, and gene set enrichment analyses were performed to preliminarily explore the roles and mechanisms of the 178 targets in cervical cancer. Cell counting kit-8, cell cycle assays, polymerase chain reactions, and western blotting were used to clarify the mechanism of action of 6-methoxyflavone. Molecular docking and noncovalent interaction analyses were performed to further confirm the mechanism of action in three-dimensional structures. Functional enrichment analyses and gene set enrichment analyses indicated that high mRNA expression of cyclin A2 (CCNA2) and cyclin-dependent kinase 2 (CDK2) stimulated cell cycle progression in cervical cancer. Cell proliferation and cycle assays, transcriptome sequencing, polymerase chain reactions, and western blotting revealed that 6-methoxyflavone inhibited HeLa cell proliferation and induced S-phase arrest via the CCNA2/CDK2/ cyclin-dependent kinase inhibitor 1A (p21CIP1) pathway. Molecular docking and noncovalent interaction analyses showed that 6-methoxyflavone had the strongest affinity toward, inhibitory effect on, and noncovalent interactions with CDK2, and that the combination of CDK2 and CCNA2 enhanced these effects. An analysis of clinical characteristics showed that 6-methoxyflavone might be related to six clinicopathological parameters of cervical cancer patients. 6-Methoxyflavone induces S-phase arrest in HeLa cells via the CCNA2/CDK2/p21CIP1 pathway.
本研究旨在阐明 6-甲氧基黄酮在 HeLa 细胞中的特定抗癌机制。从 PharmMapper 数据库中获得了 6-甲氧基黄酮的 178 个潜在靶标。进行了微阵列分析、转录组测序分析、功能富集分析和基因集富集分析，初步探讨了 178 个靶标在宫颈癌中的作用和机制。使用细胞计数试剂盒-8、细胞周期测定、聚合酶链反应和蛋白质印迹法阐明 6-甲氧基黄酮的作用机制。进行分子对接和非共价相互作用分析，以进一步确认三维结构中的作用机制。功能富集分析和基因集富集分析表明，cyclin A2（CCNA2）和细胞周期蛋白依赖性激酶 2（CDK2）的高 mRNA 表达促进宫颈癌中的细胞周期进程。细胞增殖和周期测定、转录组测序、聚合酶链反应和蛋白质印迹法显示，6-甲氧基黄酮通过 CCNA2/CDK2/细胞周期蛋白依赖性激酶抑制剂 1A（p21CIP1）途径抑制 HeLa 细胞增殖并诱导 S 期停滞。分子对接和非共价相互作用分析表明，6-甲氧基黄酮与 CDK2 具有最强的亲和力、抑制作用和非共价相互作用，并且 CDK2 与 CCNA2 的结合增强了这些作用。临床特征分析表明，6-甲氧基黄酮可能与宫颈癌患者的六个临床病理参数有关。6-甲氧基黄酮通过 CCNA2/CDK2/p21CIP1 途径诱导 HeLa 细胞 S 期停滞。
