C19orf12 Promotes A549 Cell Metastasis And Regulates Its Metabolic Reprogramming

Objective:To investigate the effect of C19orf12 on tumor biological function and metabolic reprogramming of non-small cell lung cancer A549 cell line.Methods:In the first part,A549 cell line knockdown C19orf12 were constructed,and the knockdown efficiency of C19orf12 was verified by Western blot and real-time RT-qPCR at protein and mRNA levels,respectively.Transwell migration assay and wound healing assay and animal experiment to verify the effect of C19orf12 on migration function of A549 cells in vitro and in vivo.The proliferation and colony formation assay were used to detect the effect of C19orf12 on the proliferation of A549 cells in vitro.A549 scramble and shC19orf12 cells were injected into the mice to investigate the effect of C19orf12 on tumorigenesis in vivo.The effect of C19orf12 on apoptosis of A549 cells was detected by FITC Annxin V staining.In the second part,the biological function of C19orf12 in A549 cells was detected by RNA-seq assay.The mRNA expression of oxidative phosphorylation genes in A549 scramble,shC19orf12-1 and shC19orf12-2 cells was detected by RT-qPCR.In the third part,The extracellular acidification rate(ECAR)and oxygen consumption rate(OCR)of A549 scramble,shC19orf12-1 and shC19orf12-2 cells were measured by a seahorse cell energy metabolism meter.The effect of C19orf12 on the utilization nutrients of A549 cell was verified by measuring glucose,lactic acid and glutamine in the culture medium.Results:(1)The A549 cell line knocked down C19orf12 was successfully constructed.The number of migrated cells and scratch migration rate of A549 shC19orf12-1 and shC19orf12-2 cells were significantly lower than that of control A549 scramble cells.Compared with the A549 scramble cells,the number of lung metastases formed and the lung tissue weight of the knockdown group A549 shC19orf12-1 and shC19orf12-2 cells were significantly decreased.Compared with the A549 scramblecells,the knockdown group did not affect the proliferation,colony formation,volume and weight of subcutaneous tumor formation and apoptosis rate of A549 cell.(2)GSEA database analysis showed that the expression of the tricarboxylic acid cycle and oxidative phosphorylation gene was increased in the knockdown group A549 shC19orf12-1 compared with the control A549 scramble cells.RT-qPCR results showed that compared with the A549 scramble cells the mRNA expression of oxidative phosphorylation genes in the A549 shC19orf12-1 and shC19orf12-2 cells was significantly increased.(3)Compared with the control A549 scramble cells,the ECAR was reduced and the OCR was enhanced in A549 shC19orf12-1 and shC19orf12-2 cells.And glucose consumption,lactic acid production and glutamine consumption in A549shC19orf12-1 and shC19orf12-2 cells were significantly reduced.Conclusion:1.C19orf12 promotes the migration ability of A549 cells in vitro and in vivo.2.C19orf12 does not affect the proliferation and apoptosis of A549 cells.3.C19orf12 inhibited the oxidative phosphorylation of A549 cells and promoted its glycolysis function,which promoted the utilization of glucose and glutamine by A549 cells.4.C19orf12 may be a potential target for tumor metabolism therapy.