RESUMO
BACKGROUND: ADAMTS8 expression has been identified to be low in many cancers including lung cancer. However, the specific functions and regulatory system of ADAMTS8 remain to be unveiled. PURPOSE: To study the potential modulatory mechanism of ADAMTS8 in lung cancer in cell and xenograft mice models. METHODS: Differential expression of ADAMTS8 in lung cancer was analyzed on online tools. So was the overall survival curve in association with ADAMTS8/VEGFA expression in lung cancer patients. RT-qPCR was applied to validate the ADAMTS8 expression in lung cancer cell lines H460 and A549, with the normal lung epithelial cell Beas-2b as a control. Thereafter, overexpressed and knockdown plasmids were constructed for transfection. Colony and flow cytometry methods were used for cell proliferation and apoptosis. RT-qPCR and Western blot methods validated the changes in VEGFA after ADAMTS8 regulation in cells. Tube formation and Transwell methods were applied to observe the changes in tube formation and migration in HUVECs induced by tumor conditioned medium (TCM). Stable-transfected cells were injected subcutaneously into nude mice. H&E and Immunohistochemistry were applied to analyze the pathological differences and protein changes of ADAMTS8, VEGFA and CD31. RESULTS: High ADAMTS8 was correlated with high overall survival rate in lung cancer patients. ADAMTS8 was also abnormally downregulated in NSCLC cells. Upregulation of ADAMTS8 suppressed cell proliferation and enhanced apoptosis while downregulation of ADAMTS8 promoted cell proliferation and decreased apoptosis. VEGFA was negatively correlated with ADAMTS8 in lung cancer tissues. Upregulation of ADAMTS8 inhibited VEGFA in mRNA and protein levels. Further, knockdown of ADAMTS8 induced tube formation and migration of HUVECs and upregulation of ADAMTS8 inhibited this. In addition, upregulation of ADAMTS8 in nude mice inhibited tumor growth and also suppressed VEGFA and CD31 in tumors. CONCLUSION: ADAMTS8 inhibited lung cancer progression through suppressing VEGFA in lung cancer.
Assuntos
Proteínas ADAMTS/genética , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/patologia , Fator A de Crescimento do Endotélio Vascular/genética , Proteínas ADAMTS/metabolismo , Animais , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Células Endoteliais da Veia Umbilical Humana , Humanos , Neoplasias Pulmonares/genética , Camundongos Endogâmicos BALB C , Neovascularização Patológica/genética , Taxa de Sobrevida , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Carbon Fiber Reinforced Polymer (CFRP) can be used to reinforce steel structures depending on its high strength and lightweight resistance. To analyze and evaluate the load-carrying capacity of CFRP-reinforced steel structures. This study uses the Finite Element Analysis (FEA) and the experimental tests combined to investigate the influence that the reinforcement patterns and the relevant parameters have on the load-carrying capacity. We made specimens with different reinforcement patterns. Take the steel beam specimen with full reinforcement as an example. Compared with the load-carrying capacity of the steel beam reinforced by two-layer CFRP cloth, that respectively increases by 5.16% and 11.1% when the number of the CFRP cloth increases to four and six, respectively. Based on a specimen set consisting of CFRP-reinforced steel structures under different reinforcement patterns, the random forest algorithm is used to develop an evaluation model for the load carrying. The performance test results show that the MAE (Mean Absolute Error) of the evaluation model can reach 0.12 and the RMSE (Root Mean Square Error) is 0.25, presenting a good prediction accuracy, which lays a solid foundation for the research on the CFRP-based reinforcement technology and process.