基于小鼠背脊皮翼视窗的肿瘤微环境活体可视化研究

In vivo visualization of tumor microenvironment based on mouse dorsal skin-fold window chamber

目的:构建小鼠背脊皮翼视窗(DSFC)模型,对肿瘤微环境进行持续动态监测,研究肿瘤增殖、扩散以及提供一种评估抗肿瘤药物的技术手段。方法:利用转染绿色荧光蛋白的小鼠乳腺癌4T1细胞构建小鼠DSFC肿瘤模型,对肿瘤微环境内肿瘤尺寸、血管形变、血流灌注量等参数进行实时动态连续监测,获得肿瘤微环境内的精确信息。同时评估纳米基因治疗药物PCL-PDEM-siRNA的治疗效果。结果:在裸鼠DSFC模型接种4T1乳腺癌细胞的成瘤率可达90%以上。肿瘤接种后2 d即可见接种区域毛细血管增多,4 d后可观察到边界清晰的肿瘤形态,第4~8天可见肿瘤区域血流灌注量显著提高。纳米药物注射4 d后肿瘤区域增长速度减缓,出现明显的肿瘤抑制效应,8 d后肿瘤内部新生血管减少,血流灌注量趋于稳定。结论:本研究为肿瘤微环境的精细化成像提供了新的策略,对肿瘤生物学的基础研究和抗肿瘤药物研发具有重要意义。

Objective To construct a mouse dorsal skin-fold window chamber(DSFC)model for monitoring the tumor microenvironment continuously,researching tumor proliferation and spread,and providing a technical method to evaluate the efficacy of anti-tumor drugs.Methods The mouse 4T1 breast cancer cells transfected with green fluorescent protein were used to construct a mouse DSFC tumor model for the real-time dynamic and continuous monitoring of tumor size,vascular deformation,blood perfusion and other parameters in the tumor microenvironment,thereby obtaining the precise tumor microenvironment information.Meanwhile,the proposed model was also used for evaluating the therapeutic effect of PCL-PDEM-siRNA in nanogene therapy.Results The tumorigenesis rate of 4T1 breast cancer cells inoculated in the DSFC model of nude mice was higher than 90%.Two days after tumor inoculation,the capillaries in the inoculated area were increased.On the 4th day after tumor inoculation,the tumor morphology with clear boundaries could be observed,and the blood perfusion volume at the tumor area was increased significantly on the 4th to 8th days after tumor inoculation.Four days after nanomedicine injection,the growth rate of the tumor area slowed down,and a significant tumor suppressor effect appeared;and 8 days after nanomedicine injection,the neovascularization inside the tumor was decreased,and the blood perfusion volume was beginning to stabilize.Conclusion The study provides a new strategy for the refined imaging of the tumor microenvironment,and is of great significance to the basic research of tumor biology and the development of anti-tumor drugs.