ECV304-SKOV3和L_(929)-H_(22)细胞协同形成血管样结构的体外研究

In vitro study on synergism of ECV304-SKOV3 and L_(929)-H_(22) cells in tumor angiogenesis

目的:在建立肿瘤基质血管内皮细胞模型(ECV304-SKOV3)和成纤维细胞模型(L929-H22)的基础上,进一步探讨二者在肿瘤新生血管形成中的协同作用。方法:对ECV304-SKOV3和L929-H22细胞用RT-PCR测定其端粒酶活性,双室联合培养测定ECV304-SKOV3细胞游走与管腔形成能力、L929-H22细胞促进肿瘤细胞侵袭与内皮细胞管腔形成能力。结果:ECV304-SKOV3细胞和L929-H22细胞的端粒酶活性明显强于亲代细胞ECV304、L929,分别为0.778±0.011、0.875±0.026、0.692±0.014、0.684±0.012(P<0.001)。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),且管腔形成能力明显增强,管腔数多0.94倍(P<0.01),管腔长0.91倍(P<0.01)。L929-H22明显促进肿瘤细胞侵袭(P<0.001)与ECV304、ECV304-SKOV3细胞形成管型(P<0.05),尤其与ECV304-SKOV3联合培养时形成的管腔最多且最长(P<0.01;P<0.001)。结论:肿瘤基质血管内皮细胞存活能力、游走与管腔形成能力增强。肿瘤基质成纤维细胞明显促进肿瘤细胞侵袭与血管内皮细胞形成管型。二者在肿瘤新生血管形成中有协同作用。

Objective：To explore the synergism of vascular endothelial cells and fibroblasts derived from tumors in the tumor angiogenesis after the establishment of the cell models. Methods ： Reverse transcription polymerase chain reaction, two - well co culture system were used to detect the tolomerase activity, and their abilities of migration and angiogenesis of ECV304 - SKOV3 cells and the potential of L929 - H22 to promote angiogenesis and invasion of MDA MB 231 cells, respectively, which were established as the cell models for vascular endothelial cells and fibroblasts derived from tumors in our laboratory before. Results ： Tolomerase activity level of ECV304 SKOV3 cells was higher than that of ECV304 cells （0. 778 ± 0.011 vs 0. 692 ± 0. 014, P 〈 0. 001 ）, ,so were L929 - H22 cells and L929 cells （0. 875 ± 0. 026 vs 0. 684 ± 0. 012, P 〈 0.001 ）. The migratory capability of ECV304 SKOV3 cells was 3.10 times that of ECV304 cells（ P〈 0. 001 ）. ECV304 -SKOV3 cells formed tubular structures more and longer than ECV304 calls did when co-cultured with SKOV3 cells（ P 〈 0.01 ）. L929 822 and L929 cell lines enhanced the invasiveness of human mammary cancer MDA MB 231 cells through artificial basement membrane （Matrigel） 1.21 and 0.48 respectively（P〈 0. 001）. Fibroblasts L929-H22 stimulated endothelial cells to form more and longer tubes than L929 did（ P〈 0.05 ）, especially when L929 822 and ECV304 -SKOV3 cells were co - cultured. Conclusion ： The results suggest that vascular endothelial cells and fibroblasts derived from tumors survive longer with greater ability to migrate and form tubes, and promote tumor cells invasion and vascular endothelial cells to form tubes, The two kinds of cells have a synergic role in the tumor angiogenesis.