Apolipoprotein M Up Regulates Matrix Metalloproteinase-10 Expression And Promotes Proliferation,Invasion And Migration Of A549 Cells

Objective:To investigate the effect of apolipoprotein M(apoM)on proliferation,invasion and migration of non-small cell lung cancer(NSCLC)A549 and association with Matrix Metalloproteinase-10(MMP-10).Explore the role and possible mechanisms of apoM in NSCLC.Methods:1.A549 cells were infected with vacant lentivirus and lentivirus carrying apoM sequence,and A549 cells were used as the research object to construct an in vitro model.The experimental groups were divided into two groups:(1)a control group infected with empty lentivirus;(2)an overexpression experimental group infected with apoM lentivirus.2.After successful infection,the mRNA and protein expression levels of apoM in the overexpressed group and the control group were detected by real-time fluorescent quantitative PCR(qRT-PCR)and Western Blot.3.After successful infection,the overexpression group and the control group were tested for cell proliferation multiplication by CCK-8 assay,cell migration ability by scratch test,and cell invasion ability by Transwell chamber invasion assay.4.The mRNA and protein expression levels of MMP-10 in the overexpression group and the control group were detected by qRT-PCR Western blotting technique.5.Statistical analysis of the data was performed using GraphPad 5.0 software,and the two sets of data were compared using an independent t test.p<0.05 was considered statistically significant.Results:1.Immunofluorescence photographs showed that the fluorescence intensity of the experimental group was significantly stronger than that of the control group after infection with apoM lentivirus and empty lentivirus.2.The results of qRT-PCR and Western blot showed that the mRNA level of apoM in the experimental group was significantly higher than that in the control group(P<0.01),and the protein level was also significantly increased(P<0.001).3.In the cell function experiment,the cell proliferation multiplication was detected by CCK-8 test.The results showed that the cell proliferation ratio of the experimental group was significantly higher than that of the control group(P<0.001).The cell migration efficiency was detected by the scratch test.The results showed that the experimental group was blank compared with the control group.The visual field was significantly reduced,and the closure rate was faster,that is,the mobility of the experimental group was significantly higher than that of the control group(P<0.001).The cell invasion rate was detected by Transwell chamber invasion assay.The results of crystal violet staining showed that the invasive rate of the experimental group was significantly higher than that of the control group(P<0.001).4.The results of qRT-PCR and Western blot showed that the mRNA level of MMP-10 in the experimental group was significantly higher than that in the control group(P<0.01),and the protein level was also significantly higher than that in the control group(P<0.05).Conclusion:1.After transfection of apoM lentivirus and empty lentivirus,the immunofluorescence intensity,apoM mRNA and protein levels of the overexpression group were higher than those of the control group,and the apoM overexpressing cell line was successfully constructed.2.Cell function experiments showed that overexpression of apoM promoted proliferation,migration and invasion of A549 cells.3.Overexpression of apoM can up-regulate the mRNA and protein expression levels of MMP-10.