Sola Francesco, Gualandris Anna, Belleri Mirella, Giuliani Roberta, Coltrini Daniela, Bastaki Maria, Tosatti Maria Pia Molinari, Bonardi Fabrizio, Vecchi Annunciata, Fioretti Francesca, Ciomei Marina, Grandi Maria, Mantovani Alberto, Presta Marco
Pharmacia-Upjohn, Nerviano, 20014 Milan, Italy.
Angiogenesis. 1997;1(1):102-116. doi: 10.1023/A:1018309200629.
Basic fibroblast growth factor (FGF-2) is expressed in vascular endothelium during tumor neovascularization and angioproliferative diseases, including vascular tumors and Kaposi's sarcoma (KS). We have investigated the in vivo biological consequences of endothelial cell activation by endogenous FGF-2 in a mouse aortic endothelial cell line transfected with a retroviral expression vector harboring a human FGF-2 cDNA and the neomycin resistance gene. FGF-2 transfectants, named pZipbFGF2-MAE cells, caused the rapid growth of highly vascularized, non-infiltrating tumors when injected in nude mice. In contrast, lesions grew poorly when cells were injected in immunocompetent syngeneic animals. Histologically, the tumors had the appearance of hemangioendothelioma with spindled areas resembling KS and with numerous CD31+ blood vessels and lacunae. Southern blot analysis of tumor DNA, as well as disaggregation of the lesion followed by in vitro cell culture, revealed that less than 10% of the cells in the tumor mass retain FGF-2 overexpression and neomycin resistance at 6-8 weeks post-injection. Nevertheless, in vitro G418 selection allowed the isolation from the tumor of a FGF-2-overexpressing cell population showing biochemical and biological characteristics similar to those of pZipbFGF2-MAE cells, including the capacity to originate vascular lesions when re-injected in nude mice. To evaluate the effect of angiostatic compounds on the growth and vascularization of pZipbFGF2-MAE cell-induced lesions, nude mice were treated weekly (100mg/kg, i.p.) with the angiostatic sulfonated distamycin A derivative 2,2'-(carbonyl-bis-[imino-N-methyl-4,2-pyrrole carbonyl-imino-{N-methyl-4,2-pyrrole}carbonylimino])-bis-(1,5-naphthalene) disulfonic acid (PNU 153429). The results demonstrate that PNU 153429 inhibits the growth of the lesions and causes a approximately 50% decrease in CD31+ microvessel density. In conclusion, the data indicate that FGF-2-overexpressing endothelial cells cause vascular lesions in immunodeficient mice which may represent a novel model for opportunistic vascular tumors suitable for the evaluation of angiostatic compounds.
碱性成纤维细胞生长因子（FGF-2）在肿瘤新生血管形成以及包括血管肿瘤和卡波西肉瘤（KS）在内的血管增殖性疾病的血管内皮中表达。我们研究了用携带人FGF-2 cDNA和新霉素抗性基因的逆转录病毒表达载体转染的小鼠主动脉内皮细胞系中内源性FGF-2激活内皮细胞的体内生物学后果。FGF-2转染细胞，命名为pZipbFGF2-MAE细胞，注射到裸鼠体内时会导致高度血管化、非浸润性肿瘤的快速生长。相反，当将细胞注射到具有免疫活性的同基因动物体内时，病变生长不良。组织学上，肿瘤具有血管内皮瘤的外观，有类似KS的梭形区域，有大量CD31+血管和腔隙。对肿瘤DNA进行Southern印迹分析，以及对病变进行解离后进行体外细胞培养，结果显示在注射后6-8周，肿瘤块中不到10%的细胞保留FGF-2过表达和新霉素抗性。然而，体外G418选择允许从肿瘤中分离出一个FGF-2过表达细胞群体，其显示出与pZipbFGF2-MAE细胞相似的生化和生物学特性，包括再次注射到裸鼠体内时引发血管病变的能力。为了评估血管生成抑制化合物对pZipbFGF2-MAE细胞诱导病变的生长和血管化的影响，每周（100mg/kg，腹腔注射）用血管生成抑制性磺化双咪唑霉素A衍生物2,2'-（羰基-双-[亚氨基-N-甲基-4,2-吡咯羰基-亚氨基-{N-甲基-4,2-吡咯}羰基亚氨基]）-双-（1,5-萘）二磺酸（PNU 153429）处理裸鼠。结果表明，PNU 153429抑制病变的生长，并使CD31+微血管密度降低约50%。总之，数据表明FGF-过表达的内皮细胞在免疫缺陷小鼠中导致血管病变，这可能代表了一种适合评估血管生成抑制化合物的机会性血管肿瘤的新模型。
