Role And Mechanism Of GA6 In Glucose Metabolism In Cancer

Objective:Malignant tumors have become a major threat to health of human today.new problems have emerged With the emergence of new treatments.The mechanism of tumorigenesis,the problems of drug resistance and the development of new anticancer drugs should be solved urgently.Hypoxia inducible factor 1(HIF1)has powerful function in its active subunit HIF1?.Previously,we found that HIF1? interacted with GA6 by IR spectrum.As a newly discovered protein,GA6 has not been reported has interaction with HIF1 a.This study includes the interaction between GA6 and HIF1?.The effects of cell and tissue levels in vitro on the biological function and glycolysis key enzymes of GA6 and the relationship with drug resistance will provide a new direction for the adjuvant therapy of malignant tumors.Methods:In order to explore the relationship between GA6 and HIF1,we constructed the expression vector expressing GA6 and HIF1 a,and then transfection 293T cells to immunoprecipitation to determine the interaction,and purified the GST fusion protein expressed by GA6 mutants and performed GST pulldown experiment with HIF1?.The relationship of HIF1? to GA6 was determined.Then in cancer cell line,Western blot was used to sturdy the relationship between GA6 and HIF1? and the key enzymes of glycolysis key at the protein level,and RT-PCR was used to detect the relationship between HIF1? and the key enzymes of oxidative phosphorylation at the transcriptional level,and the experiment was repeated in vivo in mice.Finally,the relationship between GA6 and drug resistance was explored through the cultivation of resistant pemetrexed A549 strain.Results:The interaction between GA6 and HIF1 a was confirmed by immunoprecipitation,and it was further found that HIF1? binds to its 542-731 segment.By knockdown and overexpression,it was found that GA6 could promote the expression of HIF1 a at the protein and transcription levels.The biological function of GA6 was confirmed in knockout mice to promote the growth and regulation of glycolysis of tumor cells.The interaction with RAS showed us the possibility of drug resistance.Conclusion:In our experiment,we found a new tumor cell glucose metabolism regulator,GA6.GA6 has the function of regulating key enzymes of glycolysis from transcription,translation,and biological activities,etc.,because it has the ability to regulate HIF1 aand tumor glycolysis may be regulated by this way.The correlation between GA6 and drug resistance was demonstrated by the enhanced combination with k-ras G12D.