类黄酮非瑟酮对人头颈鳞状细胞癌 (HNSCC) 的抗癌潜力和相关机制尚未得到充分研究。在本研究中，非瑟酮（25-75μM，24-48小时）剂量依赖性地抑制HNSCC Cal33和UM-SCC-22B细胞的生长并诱导死亡，而在正常细胞中没有显示任何死亡。 Fisetin (25-50µM) 通过减少 Cdc25C、CDK1、细胞周期蛋白 B1 表达和增加 p53(S15) 诱导 G2/M 期停滞。通过彗星测定、γ-H2A.X(S139) 磷酸化和 PARP 蛋白的显着裂解，证实了 HNSCC 细胞中非瑟酮诱导的 DNA 损伤和细胞凋亡的浓度依赖性增加。有趣的是，非瑟酮诱导的细胞死亡独立于 p53 和活性氧的产生而发生。确定了 JNK 的激活和 PI3K/Akt、ERK1/2、EGFR 和 STAT-3 信号传导的抑制。此外，非瑟酮诱导的细胞凋亡部分是通过 caspase 裂解 p21Cip1 和 p27Kip1 介导的，而 z-VAD-FMK（一种泛 caspase 抑制剂）可逆转这一过程。随后，非瑟酮也被发现可以诱导自噬；然而，自噬减弱会加剧细胞凋亡。口服非瑟酮（50mg/kg体重）治疗小鼠Cal33异种移植物19天，肿瘤体积抑制73%（p<0.01），同时Ki67阳性细胞减少，裂解的caspase-3水平增加在肿瘤中。与 50 µM 非瑟汀的体外作用一致，非瑟汀也降低了肿瘤中 p21Cip1 和 P27Kip1 的蛋白水平。总之，这些研究结果表明，非瑟酮对 HNSCC 具有强大的抗癌功效，可下调 EGFR-Akt/ERK1/2-STAT-3 通路并激活 JNK/c-Jun、半胱天冬酶以及半胱天冬酶介导的 p21Cip1 和 p27Kip1 裂解。© 2024 Wiley期刊有限责任公司。

The anticancer potential and associated mechanisms of flavonoid fisetin are yet to be fully investigated on human head and neck squamous cell carcinoma (HNSCC). In the present study, fisetin (25-75 µM for 24-48 h) dose-dependently inhibited growth and induced death in HNSCC Cal33 and UM-SCC-22B cells, without showing any death in normal cells. Fisetin (25-50 µM) induced G2/M phase arrest via decrease in Cdc25C, CDK1, cyclin B1 expression, and an increase in p53(S15). A concentration-dependent increase in fisetin-induced DNA damage and apoptosis in HNSCC cells was authenticated by comet assay, gamma-H2A.X(S139) phosphorylation, and marked cleavage of PARP protein. Interestingly, fisetin-induced cell death occurred independently of p53 and reactive oxygen species production. The activation of JNK and inhibition of PI3K/Akt, ERK1/2, EGFR, and STAT-3 signaling were identified. Further, fisetin-induced apoptosis was mediated, in part, via p21Cip1 and p27Kip1 cleavage by caspase, which was reversed by z-VAD-FMK, a pan-caspase inhibitor. Subsequently, fisetin was also found to induce autophagy; nevertheless, autophagy attenuation exaggerated apoptosis. Oral fisetin (50 mg/kg body weight) treatment to establish Cal33 xenograft in mice for 19 days showed 73% inhibition in tumor volume (p < 0.01) along with a decrease in Ki67-positive cells and an increase in cleaved caspase-3 level in tumors. Consistent with the effect of 50 µM fisetin in vitro, the protein levels of p21Cip1 and P27Kip1 were also decreased by fisetin in tumors. Together, these findings showed strong anticancer efficacy of fisetin against HNSCC with downregulation of EGFR-Akt/ERK1/2-STAT-3 pathway and activation of JNK/c-Jun, caspases and caspase-mediated cleavage of p21Cip1 and p27Kip1.© 2024 Wiley Periodicals LLC.