上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

存活素基因过表达对血管内皮细胞迁移和增殖能力的影响

陈江龙，张灏龙，赵渝，王学虎   

  1. 重庆医科大学 附属第一医院血管外科, 重庆 400016
  • 出版日期:2016-04-28 发布日期:2016-05-26
  • 通讯作者: 王学虎, 电子信箱: 184037696@qq.com。
  • 作者简介:陈江龙（1988—）， 男， 硕士生; 电子信箱: 843176155@qq.com。
  • 基金资助:

    国家自然科学基金青年科学基金项目（81200230）

Effects of survivin gene over-expression on migration and proliferation of vascular endothelial cells

CHEN Jiang-long, ZHANG Hao-long, ZHAO Yu, WANG Xue-hu   

  1. Department of Vascular Surgery, The First Affiliated Hospital of Chongqing Medical University， Chongqing 400016， China
  • Online:2016-04-28 Published:2016-05-26
  • Supported by:

    Young Scientists Project of National Natural Science Foundation， 81200230

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摘要：

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。

关键词: 存活素, 内皮细胞, 极光激酶B, 着丝粒内蛋白, 染色体过客蛋白, 有丝分裂

Abstract:

Objective To investigate changes in proliferation and migration in vascular endothelial cells (VECs) with over-expressed survivin (Svv) gene. Methods Rat aorta abdominalis segments were harvested and stable VECs were obtained by primary cell culture. Cell types were identified by observing cell shapes and using CD31 immunochemistry. VECs were assigned to three groups, i.e. blank control group (bc group), negative control group (nc group), and experiment group (svv group). The bc group used the serum free basic medium. The nc group and the svv group were transfected with empty virus vector and adenovirus carrying Svv gene, respectively. Changes of the Svv protein level in three groups were detected by Western blotting. The proliferation and migration of endothelial cells were measured by CCK8 assay and Transwell test, respectively. Aurora B, phosphorylated Aurora B, and Incenp levels were detected by Western blotting. Results The Svv protein level (P=0.000), proliferation, and migration of rat VECs were increased after being transfected with adenovirus carrying  Svv gene. Aurora B (P=0.000) and Incenp (P=0.000) levels and phosphorylation level of Aurora B (P=0.000) in the svv group were significantly increased as compared with the bc group and the nc group. Conclusion Over-expression of Svv gene can improve the proliferation and migration of rat VECs.

Key words: survivin, endothelial cells, Aurora B, Incenp, chromosomal passenge complex, mitosis