论文部分内容阅读
背景与目的:血管新生与肿瘤生长密切相关,烟曲霉素(Fumagillin)可特异性抑制血管内皮细胞的增殖,但其对结直肠癌的治疗作用尚不十分清楚。本研究旨在探讨Fumagillin对结直肠癌细胞系生长的抑制作用及其作用机制。方法:将WiDr或HT-29细胞以5×105/L的量分别接种于20只SCID小鼠背部皮下,接种4周后每2日腹腔注射Fumagillin(0.1mg/kg)或Cyclo(1mg/kg)(Arg-Gly-Asp-D-Phe-Val)持续4周,然后处死动物并测量原发瘤质量及原发瘤内微血管密度。在体外培养脐带静脉内皮细胞(HUVECs),向培养基内加入Fumagillin(0.01mg/kg)或Cyclo(0.1mg/kg),观察其对HUVECs增殖及微管形成的抑制作用。以基因芯片技术筛选Fumagillin处理后HUVECs的基因表达变化,并以定量PCR和免疫印迹技术对基因芯片的结果做进一步验证。结果:Fumagillin处理组小鼠的结直肠癌原发瘤质量及肿瘤内CD105阳性的微血管数量显著小于对照组小鼠(P<0.05),体外实验显示,Fumagillin处理后HUVECs的增殖和微管形成受到明显抑制,基因芯片检测显示Fumagillin处理后HUVECs有71个基因表达上调和143个基因表达下调,表达变化的基因多与细胞黏附、移动、增殖和基因转录有关。定量PCR和免疫印迹技术发现Fumagillin抑制HUVECs表达cyclinE2,白细胞活化黏附因子(activated leukocyte cell adhesion molecule,ALCAM)和细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)。结论:Fumagillin通过抑制新生血管的形成来抑制结直肠癌的生长。
BACKGROUND & OBJECTIVE: Angiogenesis is closely related to tumor growth. Fumagillin can specifically inhibit the proliferation of vascular endothelial cells. However, the therapeutic effect of Fumagillin on colorectal cancer is not yet clear. This study aimed to investigate the inhibitory effect of Fumagillin on the growth of colorectal cancer cell lines and its mechanism. METHODS: WiDr or HT-29 cells were inoculated subcutaneously in the back of 20 SCID mice at a dose of 5 × 10 5 / L. Fumagillin (0.1 mg / kg) or Cyclo (1 mg / kg) ) (Arg-Gly-Asp-D-Phe-Val) for 4 weeks, then the animals were sacrificed and the primary tumor mass and the primary intracranial microvessel density were measured. Umbilical vein endothelial cells (HUVECs) were cultured in vitro, and Fumagillin (0.01 mg / kg) or Cyclo (0.1 mg / kg) was added into the culture medium to observe the inhibitory effect on HUVECs proliferation and microtubule formation. The gene expression changes of HUVECs after Fumagillin treatment were screened by gene chip technology. The results of gene chip were further verified by quantitative PCR and Western blotting. Results: The tumor mass of primary colorectal cancer and the number of CD105 positive microvessels in Fumagillin-treated mice were significantly smaller than those in control mice (P <0.05). In vitro experiments showed that the proliferation and microtubule formation of HUVECs treated with Fumagillin were affected The results of gene chip showed that 71 genes were up-regulated and 143 genes were down-regulated in HUVECs after Fumagillin treatment. The genes that were changed were mostly associated with cell adhesion, migration, proliferation and gene transcription. Quantitative PCR and Western blotting showed that Fumagillin could inhibit the expression of cyclinE2, ALCAM and ICAM-1 in HUVECs. Conclusion: Fumagillin inhibits the growth of colorectal cancer by inhibiting the formation of new blood vessels.