Baikun Li1,2, Ting Zhu1,3, Xiaohong Wu1,2, Shiyu Chen1,2, Chen Lu4, Jimin Zhu5,6, Qinglin Li7,8. 1. Key Laboratory of Xin'an Medicine, (Anhui University of Chinese Medicine), The Ministry of Education, Hefei, 230038, China. 2. School of Traditional Chinese Medicine, Anhui University of Chinese Medicine, Hefei, 230012, China. 3. School of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China. 4. School of Life Sciences, Anhui University of Chinese Medicine, Hefei, 230012, China. 5. Key Laboratory of Xin'an Medicine, (Anhui University of Chinese Medicine), The Ministry of Education, Hefei, 230038, China. zhjmcdc@126.com. 6. School of Life Sciences, Anhui University of Chinese Medicine, Hefei, 230012, China. zhjmcdc@126.com. 7. Key Laboratory of Xin'an Medicine, (Anhui University of Chinese Medicine), The Ministry of Education, Hefei, 230038, China. liqinglin@ahtcm.edu.cn. 8. School of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China. liqinglin@ahtcm.edu.cn.
Abstract
PURPOSE: We previously showed that the crosstalk of H1975 cells and platelets (PLTs) may promote tumor angiogenesis. This study aimed to determine whether other lung cell lines (LC) interacting with PLTs could affect tumor angiogenesis through in vivo and in vitro experiments. METHODS: Cell Counting Kit-8, EdU cell proliferation, wound healing, Transwell invasion, F-actin staining, tube formation, ELISA and western blot assays were performed to investigate the properties and the expression levels of vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2), p-VEGFR2, PI3K, p-PI3K, Akt, p-Akt and eNOS in supernatants or HUVECs. Then, using mouse models, immunohistochemistry was applied to detect the expression levels of CD31 and VEGF. RESULTS: Compared with single-cultured HUVECs (EC) or incubation with either LC supernatant (EC + LC) or activated PLT supernatant (EC + PLT), incubation with SN_LCP (supernatant derived from LC cocultured with PLT, named the EC + LC + PLT group) improved the viability, proliferation, migration, invasion, and tube formation activities of HUVECs and the expression of F-actin, VEGF, VEGFR2, p-VEGFR2, p-PI3K, p-Akt and eNOS in HUVECs. Higher expression levels of CD31 and VEGF were found in the LLC + PLT (mouse model inoculated with Lewis lung cancer (LLC) cells cocultured with PLTs) group than in the LLC (mouse model inoculated with LLC cells alone) group. However, the increased angiogenic properties of HUVECs were inhibited by apatinib, an inhibitor of VEGFR2. CONCLUSION: Lung carcinoma cells interacting with PLTs may play a key role in lung carcinoma angiogenesis through the VEGF/VEGFR2 signaling pathway.
PURPOSE: We previously showed that the crosstalk of H1975 cells and platelets (PLTs) may promote tumor angiogenesis. This study aimed to determine whether other lung cell lines (LC) interacting with PLTs could affect tumor angiogenesis through in vivo and in vitro experiments. METHODS: Cell Counting Kit-8, EdU cell proliferation, wound healing, Transwell invasion, F-actin staining, tube formation, ELISA and western blot assays were performed to investigate the properties and the expression levels of vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2), p-VEGFR2, PI3K, p-PI3K, Akt, p-Akt and eNOS in supernatants or HUVECs. Then, using mouse models, immunohistochemistry was applied to detect the expression levels of CD31 and VEGF. RESULTS: Compared with single-cultured HUVECs (EC) or incubation with either LC supernatant (EC + LC) or activated PLT supernatant (EC + PLT), incubation with SN_LCP (supernatant derived from LC cocultured with PLT, named the EC + LC + PLT group) improved the viability, proliferation, migration, invasion, and tube formation activities of HUVECs and the expression of F-actin, VEGF, VEGFR2, p-VEGFR2, p-PI3K, p-Akt and eNOS in HUVECs. Higher expression levels of CD31 and VEGF were found in the LLC + PLT (mouse model inoculated with Lewis lung cancer (LLC) cells cocultured with PLTs) group than in the LLC (mouse model inoculated with LLC cells alone) group. However, the increased angiogenic properties of HUVECs were inhibited by apatinib, an inhibitor of VEGFR2. CONCLUSION: Lung carcinoma cells interacting with PLTs may play a key role in lung carcinoma angiogenesis through the VEGF/VEGFR2 signaling pathway.
Authors: Majida Al-Abboodi; Ran An; Maximilian Weber; Rafael Schmid; Anne Klausing; Raymund E Horch; Anja M Boos; Annika Kengelbach-Weigand Journal: Oncol Rep Date: 2019-05-02 Impact factor: 3.906