肿瘤基质血管内皮细胞血管形成能力研究

Study on the angiogenic ability of tumor vascular endothelial cells

目的:探讨肿瘤基质血管内皮细胞的血管形成能力。方法:对卵巢癌细胞SKOV3培养上清液诱导人内皮细胞ECV304获得能稳定生长的ECV304-SKOV3细胞,用免疫细胞化学测定其血管形成因子的表达、PT-PCR测定其端粒酶活性、双室联合培养测定其游走能力与管腔形成能力,并分析其蛋白质合成能力、粘贴能力。并与亲代细胞ECV304比较。结果:ECV304-SKOV3细胞蛋白质合成能力、端粒酶活性强于亲代细胞ECV304(P<0.001),但粘贴能力减弱(P<0.001)。表达bcl-2、基质金属蛋白酶-9(matrixmetalloproteinase9,MMP-9)增强(P<0.05),而粘合素(tenascin,TN)减弱(P<0.05)。ECV304-SKOV3细胞较ECV304细胞的游走能力强2.10倍,差异非常显著(P<0.001),管腔形成能力也明显增强,平均每视野管腔数分别为(12.4±1.517)条和(6.4±0.894)条(P<0.01),管腔长分别为(262.17±49.52)μm和(137.34±18.69)μm(P<0.01)。结论:肿瘤基质血管内皮细胞存活能力增强,表达多种细胞因子与血管形成因子增强,相应地游走能力与管腔形成能力明显增强。

Objective:To explore the angiogenic ability of vascular endothelial cells in solid tumors.Methods:Immunocytochemistry,reverse transcription polymerase chain reaction (PT-PCR),and two-well co-culture system were used to detect the expressions of matrix metalloproteinase 9(MMP-9),transforming growth factor-β1(TGF-β1),tenascin(TN)and bcl-2,telomerase activity,and their ability of both migration and tube formation of ECV304-SKOV3 cells respectively,which ,as the cell model,had been established in our laboratory before.Then their abilities of protein synthesis and adhesion were analysed.Results:The ability to synthesize proteins and telomerase activity was enhanced(P<0.001).Adhesion potential of ECV304-SKOV3 cells decreased (P<0.001).The expressions of MMP-9 and bcl-2 in ECV304-SKOV3 cells increased (P<0.05),while TN decreased (P<0.05),as compared with that of ECV304 cells.The migratory capability of ECV304-SKOV3 cells was 2.10 times greater than that of ECV304 cells (P<0.001). ECV304-SKOV3 cells formed tubular structures more and longer than did ECB304 cells,when co-cultured with SKOV3 cells (P<0.01).Conclusion:The results suggest that ECV304-SKOV3 cells,compared with parental cells ECV304,survive longer,express and secrete various cytokines and angiogeneic factors stronger,and has greater ability to migrate and form vascular structure.