The First Laboratory of Cancer Institute, the First Hospital of China Medical University, Shenyang, China.
Department of Oncology, the Second Affiliated Hospital of Dalian Medical University, Dalian, China.
FASEB J. 2021 May;35(5):e21601. doi: 10.1096/fj.202002780R.
Peritoneal dissemination threatens the survival of patients with gastric cancer (GC). Bufalin is an extract of traditional Chinese medicine, which has been proved to have anticancer effect. The target of bufalin in suppressing gastric cancer peritoneal dissemination (GCPD) and the underlying mechanism are still unclear. In this research, GC cell line MGC-803 and high-potential peritoneal dissemination cell line MKN-45P were treated with bufalin or L-NAME. Malignant biological behavior and protein level of GC cell lines were detected with MTT, wound healing, transwell, adhesion, and western blotting. Bioinformatics analysis and patient tissues were used to verify the role of endothelial nitric oxide synthase (NOS3) in GC. Mice model was used to assess the effect of bufalin and role of NOS3 in vivo. We found that bufalin inhibited the proliferation, invasion, and migration in GC cell lines. NOS3, which was an independent prognostic factor of GC patients, was predicted to be a potential target of bufalin. Further experiments proved that bufalin reduced the phosphorylation of NOS3, thereby inhibiting the mitogen-activated protein kinase (MAPK) signaling pathway, and ultimately suppressed GCPD by inhibiting EMT process. In conclusion, NOS3 was a potential therapeutic target and prognostic biomarker of GC. Bufalin could suppress GCPD through NOS3-MAPK signaling pathway, which provided more evidence support for intraperitoneal perfusion of bufalin to treat GCPD.
腹膜转移威胁着胃癌（GC）患者的生存。蟾毒灵是一种中药提取物，已被证明具有抗癌作用。蟾毒灵抑制胃癌腹膜转移（GCPD）的靶点及其潜在机制尚不清楚。在这项研究中，用蟾毒灵或 L-NAME 处理 GC 细胞系 MGC-803 和高潜能腹膜转移细胞系 MKN-45P。用 MTT、划痕愈合、transwell、黏附实验和 Western blot 检测 GC 细胞系的恶性生物学行为和蛋白水平。利用生物信息学分析和患者组织验证内皮型一氧化氮合酶（NOS3）在 GC 中的作用。用小鼠模型评估蟾毒灵的作用及其在体内对 NOS3 的作用。我们发现蟾毒灵抑制 GC 细胞系的增殖、侵袭和迁移。NOS3 是 GC 患者的独立预后因素，被预测为蟾毒灵的潜在靶点。进一步的实验证明，蟾毒灵降低了 NOS3 的磷酸化水平，从而抑制丝裂原活化蛋白激酶（MAPK）信号通路，最终通过抑制 EMT 过程抑制 GCPD。总之，NOS3 是 GC 的潜在治疗靶点和预后生物标志物。蟾毒灵可以通过 NOS3-MAPK 信号通路抑制 GCPD，为腹腔内灌注蟾毒灵治疗 GCPD 提供了更多的证据支持。
