目的：观察干扰乳腺癌MDA-MB-231细胞中期因子( midkine, MK)表达对内皮细胞的增殖、迁移、脉管形成能力的变化,分析MK在肿瘤血管形成中的作用。方法采用shRNA法降低人乳腺癌MDA-MB-231细胞MK的表达,通过RT-PCR及Western blot法检测MK干扰效果,实验分为MK干扰组、空载体组及对照组；采用制备肿瘤条件培养基模拟肿瘤微环境培养HUVECs细胞,通过CCK-8法检测各组内皮细胞增殖能力、Transwell小室检测内皮细胞迁移能力、Matrigel检测内皮细胞脉管形成能力。结果与对照组及空载体组相比,MK干扰组细胞的增殖、迁移及脉管形成能力均明显降低(P<0．05)。结论干扰乳腺癌MDA-MB-231细胞MK的表达可抑制内皮细胞增殖、迁移、脉管形成能力,提示MK可能在肿瘤血管形成中起重要作用。%Purpose To observe the effect of MK on proliferation, migration and angiogenesis of human umbilical vein endothelial HU-VECs cells and to explore the role of MK in tumor angiogenesis. Methods siRNA targeting MK was used to downregulate MK expres-sion in human breast cancer cell line MDA-MB-231. Then, the experiment was divided into three groups: the untreated group, the empty vector transfection group and MK gene interference group. CCK-8 assay was used to detect the endothelial cells proliferation, tr-answell method was for detection of endothelial cell migration numbers, and matrigel in vitro small tube formation assay was used to sur-vey the state of tube formation. Expression of MK in siRNA transfection was identified by RT-PCR and Western blot. Results The MK gene interference group showed lower cell proliferation activity, less number of migration of cells and tube formation than the other two groups (P<0. 05). Conclusion Low-expressed MK in human breast cancer cell line MDA-MB-231 can inhibit proliferation, mi-gration and angiogenesis of endothelial cells, which shows that MK may play an important role in tumor angiogenesis.
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