Lysyl oxidase inhibition via βaminopropionitrile attenuates hypoxia induced migration and invasion of non-small cell lung cancer cells

Wang Kai; Wang Congjian; Ju Xuetao; Jiao Zhenhua; Yu Jun

doi:10.3969/j.issn.1671-6450.2024.10.002

Abstract：Objective To investigate the effect and mechanism of aminopropionitrile(βAPN)on hypoxia induced metastasis and invasion of non-small cell lung cancer(NSCLC)by inhibiting lysyl oxidase(LOX).Methods Experiments on the expression and catalytic activity of LOX in NSCLC cells under hypoxia state:A549 and SPCA1 cell lines were divided in-to normal oxygen group,hypoxia 12h group and hypoxia 24h group.Experiments on changes of NSCLC cell migration and in-vasion ability and epithelial mesenchymal transition(EMT)related molecular expression induced by βAPN mediated LOX in-activation:A549 and SPCA1 cell lines were divided into normal oxygen group,hypoxia group,hypoxia+βAPN group.LOX activity of the two cells was detected by fluorescence assay.The expression of hypoxia inducible factor-1 α(HIF-1α),LOX,epithelial cadherin(E-cadherin),neural cadherin(N-cadherin),matrix metallopeptidase 2(MMP-2)and matrix metallopeptidase 9(MMP-9)genes were detected by reverse transcription and quantitative real time PCR(qRT PCR).The protein expression levels of E cadherin,N cadherin,MMP-2 and MMP-9 were detected by Western blot.Transwell assay was used to detect the migration and invasion ability of NSCLC cells.Results Compared with normorxia group,the expression levels of HIF-1αmRNA and LOX mRNA in A549 and SPCA1 cells in hypoxia 12h and hypoxia 24h group were significantly up regulated(P＜0.01),HIF 1α protein and LOX protein were significantly up-regulated(P＜0.01),and LOX enzyme activity of were sig-nificantly increased(P＜0.01).Compared with normorxia group,the migration and invasion ability of A549 and SPCA1 cell lines in hypoxia group were enhanced,while all significantly decreased in hypoxia+βAPN group(P＜0.01).Compared with normorxia group,the expression level of E cadherin mRNA in hypoxia group was decreased,and the expression levels of N cadherin mRNA,MMP-2 mRNA and MMP-9 mRNA were increased,while the expression level of E cadherin mRNA in-creased,and the expression levels of N cadherin mRNA,MMP-2 mRNA and MMP-9 mRNA decreased in hypoxia+βAPN group(P＜0.01).Compared with normorxia group,the expression of E cadherin protein decreased and the expression of N cadherin,MMP2 and MMP9 protein increased in the hypoxia group,while the expression levels of N cadherin,MMP-2 and MMP-9 protein decreased,E cadherin protein was increased in hypoxia+βAPN group(P＜0.01).Conclusion βAPN can attenuate hypoxic induced migration and invasion of NSCLC by inhibiting LOX enzyme.