人类CRIPTO-1在肿瘤血管生成中的作用。

Role of human cripto-1 in tumor angiogenesis.

作者信息

Bianco Caterina, Strizzi Luigi, Ebert Andreas, Chang Cindy, Rehman Aasia, Normanno Nicola, Guedez Liliana, Salloum Rita, Ginsburg Erika, Sun Youping, Khan Nadia, Hirota Morihisa, Wallace-Jones Brenda, Wechselberger Christian, Vonderhaar Barbara K, Tosato Giovanna, Stetler-Stevenson William G, Sanicola Michele, Salomon David S

机构信息

Tumor Growth Factor Section, Mammary Biology and Tumorigenesis Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.

出版信息

J Natl Cancer Inst. 2005 Jan 19;97(2):132-41. doi: 10.1093/jnci/dji011.

DOI:10.1093/jnci/dji011

PMID:15657343

Abstract

BACKGROUND

Human cripto-1 (CR-1) promotes cell transformation and increases migration and invasion of various mouse and human epithelial cell lines. We investigated whether CR-1 also stimulates angiogenesis.

METHODS

We used human umbilical vein endothelial cells (HUVECs) to measure in vitro migration with fibronectin-coated Boyden chambers, invasion with Matrigel-coated Boyden chambers, proliferation with a tetrazolium salt, and differentiation with an in vitro Matrigel assay. We investigated new blood vessel formation in vivo by use of Matrigel-filled silicone cylinders implanted under the skin of nude mice and by use of a breast cancer xenograft model with CR-1-transfected or control Neo-transfected MCF-7 human breast cancer cells. We also used a blocking anti-CR-1 monoclonal antibody to investigate the role of CR-1 in angiogenesis in vivo and in vitro. All statistical tests were two-sided.

RESULTS

CR-1 stimulated HUVEC proliferation, migration, and invasion and induced HUVEC differentiation into vascular-like structures on Matrigel. In vivo, recombinant CR-1 protein induced microvessel formation in Matrigel-filled silicone cylinders, and microvessel formation was statistically significantly inhibited with a blocking anti-CR-1 monoclonal antibody (CR-1 and antibody = 127% of microvessel formation compared with that in untreated control cylinders and CR-1 alone = 259%; difference = 132%, 95% confidence interval [CI] = 123% to 140%; P<.001). Tumors formed by CR-1-transfected MCF-7 cells in the cleared mammary fat pad of nude mice had higher microvessel density than tumors formed by control Neo-transfected MCF-7 cells (CR-1-transfected cells = 4.66 vessels per field and Neo-transfected cells = 2.33 vessels per field; difference = 2.33 vessels per field, 95% CI = 1.2 to 2.8; P = .004).

CONCLUSION

CR-1 appears to have an important role in the multistep process of angiogenesis.

摘要

背景

人CRIPTO-1（CR-1）可促进细胞转化，并增加多种小鼠和人类上皮细胞系的迁移和侵袭能力。我们研究了CR-1是否也能刺激血管生成。

方法

我们使用人脐静脉内皮细胞（HUVECs），通过纤连蛋白包被的博伊登小室检测体外迁移能力，通过基质胶包被的博伊登小室检测侵袭能力，通过四唑盐检测增殖能力，通过体外基质胶试验检测分化能力。我们通过将填充有基质胶的硅胶柱植入裸鼠皮下以及使用转染了CR-1或对照Neo的MCF-7人乳腺癌细胞的乳腺癌异种移植模型来研究体内新血管形成情况。我们还使用一种阻断性抗CR-1单克隆抗体来研究CR-1在体内和体外血管生成中的作用。所有统计检验均为双侧检验。

结果

CR-1刺激HUVEC增殖、迁移和侵袭，并诱导HUVEC在基质胶上分化为血管样结构。在体内，重组CR-1蛋白可诱导填充有基质胶的硅胶柱内形成微血管，并且一种阻断性抗CR-1单克隆抗体可使微血管形成受到统计学显著抑制（CR-1与抗体组的微血管形成是未处理对照柱的127%，单独使用CR-1组为259%；差异为132%，95%置信区间[CI]=123%至140%；P<0.001）。在裸鼠清除乳腺脂肪垫中，转染CR-1的MCF-7细胞形成的肿瘤比对照Neo转染的MCF-7细胞形成的肿瘤具有更高的微血管密度（转染CR-1的细胞=每视野4.66条血管，转染Neo的细胞=每视野2.33条血管；差异=每视野2.33条血管，95%CI=1.2至2.8；P=0.004）。