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目的研究过表达存活素(Svv)基因的血管内皮细胞增殖和迁移能力的变化。方法切取若干大鼠腹主动脉段,行原代细胞培养得到稳定生长的大鼠动脉内皮细胞株,通过观察细胞形态及CD31细胞免疫荧光检测明确细胞类型。将细胞分成3组,分别为空白对照组(bc)、阴性对照组(nc)及实验组(svv)。bc组用无血清基础培养基,nc组采用空病毒载体转染,svv组采用携带Svv基因的腺病毒转染。Western blotting法检测3组细胞Svv蛋白水平的变化,CCK8法检测内皮细胞增殖能力,Transwell实验检测细胞迁移能力。Western blotting法检测极光激酶B(Aurora B)、磷酸化Aurora B和内着丝粒蛋白(Incenp)的含量。结果携带Svv基因的腺病毒转染成功后大鼠动脉内皮细胞Svv蛋白表达量增高(P=0.000),迁移、增殖能力均提高。svv组细胞的Aurora B(P=0.000)、Incenp(P=0.000)含量和Aurora B的磷酸化水平(P=0.000)均较bc组和nc组显著提高。结论 Svv基因过表达可提高大鼠动脉内皮细胞的迁移和增殖能力。
Objective To study the changes of proliferation and migration of vascular endothelial cells over-expressed survivin (Svv) gene. Methods The abdominal aorta of some rats was excised and primary cultured cells were stably cultured in order to observe the cell morphology and CD31 immunofluorescence. The cells were divided into 3 groups: blank control group (bc), negative control group (nc) and experimental group (svv). Serum-free basal medium was used in group bc, nc group was transfected with empty vector, and svv group was transfected with adenovirus carrying Svv gene. Western blotting was used to detect the changes of Svv protein levels in three groups. CCK8 was used to detect the proliferation of endothelial cells. Transwell assay was used to detect the migration of cells. The contents of Aurora B, phosphorylated Aurora B and Incenp were detected by Western blotting. Results The expression of Svv protein was increased (P = 0.000) and the ability of migration and proliferation was increased after transfection of adenovirus carrying Svv gene. Aurora B (P = 0.000), Incenp (P = 0.000) and Aurora B phosphorylation (P = 0.000) in svv group were significantly higher than those in bc group and nc group. Conclusion Svv gene overexpression can improve the migration and proliferation of rat endothelial cells.