重组人血管内皮抑制素对肺微血管内皮细胞增殖及其周期分布的影响

目的：探讨重组人血管内皮抑制素（恩度）对肺微血管内皮细胞增殖的影响及机制。方法：以人非小细胞肺癌细胞系Ａ５４９和人肺微血管内皮细胞系ＨＰＭＥＣ建立非接触式共培养血管模型，并在此模型上应用ＭＴＴ方法研究肿瘤血管内皮细胞的增殖，以流式细胞仪检测血管内皮细胞的周期分布，以ＲＴ-ＰＣＲ和Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ检测血管内皮细胞的ｃｙｃｌｉｎＤ１转录水平和蛋白表达水平的变化。结果：恩度能明显抑制共培养模型下血管内皮细胞的增殖，并呈剂量依赖性；肿瘤内皮细胞Ｇ０／Ｇ１细胞数增多，Ｓ期细胞显著减少；转录和蛋白检测水平ｃｙｃｌｉｎＤ１呈剂量依赖性下调。结论：恩度能抑制肿瘤血管内皮细胞增殖，且这一作用与抑制ｃｙｃｌｉｎＤ１表达，使肿瘤血管内皮细胞滞留于Ｇ０／Ｇ１期有关。

Effect of endostar on proliferation and cell cycle of lung cancer microvasculature endothelial cell HPMEC

Objective： To study effect of recombinant human endostatin （endostar） on proliferation of lung cancer microvascular endothelial cells and its mechanism. Methods ： Non-contact co-culture system containing human pulmonary microvascular endothelial cells （HPMECs） and human non-small cell lung cancer cells A549 was established. On this model, the proliferation of HPMECs was determined by the method of MTF. Flow cytometry was used to analyze the change of cell cycle. Western blotting was preformed to determine cyclin D1. Results：The proliferation of HPMECs was inhibited by endostar in a dose dependent mode. Flow cytometry resuits showed that the percentage of G0/G1 phase cells was increased and that of S phase cells was significantly decreased by endostar compared with those of Untreated group. Endostar also down-regulated cyclin D1 expression. Conclusion：It was suggested that endostar can inhibit tumor microvasculature endothelial cells proliferation. The action was associated with arrest of G0/G1 and down-regulation of cyclin D1.