Research On The Mechanism Of LanCL2 Regulating Glioblastoma Cell Invasion

Objective: It was confirmed that Lan CL2 can regulate glioblastoma cells,explain the activation mechanism of Lan CL2 and identify its interacting proteins,providing a preclinical basis for the development of new glioblastoma treatment.Materials and methods:Using si RNA to interfere with the expression of Lan CL2 in glioblastoma cells,and construct a Lan CL2 knockout glioblastoma cell line using the CRISPR/Cas9 system,through transwell invasion assay,3D invasion assay,invasive pseudopodia formation assay,thorn Barbed end assay and laser scanning confocal technique were used to analyze the effect of targeting Lan CL2 on the formation and activity of invasive pseudopods.In addition,by constructing a LCL-overexpressing glioblastoma cell line,the transformation of Lan CL2 was also demonstrated by transwell invasion assay,3D invasion assay,invasive pseudopod formation assay,spur formation assay and laser scanning confocal technique,and reversed-phase phosphoric acid mass spectrometry.Regulatory effect on the invasiveness of glioblastoma cells.Results: The Lan CL2 gene has copy number variation and high amplification in 27% of glioblastoma tissues(154 of 580 are positive),and patients with glioblastoma with high expansion level of Lan CL2 are detected.Short overall survival;55% of glioblastoma samples(17 of 31 positive results),increased expression of Lan CL2 protein;of which,Lan CL2 protein was found in 8 relapsed glioblastoma samples All were significantly high expression and correlated with recurrence of glioblastoma(logistic regression analysis p < 0.05).In addition,Marques(Myristoylated alanine-rich C-kinase substrate),MARCKSL1(MARCKS-like protein 1),Cortactin,Stathmin 1,Myosin-9,Rasassociated and pleckstrin homology domains-containing protein 1(RAPH1)and CDC42 effector protein 4(Proteins such as CDC42EP4)are involved in cytoskeletal assembly and migration,whereas proteins and nuclear functions such as YAP1,STAT3,TAF12,NCOR2,NUP214,EIF4 B,and NIF1 B are involved in transcription factor activation.Conclusion: Lan CL2 gene copy number variation is associated with prognosis in patients with glioblastoma,and Lan CL2 protein expression is associated with glioblastoma recurrence and glioma grade.Lan CL2 is involved in the regulation of glioblastoma cell invasion.