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Crosstalk between H1975 tumor cells and platelets to induce the proliferation,migration and tube formation of vascular endothelial cells
Authors:Baikun Li  Xingyu Dong  Jimin Zhu  Ting Zhu  Xiaoxiao Tao  Daiyin Peng  Qinglin Li
Affiliation:1.School of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China;2.School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China;3.School of Life Sciences, Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China;4.Key Laboratory of Xin''an Medicine (Anhui University of Chinese Medicine), The Ministry of Education, Hefei, Anhui 230038, P.R. China
Abstract: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).
Keywords:lung carcinoma   platelet   vascular endothelial cells   angiogenesis   co-culture
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