Key Laboratory of Radiation Oncology of Taizhou, Radiation Oncology Institute of Enze Medical Health Academy, Department of Radiation Oncology, Affiliated Taizhou Hospital of Wenzhou Medical University, Taizhou, 317000 Zhejiang Province, China.
Department of Pulmonary Medicine, At Enze Hospital, Affiliated Taizhou Hospital of Wenzhou Medical University, Taizhou, 317000 Zhejiang Province, China.
Anal Cell Pathol (Amst). 2022 Jun 16;2022:4484211. doi: 10.1155/2022/4484211. eCollection 2022.
Small-molecule tyrosine inhibitor anlotinib which developed in China has been approved as a third-line treatment for patients with small-cell lung cancer (SCLC). Our previous clinical study found that anlotinib combined with S-1 has better short-term ORR than the single-agent anlotinib of SCLC and other small-molecule vascular targeted drug therapies in the treatment of SCLC. However, the molecular mechanism of those effect remains unclear.
SCLC cell line H446 was treated with either anlotinib, 5-FU alone, or combination. The cellular effects including cell viability, cell apoptosis, cell cycle, cell migration, and invasion were explored to evaluate the cell proliferation level. Western blot was performed to determine the protein levels of the combined action of the two drugs. The xenograft mouse model was established by injection of H446 cells into mouse, and the animals were randomized and assigned for the drug treatments. Body weights and tumor sizes were recorded. WB was conducted using tumor tissues. All data were collected and statistically analyzed using -test to reveal the underlying molecular mechanism.
When anlotinib was combined with 5-FU, the IC50 value of cells was significantly reduced. And apoptosis, cell cycle arrest, and cell motility rates were stronger when anlotinib combined with 5-FU than in the anlotinib or 5-FU alone. In H446 cell-derived xenograft mouse model, tumor volumes were significantly decreased in Anlo/5-FU combination group than anlotinib or 5-FU alone group. Western blot showed the decreasing expression of p-Src/p-AKT in the Anlo/5-FU group.
Our data revealed that the treatment of combination of antitumor angiogenesis agent anlotinib with chemotherapy drug 5-FU may have synergistic cytotoxicity to SCLC in vitro and in vivo. This treatment modality reduced cell proliferation and migration via Src/AKT pathway. This new strategy may be a promising treatment for SCLC but needs to be confirmed in future clinical trials.
中国研发的小分子酪氨酸激酶抑制剂安罗替尼已被批准用于小细胞肺癌（SCLC）三线治疗。我们之前的临床研究发现，与安罗替尼单药或其他小分子血管靶向药物治疗 SCLC 相比，安罗替尼联合替吉奥（S-1）具有更好的短期 ORR。然而，这些疗效的分子机制尚不清楚。
采用安罗替尼、5-FU 单药或联合用药处理 SCLC 细胞系 H446，检测细胞活力、细胞凋亡、细胞周期、细胞迁移和侵袭等细胞增殖水平的变化，用 Western blot 检测两药联合作用的蛋白水平。将 H446 细胞注入小鼠建立异种移植瘤模型，随机分组给药，记录动物体重和肿瘤大小，用 Western blot 检测肿瘤组织蛋白水平。采用 t 检验对所有数据进行收集和统计学分析，以揭示潜在的分子机制。
当安罗替尼与 5-FU 联合使用时，细胞的 IC50 值明显降低，且与安罗替尼或 5-FU 单药相比，联合用药时细胞凋亡、细胞周期阻滞和细胞迁移率更强。在 H446 细胞来源的异种移植瘤小鼠模型中，与安罗替尼或 5-FU 单药组相比，安罗替尼联合 5-FU 组肿瘤体积明显减小。Western blot 显示，安罗替尼联合 5-FU 组的 p-Src/p-AKT 表达减少。
我们的数据表明，抗血管生成药物安罗替尼与化疗药物 5-FU 联合治疗可能具有协同的体外和体内 SCLC 细胞毒性作用。这种治疗方式通过 Src/AKT 通路减少细胞增殖和迁移。这种新的治疗策略可能是治疗 SCLC 的一种有前途的方法，但需要在未来的临床试验中得到证实。
