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Aim:To explore the mechanism of differentiated embryo-chondrocyte expressedgene 1 (DEC 1) in the lung adenocarcinoma A549 cell line and its relationship withhypoxia-inducible factor 1α(HIF-1α).Methods:DEC 1 and HIF-1α protein levelswere detected in lung adenocarcinoma tissues by immunohistochemistry.A549cells were cultured at hypoxia.MTT assay was used to determine the effect of cellviability.The apoptotic analysis was determined by flow cytometry.RT-PCR andimmunocytochemistry were carried out to observe the DEC 1 and HIF-1α mRNAand protein expression.HIF-1α mRNA and protein levels were detected in thestably transfected pcDNA-DEC 1+/-A549 cells by RT-PCR and Western blotting.DEC1 mRNA and protein levels were detected in the stably transfected pcDNA-DEC 1+/-A549 cells with or without H1F-1α antisense oligonucleotide treatmentby RT-PCR and Western blotting.Results:DEC1 was specifically located in 40(48.8%) lung adenocarcinoma tissues.The results of the MTT test showed thegrowth of the stably transfected pcDNA-DEC+ A549 cells was higher than thoseof A549 cells and the stably transfected pcDNA-DEC-A549 cells.Hypoxia alsoinduced the apoptosis of A549 cells and the stably transfected pcDNA-DEC+/-A549 cells.Hypoxia increased the mRNA and protein levels of DEC1 and HIF-1α.Both HIF-1α mRNA and protein levels decreased in the stably transfected pcDNA-DEC+cells,but HIF-1α antisense oligonucleotide had no effect on DEC1 in thestably transfected pcDNA-DEC+/-A549 cells.Conclusion:DEC 1 is a marker oftumor hypoxia.DEC1 downregulates the HIF-1α at both mRNA and protein levelsat hypoxia in lung adenocarcinoma A549 cells.
Aim: To explore the mechanism of differentiated embryo-chondrocyte expressed gene 1 (DEC 1) in the lung adenocarcinoma A549 cell line and its relationship with hypoxia-inducible factor 1α (HIF-1α). Methods: DEC 1 and HIF- lung adenocarcinoma tissues by immunohistochemistry. A549 cells were cultured at hypoxia. MTT assay was used to determine the effect of cellviability. apoptotic analysis was determined by flow cytometry. RT-PCR and immunocytochemistry were carried out to observe the DEC 1 and HIF-1α mRNA and protein expression.HIF-1α mRNA and protein levels were detected in the stably transfected pcDNA-DEC 1 +/- A549 cells by RT-PCR and Western blotting. DEC1 mRNA and protein levels were detected in the stably transfected pcDNA-DEC 1 +/- A549 cells with or without H1F-1α antisense oligonucleotide treatment by RT-PCR and Western blotting. Results: DEC1 was specifically located in 40 (48.8%) lung adenocarcinoma tissues. The results of the MTT test showed the growth of the stably transfected pcDNA-DEC + A549 cells were higher than those of A549 cells and the stably transfected pcDNA-DEC-A549 cells. Hypoxia also induced the apoptosis of A549 cells and the stably transfected pcDNA-DEC +/- A549 cells. Hypoxia increased the mRNA and protein levels of DEC1 and HIF-1α.Both HIF-1α mRNA and protein levels decreased in the stably transfected pcDNA-DEC + cells, but HIF-1α antisense oligonucleotide had no effect on DEC1 in the stably transfected pcDNA-DEC +/- A549 cells. Conlusion : DEC 1 is a marker of tumor hypoxia. DEC1 downregulates the HIF-1α at both mRNA and protein levelsat hypoxia in lung adenocarcinoma A549 cells.