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肿瘤条件培养基对人脐静脉内皮细胞增殖、黏附、迁移能力的调节

张婷, 蒋春雷^*

第二军医大学海医系军事航海医学教研室，上海 200433

摘要

本文旨在研究肿瘤条件培养基(tumor conditioned medium, TCM)对人脐静脉内皮细胞(human umbilical vein endothelial cell, HUVEC)增殖、黏附和迁移能力的影响。采用MTT法测定TCM作用24 h后内皮细胞的增殖水平，实验设对照组、TCM原液(TCM stock solution)处理组、TCM一倍稀释液(TCM monodilution)处理组以及TCM二倍稀释液(TCM didilution)处理组(n = 6)；用纤连蛋白Fn包被培养板以检测内皮细胞与基底膜的黏附能力，实验分为对照组和TCM原液处理组(n = 6)；应用划痕法检测内皮细胞的的迁移能力，实验分为对照组和TCM原液处理组(n = 6)，并且取12 h和24 h两个时间点分别进行检测。结果显示，不同浓度的TCM对内皮细胞增殖能力的影响程度不同，相对于对照组(吸光度值为0.58 ± 0.04)，TCM原液对内皮细胞增殖无明显影响(吸光度值为0.55 ± 0.01)，TCM一倍稀释液和两倍稀释液均能显著促进内皮细胞的增殖[吸光度值分别为0.66 ± 0.03 (P < 0.01)和0.70 ± 0.02 (P < 0.001)]；与对照组相比，TCM可以显著降低内皮细胞与基底膜的黏附；与对照组[12 h和24 h细胞的迁移距离分别为(14.05 ± 6.25) µm和(48.75 ± 16.37) µm]相比较，TCM可显著增强其迁移能力[12 h和24 h的迁移距离分别为 (68.25 ± 26.20) µm和(119.70 ± 34.90) µm，均P < 0.05]。以上结果表明，TCM对血管内皮细胞黏附能力的抑制和迁移能力的增强可能是其促进肿瘤转移的重要机制之一。

关键词： 肿瘤条件培养基; 内皮细胞; 增殖; 黏附; 迁移

分类号：Q28

[Tumor conditioned medium regulates the proliferation, adhesion and migration of human umbilical vein endothelial cells.] [Article in Chinese]

ZHANG Ting, JIANG Chun-Lei^*

Department of Nautical Medicine, the Second Military Medical University, Shanghai 200433, China

Abstract

The aim of the present study was to investigate the effects of tumor conditioned medium (TCM) on the proliferation, adhesion and migration of human umbilical vein endothelial cell (HUVEC). Cells were divided into control group, TCM stock solution group, TCM monodilution group and TCM didilution group (n = 6), and MTT assay was used to determine endothelial cells proliferation level after being stimulated with TCM for 24 h. Coating culture dishes with Fn was used to detect the adhesion ability of endothelial cells to basement membrane in control group and TCM stock solution group (n = 6). Wound closure model was used to test the migration ability of endothelial cells in control group and TCM stock solution group at 12 h and 24 h (n = 6). The results showed that TCM at different concentrations influenced proliferation of endothelial cells in distinct degree. Compared to the control group (absorbance value: 0.58 ± 0.04), the stock solution had no effect on proliferation of endothelial cells (absorbance value: 0.55 ± 0.01). However, the TCM monodilution and the TCM didilution enhanced the proliferation of endothelial cells significantly (absorbance value: 0.66 ± 0.03, P < 0.01 and 0.70 ± 0.02, P < 0.001 separately). TCM downregulated the adhesion of endothelial cells to basement membrane. Moreover, compared to the migration distance of control group at 12 h and 24 h [(14.05 ± 6.25) µm and (48.75 ± 16.37) µm separately], TCM upregulated the basement membrane migration [(68.25 ± 26.20) µm and (119.70 ± 34.90) µm at 12 h and 24 h, both P < 0.05 separately]. These results suggest that TCM downregulates the adhesion ability of endothelial cells and upregulates their migration ability, and thus facilitates tumor metastasis.

Key words: tumor conditioned medium; endothelial cell; proliferation; adhesion; migration

收稿日期：2011-01-21　　录用日期：2011-03-24

通讯作者：蒋春雷　　E-mail: cljiang@vip.163.com

引用本文：

张婷, 蒋春雷. 肿瘤条件培养基对人脐静脉内皮细胞增殖、黏附、迁移能力的调节[J]. 生理学报 2011; 63 (3): 256-260.

ZHANG Ting, JIANG Chun-Lei. [Tumor conditioned medium regulates the proliferation, adhesion and migration of human umbilical vein endothelial cells.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (3): 256-260 (in Chinese with English abstract).