Study On The Effect And Regulation Of Monoacylglycerol Lipase On Non-small Cell Lung Cancer

Monoacylglycerol lipase(MGLL)is a serine hydrolytic enzyme that plays an important role in catalyzing the hydrolysis of Monoacylglycerol esters into glycerol and fatty acids,MGLL affects the malignant phenotypes of cancer cells,such as proliferation and invasion,by regulating the signaling pathway of lipid metabolism.In this paper,the role of MGLL in non-small-cell lung cancer(NSCLC)and its regulatory mechanism were investigated.1.Expression and Function of MGLL in NSCLC H460To explore the role of MGLL in lung cancer cells,firstly,the expression of MGLL in lung cancer cells A549 and H460 was detected by qRT-PCR and Western blot,results showed that the mRNA and protein expressions of MGLL in A549 and H460 were higher than those in normal lung epithelial cells HBE.When the expression of MGLL was silenced by si RNA,the ability of proliferation,migration,invasion and clone formation were significantly inhibited in NSCLC cell H460,the cells were arrested in the G0/G1 phase,the cells in the S and G2 phases were reduced,and the cell apoptosis was increased,with the expression of Bcl-2 protein were decreased after MGLL knockdown.In addition,the ability of tumorigenesis in vivo was inhibited after MGLL knockdown.Up-regulation of MGLL expression in H460 cells by transfected with the constructed MGLL-p IRES2-Zs Green1 vector could promote the ability of cell proliferation,migration,invasion and clone formation,but had no effect on cell cycle and apoptosis.These results suggest that MGLL act as oncogene in non-small cell lung cancer cell H460,and inhibition of MGLL may promote cell apoptosis by down-regulating Bcl-2.2.Verify the regulation of mi R-3200-5p and MGLLTo further determine the regulation of MGLL in lung cancer cells,by using bioinformatics analysis,miR-3200-5p was predicted to target 3’-UTR of MGLL mRNA.Results showed that miR-3200-5p was down-regulated in H460 and A549,the overexpression of mi R-3200-5p significantly inhibited the ability of proliferation,migration,invasion and plate clone formation,the cells were arrested in the G0/G1 phase,and cell apoptosis increased,while the overexpression of MGLL could attenuate the inhibitory effects caused by up-regulated of miR-3200-5p,the above results suggested that miR-3200-5p and MGLL might be negatively correlated.To clarify the potential regulation between miR-3200-5p and MGLL,dual-luciferase report assay vectors with MGLL WT 3’-UTR and MGLL Mut 3’-UTR were constructed,respectively.The transfection results showed that luciferase activity was inhibited with the wild-type reporter vector,and not affected in the mutant one,indicating that miR-3200-5p regulates gene expression by binding to MGLL 3’-UTR.In conclusion,MGLL is highly expressed in non-small-cell lung cancer cell,down-regulation of MGLL inhibited the ability of cell migration,invasion,cell clone formation and tumorigenesis in vivo.MGLL was the target gene of miR-3200-5p,overexpression of MGLL and miR-3200-5p could attenuate the inhibitory effect caused by up-regulated of miR-3200-5p on the malignant phenotype.Therefore,MGLL and miR-3200-5p can be considered as potential targets in the diagnosis and treatment of non-small-cell lung cancer.