Study Of The Mechanism Of HIF-1?- Induced NDUFA4L2 Protein In The Hypoxia Microenvironment Of Lung Cancer

Objective: To explore the mechanism of NDUFA4L2 protein pathway induced by HIF1? in the development of non-small cell lung cancer(NSCLC)Methods:(1)The expression of mitochondrial NDUFA4L2 protein in non-small cell lung adenocarcinoma tissues and adjacent tissues was detected by real-time quantitative PCR(qPCR),Western blotting and Immunohistochemical staining.(2)Non-small cell lung cancer A549 and H1299 cell lines were cultured under normal and hypoxic conditions.HIF1? and NDFUA4 L 2 expression levels were detected by Western blotting.Moreover,the cells were treated with HIF1? inhibitor 2-ME and DMOG at different times(0h?24h?48h?72h)and different concentrations(0,2,4,8,16)?M.(3)Experiment of cell clone formation was used to detect cell proliferation,scratch test was used to detect cell migration,and flow cytometry was used to detect cell apoptosis.(4)Non-small cell lung cancer A549 and H1299 cells were cultured under normal and hypoxic conditions.NDUFA4L2 was silenced to observe the changes of HIF1? expression,phenotype and production of ROS of NSCLC cells.Results:(1)NDUFA4L2 protein was highly expressed in non-small cell lung cancer.(2)NDUFA4L2 protein was also overexpressed in NSCLC cell lines cultured under hypoxia.(3)The co-localization of HIF-1a and NDUFA4L2 protein was in mitochondria(4)The proliferation,cloning,migration and invasion of NSCLC cell lines were inhibited by 2-ME and promoted by DMOG.(5)Silencing NDUFA4L2 protein did not affect the expression of HIF-1?.(6)Through mitochondrial NDUFA4L2,HIF-1a regulated the phenotype of NSCLC cells under hypoxia.(7)Silencing NDUFA4L2 can promote ROS production in A549 cells cultured under hypoxia,rather than the cells cultured in normal oxygen.(8)Silencing NDUFA4L2 can promote apoptosis of NSCLC cell lines and inhibit EMT production.Conclusions: The anti-cancer effect of HIF-? can directly target the mitochondrial NDUFA4L2 protein.Therefore,it can hopefully become a new anti-cancer target.