Hsieh Tsung-Han, Kuo Han-Pin, Chen Mei-Chuan, Lin Yu-Chen, Lin Bo-Jyun, Hsu Kai-Cheng, Chen Chun-Han
Precision Health Center, Taipei Medical University, Taipei, 110301, Taiwan.
Center of Thoracic Medicine, Taipei Medical University, Taipei, 110301, Taiwan.
Sci Rep. 2025 Jul 1;15(1):20922. doi: 10.1038/s41598-025-06543-3.
Bladder cancer (BC) remains a major therapeutic challenge, particularly in patients with acquired resistance to platinum-based chemotherapy. In this study, we investigated the potential of hinokiflavone (HNK), a natural biflavonoid, as a therapeutic agent against cisplatin-resistant BC. Our results demonstrate that HNK differentially inhibited the proliferation of cisplatin-resistant BC cells while sparing normal uroepithelial cells. Mechanistically, HNK induced apoptosis through both intrinsic and extrinsic pathways, as evidenced by caspase activation and Annexin V staining. Next-generation sequencing and gene set enrichment analysis revealed that HNK modulates genes involved in biosynthesis, metabolism, DNA replication and DNA repair. Additionally, HNK downregulated the transcription of MUTYH, OGG1, and XRCC1, which are key genes in base excision repair. For the first time, we identified that HNK as a novel inhibitor of CK2α via in vitro kinase assays, substrate phosphorylation assays, and molecular docking analysis. HNK treatment reduced the phosphorylation of known CK2α targets, including Akt, Stat3, and XRCC1, in cisplatin-resistant BC cells. Time-course analysis revealed that the inhibition of Akt phosphorylation coincided with PARP cleavage, and genetic rescue experiments confirmed the involvement of the CK2α/Akt axis in HNK-induced apoptosis. Furthermore, combination treatment with HNK and chemotherapeutic agents such as doxorubicin or mitomycin C resulted in enhanced cytotoxic effects, suggesting a potential role of HNK as a chemosensitizing agent. HNK, by targeting both DNA repair pathways and CK2-mediated survival signaling, may serve as a promising therapeutic candidate for cisplatin-resistant BC.
膀胱癌（BC）仍然是一个重大的治疗挑战，尤其是对于那些对铂类化疗产生获得性耐药的患者。在本研究中，我们调查了天然双黄酮扁柏黄酮（HNK）作为一种抗顺铂耐药性BC治疗药物的潜力。我们的结果表明，HNK能差异性地抑制顺铂耐药性BC细胞的增殖，同时对正常尿路上皮细胞无损害。从机制上来说，HNK通过内源性和外源性途径诱导细胞凋亡，这通过半胱天冬酶激活和膜联蛋白V染色得以证明。下一代测序和基因集富集分析显示，HNK调节参与生物合成、代谢、DNA复制和DNA修复的基因。此外，HNK下调了碱基切除修复中的关键基因MUTYH、OGG1和XRCC1的转录。通过体外激酶测定、底物磷酸化测定和分子对接分析，我们首次确定HNK是CK2α的新型抑制剂。HNK处理降低了顺铂耐药性BC细胞中已知的CK2α靶点（包括Akt、Stat3和XRCC1）的磷酸化水平。时间进程分析显示，Akt磷酸化的抑制与PARP裂解同时发生，基因拯救实验证实了CK2α/Akt轴参与了HNK诱导的细胞凋亡。此外，HNK与阿霉素或丝裂霉素C等化疗药物联合治疗可增强细胞毒性作用，表明HNK作为一种化学增敏剂具有潜在作用。通过靶向DNA修复途径和CK2介导的生存信号，HNK可能成为顺铂耐药性BC的一种有前景的治疗候选药物。
