3 years ago

The regulation and mechanism of a dual PI3K/mTOR signaling inhibitor in hemangioma vascular endothelial cells in vitro

wangshu liu, Yang Yu, Juan Li, Hui Huang, Hao Liu, Meijia Yuan, Hongyang Tan, Nankai Wang, Miaojie Nie, Xuanshen Hu, Song Yu

Objective To observe the influence of the dual PI3K/mTOR inhibitor NVP-BEZ235 on proliferation and apoptosis of hemangioma cells in vitro and key molecules of the PI3K/Akt/mTOR signaling pathway.

Methods Hemangioma-derived endothelial cells (HeECs) were obtained by surgical resection and cultured after the explants with the trypsin-digestion method. Fourth generation cells were cultured with serum-free medium for 24 hours. Then, the intervention group cells were added to the culture medium with 0.50 μM or 1.00 μM NVP-BEZ235. Cell proliferation was detected with CCK-8 assays, apoptosis was detected by flow cytometry, and PI3K, Akt, mTOR, and p70s6k protein levels were detected by Western blots. Then, the relationship between the phenotype of hemangioma vascular endothelial cells and the four proteins was analyzed.

Results the 0.50 μM and 1.00 μM NVP-BEZ235 groups were significantly lower (0.88±0.03 and 0.59±0.05, respectively) than the control group (1.10±0.02) (P<0.01). The rate of G0/G1 phase cells in the 0.50 μM and 1.00 μM NVP-BEZ235 group were higher than the control group (P<0.01). The total rates of apoptotic cells in the 0.50 μM and 1.00 μM NVP-BEZ235 groups were higher than the control group (2.77±1.23)% (P<0.01). The PI3K pathway related protein levels in the NVP-BEZ235 group were lower than control group (P<0.01).

Conclusion The PI3K/Akt/mTOR signaling pathway participates in hemangioma development. NVP-BEZ235 affected hemangioma vascular endothelial cells in vitro by regulating the PI3K/Akt/mTOR signaling pathway in a dose-dependent manner.

Publisher URL: https://www.researchsquare.com/article/rs-17546/v1

DOI: 10.21203/rs.3.rs-17546/v1

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