Seeliger Hendrik, Camaj Peter, Ischenko Ivan, Kleespies Axel, De Toni Enrico N, Thieme Susanne E, Blum Helmut, Assmann Gerald, Jauch Karl-Walter, Bruns Christiane J
Munich University Medical Center, Department of Surgery, Munich, Germany.
Mol Cancer Res. 2009 Feb;7(2):189-98. doi: 10.1158/1541-7786.MCR-08-0132. Epub 2009 Feb 10.
The progression of pancreatic cancer is dependent on local tumor growth, angiogenesis, and metastasis. EFEMP1, a recently discovered member of the fibulin family, was characterized with regard to these key elements of pancreatic cancer progression. Differential gene expression was assessed by mRNA microarray hybridization in FG human pancreatic adenocarcinoma cells and L3.6pl cells, a highly metastatic variant of FG. In vivo orthotopic tumor growth of EFEMP1-transfected FG cells was examined in nude mice. To assess the angiogenic properties of EFEMP1, vascular endothelial growth factor (VEGF) production of tumor cells, endothelial cell proliferation and migration, and tumor microvessel density were analyzed in response to EFEMP1. Further, tumor cell apoptosis, cell cycle progression, and resistance to cytotoxic agents were quantitated by propidium iodide staining and flow cytometry. In microarray hybridization, EFEMP1 was shown to be significantly up-regulated in L3.6pl cells compared with FG cells. Concordantly, EFEMP1 transfection of FG cells stimulated orthotopic and metastatic tumor growth in vivo. EFEMP1 expression resulted in a stimulation of VEGF production by tumor cells and an increased number of CD31-positive microvessels. Endothelial cell proliferation and migration were not altered by EFEMP1, indicating an indirect angiogenic effect. Further, EFEMP1 expression decreased apoptosis and promoted cell cycle progression in response to serum starvation or exposure to gemcitabine, 5-fluorouracil, and irinotecan. EFEMP1 has protumorigenic effects on pancreatic cancer in vivo and in vitro mediated by VEGF-driven angiogenesis and antiapoptotic mechanisms. Hence, EFEMP1 is a promising candidate for assessing prognosis and individualizing therapy in a clinical tumor setting.
胰腺癌的进展取决于局部肿瘤生长、血管生成和转移。EFEMP1是纤连蛋白家族最近发现的成员，已针对胰腺癌进展的这些关键要素进行了表征。通过mRNA微阵列杂交在FG人胰腺腺癌细胞和FG的高转移变体L3.6pl细胞中评估差异基因表达。在裸鼠中检查了EFEMP1转染的FG细胞的体内原位肿瘤生长。为了评估EFEMP1的血管生成特性，分析了肿瘤细胞的血管内皮生长因子（VEGF）产生、内皮细胞增殖和迁移以及肿瘤微血管密度对EFEMP1的反应。此外，通过碘化丙啶染色和流式细胞术对肿瘤细胞凋亡、细胞周期进程和对细胞毒性药物的抗性进行了定量。在微阵列杂交中，与FG细胞相比，EFEMP1在L3.6pl细胞中显著上调。与此一致，FG细胞的EFEMP1转染刺激了体内原位和转移性肿瘤的生长。EFEMP1表达导致肿瘤细胞VEGF产生增加和CD31阳性微血管数量增加。EFEMP1未改变内皮细胞的增殖和迁移，表明其具有间接血管生成作用。此外，EFEMP1表达降低了凋亡，并在血清饥饿或暴露于吉西他滨、5-氟尿嘧啶和伊立替康时促进了细胞周期进程。EFEMP1在体内和体外对胰腺癌具有促肿瘤作用，由VEGF驱动的血管生成和抗凋亡机制介导。因此，EFEMP1是临床肿瘤环境中评估预后和个体化治疗的有希望候选者。
