Kumari Rani, Rawat Kavita, Kumari Anupma, Shrivastava Anju
Department of Zoology, University of Delhi, Delhi, India.
Tumour Biol. 2017 Jun;39(6):1010428317705758. doi: 10.1177/1010428317705758.
For tumor to grow beyond 1-2 mm size, tumor recruits new blood vessels referred as angiogenesis; therefore, targeting angiogenesis can be a promising strategy to suppress cancer progression. In this study, in order to develop a good angiogenesis model, we investigated effect of Dalton's lymphoma on angiogenesis and further monitored the role of melatonin on regulation of angiogenesis. To evaluate angiogenesis, endothelial cells were isolated from main thoracic aorta and cultured in vitro in the presence or absence of Dalton's lymphoma supplemented with or without melatonin to monitor their role on its proliferation and migration, a hallmark of angiogenesis. Chick chorioallantoic membrane as well as mice mesentery which allows in vivo studies of tumor angiogenesis and testing of anti-angiogenic molecules was used to validate the in vitro analysis. To further extend our understanding about the regulation of the angiogenesis, we evaluated expression of tissue inhibitor of metalloproteinases 3, vascular endothelial growth factor, vascular endothelial growth factor receptor, and fibroblast growth factor in Dalton's lymphoma cells and mesentery by semiquantitative and quantitative reverse transcription polymerase chain reaction analysis. Dalton's lymphoma ascites induced significant increase in endothelial cell proliferation, migration, and sprouting of the tertiary branching in chorioallantoic membrane and mesentery of Dalton's lymphoma-bearing mice, whereas melatonin treatment led to their inhibition in a dose-dependent manner. Semiquantitative and quantitative reverse transcription polymerase chain reaction analysis of melatonin-treated Dalton's lymphoma cells and mesentery tissue clearly demonstrated restoration of angiogenesis-related genes tissue inhibitor of metalloproteinases 3 and reduction of vascular endothelial growth factor, vascular endothelial growth factor receptor, and fibroblast growth factor messenger RNA expression. Taken together, our results strongly demonstrate that Dalton's lymphoma provides pro-angiogenic environment leading to significant increase in angiogenesis, and further melatonin treatment reduced the Dalton's lymphoma ascites-induced angiogenesis implying that Dalton's lymphoma can serve as a very good model to study angiogenesis as well as for screening of drugs that can target angiogenesis.
为了使肿瘤生长超过1-2毫米大小，肿瘤会募集新的血管，即所谓的血管生成；因此，靶向血管生成可能是抑制癌症进展的一种有前景的策略。在本研究中，为了建立一个良好的血管生成模型，我们研究了道尔顿淋巴瘤对血管生成的影响，并进一步监测褪黑素在血管生成调节中的作用。为了评估血管生成，从胸主动脉分离内皮细胞，并在有或无道尔顿淋巴瘤且添加或不添加褪黑素的情况下进行体外培养，以监测其对内皮细胞增殖和迁移的作用，而增殖和迁移是血管生成的一个标志。鸡胚绒毛尿囊膜以及小鼠肠系膜可用于体内研究肿瘤血管生成和测试抗血管生成分子，以此来验证体外分析结果。为了进一步拓展我们对血管生成调节的理解，我们通过半定量和定量逆转录聚合酶链反应分析，评估了道尔顿淋巴瘤细胞和肠系膜中金属蛋白酶组织抑制剂3、血管内皮生长因子、血管内皮生长因子受体和成纤维细胞生长因子的表达。道尔顿淋巴瘤腹水可显著增加内皮细胞增殖、迁移以及荷道尔顿淋巴瘤小鼠的绒毛尿囊膜和肠系膜中三级分支的出芽，而褪黑素处理则以剂量依赖的方式导致这些现象受到抑制。对经褪黑素处理的道尔顿淋巴瘤细胞和肠系膜组织进行的半定量和定量逆转录聚合酶链反应分析清楚地表明，血管生成相关基因金属蛋白酶组织抑制剂3得以恢复，而血管内皮生长因子、血管内皮生长因子受体和成纤维细胞生长因子信使核糖核酸的表达则降低。综上所述，我们的结果有力地证明，道尔顿淋巴瘤提供了促血管生成环境，导致血管生成显著增加，而进一步的褪黑素处理减少了道尔顿淋巴瘤腹水诱导的血管生成，这意味着道尔顿淋巴瘤可作为研究血管生成以及筛选可靶向血管生成的药物的良好模型。
