【摘要】 目的 探讨血小板反应蛋白-1(thrombospondin-1,TSP-1)对体外肿瘤微环境下的淋巴管内皮细胞(lymphatic endothelial cells, LECs)增殖、迁移和侵袭能力的影响。方法 通过猪胸导管插管消化的方法，原代分离、培养LECs;采用VEGFR-3对LECs进行鉴定；采用CCK8检测TSP-1对乳腺癌细胞与LECs共培养模型下的LECs增殖活性；采用EdU实验检测TSP-1对乳腺癌细胞与LECs共培养模型下的LECs增殖水平；采用transwell小室检测TSP-1对乳腺癌细胞与LECs共培养模型下的LECs迁移和侵袭情况。结果 采用VEGFR-3对培养的LECs进行鉴定，为典型的LECs;采用CCK8对LECs增殖活力检测显示：当浓度为1.5ug/mL～2.5ug/mL时，TSP-1能明显抑制LECs的增殖活力(P<0.05);采用EdU对LECs增殖检测显示：当浓度为1.5ug/mL～2.5ug/mL时，TSP-1能明显抑制LECs的增殖水平(P<0.05);采用transwell小室检测LECs迁移和侵袭显示：当浓度为1.5ug/mL～2.5ug/mLl时，TSP-1能明显抑制LECs的迁移和侵袭(P<0.05)。结论 TSP-1对肿瘤微环境下的LECs增殖、迁移和侵袭能力有抑制作用，并且与药物剂量相关(低浓度TSP-1抑制作用不明显，高浓度TSP-1抑制作用明显)。更多还原

【Abstract】 Objective To investigate the effects of thrombospondin-1(TSP-1) on the proliferation, migration, and invasion ability of lymphatic endothelial cells(LECs) in the tumor microenvironment in vitro. Methods Digest and isolate endothelial cells from the thoracic ducts of pigs for primary culture; VEGFR-3 was used to idenifty LCEs; CCK8 assay was used to detect the proliferation activity of LECs by TSP-1 in the co-culture model; EdU assay was used for testing the cell proliferation level of LECs by TSP-1 in the co-culture model; Transwell chamber was used for testing the migration and invasion of LECs by TSP-1 in the co-culture model. Results The cultured LECs were identified using VEGFR-3 as typical LECs; The detection of proliferation activity of LECs using CCK8 showed that when the concentration was between 1.5 ug/ml and 2.5 ug/ml, TSP-1 could significantly inhibit the proliferation activity of LECs(P<0.05); The proliferation detection of LECs using EdU showed that when the concentration was between 1.5 ug/ml and 2.5 ug/ml, TSP-1 could significantly inhibit the proliferation level of LECs(P<0.05); Transwell chamber detection of LEC migration and invasion showed that TSP-1 could significantly inhibit the migration and invasion of LECs at concentrations ranging from 1.5 ug/ml to 2.5 ug/ml(P< 0.05). Conclusions TSP-1 has an inhibitory effect on the proliferation, migration, and invasion ability of LECs in the tumor microenvironment, and is related to drug dosage(low concentration TSP-1 has no significant inhibitory effect, high concentration TSP-1 has a significant inhibitory effect).更多还原