Biological effects of neuritin on vascular endothelial cells of non-small cell lung cancer and its mechanism

ZHANG Jianqing; ZHANG Heng; WANG Zhen; ZHAO Hui

doi:10.3969/j.issn.1009-0460.2019.01.002

Abstract：Objective To investigate the biological effects and mechanism of neuritin abnormal expression on pulmonary vascular endothelial cells in non-small cell lung cancer (NSCLC). Methods 5-10 patients with NSCLC underwent surgery in the Xinjiang Uygur Autonomous Region People's Hospital from September to December 2017 were selected to extract primary human non-small cell lung cancer vascular endothelial cells (NSCLC-VECs), the primary cultured vascular endothelial cells of lung cancer patients with pathologically confirmed NSCLC were identified by immunohistochemistry with CD34 and Factor Ⅷ. The expression levels of neuritin in NSCLC-VECs and human pulmonary microvascular endothelial cells (HPMECs) were detected by quantitative PCR (QPCR) and Western-blotting. NSCLC-VECs and HPMEC cells were constructed to knock down and over express neuritin cells. MTT, cell scratch, Transwell chamber and flow cytometry were used to detect the effects of neuritin on cell proliferation and metastasis, cell cycle and apoptosis. The morphological changes of cells after transfection were observed by scanning electron microscope. Results The expression of neuritin in NSCLC-VECs cells was significantly higher than that in HPMEC cells (P ＜ 0. 01). After overexpression of neuritin, the Notch1 and VEGFR protein and mRNA expression in HPMEC and NSCLC-VECs cells increased significantly (P ＜ 0. 01). Cell proliferation, scratch healing and migration in vitro were significantly enhanced compared with the negative control group (P ＜ 0. 05). Cell surface pseudo foot was obvious, and the adhesion between cells increased. At the same time, overexpression of neuritin promoted cell cycle progression and inhibited cell apoptosis of HPMEC and NSCLC-VECs (P ＜ 0. 001). Conversely, after interfering with si-neuritin expression in HPMEC and NSCLC-VECs cells, the Notch1, VEGFR protein and mRNA expression was significantly inhibited (P ＜0. 01). Under electron microscopy, the cell surface was smooth, the cell proliferation activity, scratch healing ability and migration ability in vitro were significantly lower than those in the negative control group (P ＜ 0. 01). HPMEC and NSCLC-VECs cell cycle arrested in G0/G1 phase, and apoptosis rate increased significantly (P ＜ 0. 001). Conclusion Neuritin may be a potential biomarker of NSCLC, and it play a certain biological role in the genesis and development of NSCLC.