LUC7L3 Affects The Proliferation And Invasion Of Malignant Meningioma Cells Through YAP1 Signaling

Objective:To investigate whether LUC7L3 affects the proliferation and invasion of malignant meningioma,and to explore whether LUC7L3 plays a role through the YAP1 signaling pathway.Methods:1.Malignant meningioma IOMM-Lee cell line was infected with lentivirus,and the expression level of LUC7L3 was down-regulated and up-regulated,respectively.The experiment was divided into five groups: blank group(blank),silent negative control group(NC-sh),silent LUC7L3 group(LUC7L3-sh),overexpression negative control group(NC-ox),overexpression LUC7L3 group(LUC7L3-ox).The expression levels of LUC7L3 mRNA and protein in each group of cells were detected by real-time fluorescent quantitative PCR and Western blot.2.Cell proliferation was detected by CCK8,plate cloning experiment and EdU proliferation experiment,and cell cycle changes were detected by flow cytometry.3.Cell migration and invasion were observed by cell scratch test and Transwell cell invasion test.4.Western blot was used to detect LUC7L3,YAP1,ERK1/2,p-ERK1/2,NF-kB,p-NF-kB,Vimentin,Elk1,p-Elk1,Caspase-3,p-S6,mTOR,p-mTOR,HER-2,PARP,PI3 K,Merlin protein expression levels.5.After using p-ERK1/2 specific inhibitor U0126,the expression levels of LUC7L3,YAP1,ERK1/2,p-ERK1/2,NF-kB,p-NF-kB protein in each group of cells were detected.Results:1.The expression level of LUC7L3 mRNA in malignant meningioma cells was detected by qPCR.LUC7L3 mRNA was expressed in malignant meningioma cells.The expression level of LUC7L3 mRNA was decreased in the silencing group,but increased in the overexpression group.Western blot method was used to detect the expression level of LUC7L3 protein in each group: the expression level of LUC7L3 protein in the silencing group decreased,while the expression level of LUC7L3 protein in the overexpression group increased.2.By observing the cell morphology of each group,the cell morphology of the silent group was significantly changed,while that of the overexpressed group was not.CCK8 cell proliferation assay showed that the proliferation capacity of the silent group decreased,while the overexpression group increased.The panel cloning experiment showed that the cloning rate decreased in the silence group,but increased in the overexpression group.The EdU proliferation assay showed that the proliferation capacity of the silent group decreased,while the overexpression group increased.The difference was statistically significant(P<0.05).Flow cytometry: LUC7L3 works by affecting G0/ g1-s in the cell cycle.3.The cell migration and invasion ability of the silent group decreased;while the cell migration and invasion ability of the overexpression group increased.4.Silent and overexpressed LUC7L3,the expression level of YAP1 protein also changed accordingly,the difference was statistically significant(P <0.05).In addition,the expression of p-ERK1 / 2 and p-NF-κB in the silent group increased unexpectedly,the difference was statistically significant(P <0.05),while there was no significant change in the overexpression group,the difference was not statistically significant(P> 0.05).p-ERK1/2,p-NF-kB,Vimentin,Elk1,p-Elk1,Caspase-3,p-S6,mTOR,p-mTOR,HER-2,PARP,PI3 K,Merlin protein expression did not change significantly No statistical significance(P> 0.05).5.After using U0126 to suppress the expression of p-ERK1/2 in IOMM-Lee cells,the expression level of p-ERK1/2 decreased significantly,while the expression of LUC7L3,YAP1,ERK1/2,NF-kB,p-NF-kB did not change significantly The difference was not statistically significant(P <0.05).Conclusion:1.LUC7L3 promotes the proliferation and invasion of malignant meningioma cells.2.LUC7L3 promoted the proliferation and invasion of meningioma cells through YAP1 signal.3.Silencing LUC7L3 may increase the expression of p-ERK1/2 through an alternative pathway.