Li Baikun, Dong Xingyu, Zhu Jimin, Zhu Ting, Tao Xiaoxiao, Peng Daiyin, Li Qinglin
School of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China.
School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China.
Oncol Lett. 2021 Sep;22(3):676. doi: 10.3892/ol.2021.12937. Epub 2021 Jul 21.
Activated platelets (PLTs) participate in the regulation of tumor angiogenesis, and tumors can activate PLTs. Whether co-culture of lung carcinoma with PLTs improves the function of human umbilical vein endothelial cells (HUVECs) requires further investigation. The present study aimed to investigate the impact of H1975 cell crosstalk with PLTs on the proliferation, migration and tube formation of HUVECs. Following generation of cell-derived supernatants and construction of the co-culture system, Cell Counting Kit-8, flow cytometry, transmission electron microscopy and a meter for epithelial measurement were performed to detect the proliferative ability of HUVECs. Furthermore, the wound healing and Transwell migration assays were performed to detect the migratory ability of HUVECs. A tube formation assay was performed to assess angiogenesis, ELISA was applied to detect the content of vascular endothelial growth factor (VEGF) and western blotting was carried out to measure the expression levels of VEGF receptor 2 (VEGFR2) in HUVECs. Compared with single-cultured HUVECs (control), co-culture with H1975 cells and PLTs (Exp_HP) improved cell proliferation, increased the proportion of cells in the S-phase, destroyed the cell ultrastructure and decreased transepithelial electrical resistance in HUVECs. In addition, a higher relative migration rate, greater number of migrated cells, stronger tube-forming ability and increased expression of VEGF and VEGFR2 were detected in the Exp_HP group compared with the control group. The properties of HUVECs in Exp_H (co-cultured with H1975 cells) were similar to those in Exp_HP, but significantly weaker. Taken together, the results of the present study suggest that tumor cells interacting with PLTs may play an important role in tumor angiogenesis by affecting or mediating changes in the properties of vascular endothelial cells (VECs).
活化血小板(PLTs)参与肿瘤血管生成的调节,且肿瘤可激活血小板。肺癌与血小板共培养是否能改善人脐静脉内皮细胞(HUVECs)的功能尚需进一步研究。本研究旨在探讨H1975细胞与血小板相互作用对HUVECs增殖、迁移和管腔形成的影响。在生成细胞来源的上清液并构建共培养体系后,采用细胞计数试剂盒-8、流式细胞术、透射电子显微镜和上皮测量仪检测HUVECs的增殖能力。此外,进行伤口愈合和Transwell迁移试验以检测HUVECs的迁移能力。进行管腔形成试验以评估血管生成,应用酶联免疫吸附测定(ELISA)检测血管内皮生长因子(VEGF)的含量,并进行蛋白质印迹法检测HUVECs中VEGF受体2(VEGFR2)的表达水平。与单培养的HUVECs(对照组)相比,与H1975细胞和血小板共培养(Exp_HP组)可改善细胞增殖,增加S期细胞比例,破坏细胞超微结构并降低HUVECs的跨上皮电阻。此外,与对照组相比,Exp_HP组检测到更高的相对迁移率、更多的迁移细胞、更强的管腔形成能力以及VEGF和VEGFR2表达增加。Exp_H组(与H1975细胞共培养)中HUVECs的特性与Exp_HP组相似,但明显较弱。综上所述,本研究结果表明,肿瘤细胞与血小板相互作用可能通过影响或介导血管内皮细胞(VECs)特性的变化在肿瘤血管生成中发挥重要作用。