TW-37 对肺癌细胞和血管生成的影响及其机制研究

TW-37 inhibits angiogenesis and lung cancer cell growth by targeting ERK/VEGF

摘要:

目的 研究TW-37 对肺癌细胞生长和血管生成的影响及其机制。方法 使用不同浓度的TW- 37 处理人微血管内皮细胞（HMEC-1）。采用qRT-PCR 和蛋白免疫印迹法检测细胞内血管内皮生长因子 （VEGF） 及其转录因子的转录和表达，通过血管生成与鸡胚绒毛尿囊膜（CAM） 试验检测HMEC-1 细胞的 血管生成能力，通过MTT 与流式细胞术分析细胞增殖、细胞周期和细胞凋亡情况。建立异种移植模型，检 测TW-37 对肿瘤进展的影响。处死小鼠并收集肿瘤，采用免疫组织化学法分析VEGF 和血管密度标志蛋白 的表达。结果 TW-37 可以抑制CAM 上生成新的血管，并且呈剂量依赖性。TW-37 可抑制HMEC-1 细胞 的迁移和侵袭（P <0.05），并诱导细胞凋亡（P <0.05）。TW-37 可抑制VEGF、ERK 及HIF-1α的表达（P < 0.05）。体内实验表明，TW-37 可以抑制肿瘤生长和血管生成。结论 TW-37 通过ERK/VEGF 抑制血管生 成和肺癌的体内外进展。

Abstract:

Objective To study the effect and mechanism of TW-37 on lung cancer cell growth and angiogenesis. Methods After treatment of human microvascular endothelial cells (HMEC-1) with different concentrations of TW-37, the transcription and expression levels of vascular endothelial growth factor (VEGF) and its transcription factors in the cells were detected by polymerase chain reaction (PCR) and immunoblotting. The chick embryo chorioallantoic membrane (CAM) assay was performed to detect the angiogenic ability of HMEC-1 cells. The cell proliferation, cell cycle and apoptosis were analyzed by MTT assay and flow cytometry. The xenograft model was established and the effect of TW-37 on tumor progression was detected. Then the mice were sacrificed and tumor tissues were collected, where the expression of VEGF and marker proteins of vascular density was analyzed by immunohistochemistry. Results TW-37 significantly inhibited the formation of new blood vessels on CAM in a dose-dependent manner (P < 0.05). TW-37 also suppressed the migration and invasion of HMEC-1 cells (P < 0.05), and induced apoptosis through cell cycle arrest (P < 0.05). TW-37 could inhibit the expression of VEGF, extracellular signal-regulated kinase (ERK) and hypoxia inducible factor-1α (HIF-1α) (P < 0.05). Besides, in vivo experiments showed that TW-37 could inhibit tumor growth and angiogenesis. Conclusions TW-37 inhibits angiogenesis and lung cancer progression in vivo and in vitro through ERK/VEGF.