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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24h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12h和24h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[12h和24h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[12h和24h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。
The purpose of this study was to investigate the effect of tumor conditioned medium (TCM) on the proliferation, adhesion and migration of human umbilical vein endothelial cells (HUVECs). The MTT assay was used to determine the proliferation of endothelial cells after 24h treatment. The control group, TCM stock solution, TCM monodilution, TCM didilution, (N = 6). The fibronectin Fn-coated plates were used to detect the adhesion of endothelial cells to the basement membrane. The experiment was divided into control group and TCM untreated group (n = 6) The migration ability of cells was divided into control group and TCM untreated group (n = 6), and tested at 12h and 24h respectively. The results showed that different concentrations of TCM had different effects on the proliferation of endothelial cells. Compared with the control group (absorbance value of 0.58 ± 0.04), TCM stock solution had no significant effect on endothelial cell proliferation (absorbance value was 0.55 ± 0.01), TCM Both doubling and double dilutions significantly increased endothelial cell proliferation (absorbance values of 0.66 ± 0.03 (P <0.01) and 0.70 ± 0.02 (P <0.001), respectively]; TCM significantly decreased Compared with the control group [(14.05 ± 6.25) μm and (48.75 ± 16.37) μm] cells in the control group (12 h and 24 h, respectively), TCM significantly enhanced the migration ability [12 h and 24 h The migration distances were (68.25 ± 26.20) μm and (119.70 ± 34.90) μm, respectively, P <0.05]. The above results indicate that the inhibition of TCM on vascular endothelial cell adhesion and the enhancement of migration ability may be one of the important mechanisms of TCM in promoting tumor metastasis.