Study On Mechanism Of LncRNA LINK-A Promoting Proliferation And Invasion Of Glioma Cells

Objective: To observe the expression and clinical significance of ln c RNA LINK-A in gliomas,and to explore the signaling mechanism of lncRNA LINK-A on the proliferation and invasion of glioma cells.Methods:1.A total of 88 cases of grade I-IV gliomas and adjacent tissues fr om neurosurgery of the Second Affiliated Hospital of Hebei Medical Un iversity from 2014 to 2016 were collected.RTFQ-PCR was used to det ect the expression of lncRNA LINK-A in glioma tissues and adjacent ti ssues.Immunohistochemical staining was used to analyze the expression of c-myc in glioma tissues and adjacent tissues.Pearson method was u sed to analyze the correlation between lncRNA LINK-A and c-myc in g lioma tissue.Log-rank test compared the 3-year overall survival of patie nts with low and high expression of lncRNA LINK-A and c-myc.2.Using glioma U251 cells as the research object,lncRNA LINKA over-expressing cell lines(LINK-A OV)and no-load cell lines((NC)were constructed by lentiviral transfection.Normal cells were also set as blank controls(Blank).f necessary,cells were pretreated with Rhosi n activity inhibitor Rhosin or Rock inhibitor GSK429286 A.MTT metho d was used to detect the cell viability,and crystal violet staining was u sed to investigate the cell proliferation ability.Invasion chamber experim ent was used to detect the cell invasion ability.Pull-Down test was use d to detect the Rho A activity,and western blot was used to detect the expression of Rho A,Rock1,c-myc,and Akt.Results:1.Clinical data show that the expression of lncRNA LINK-A is up-regulated in glioma tissues(P <0.01).Compared with grade ?+ ?glioma,lncRNA LINK-A was more highly expressed in grade ?+ ?g lioma(P <0.01).c-myc was weakly positive in adjacent tissues,positive in ?+ ?grade glioma tissues,and strongly positive in ?+ ?gra de glioma tissues.There was a strong positive correlation between lnc R NA LINK-A and c-myc in glioma tissues(r = 0.7545,P <0.01).Comp ared with patients with low expression,patients with high expression of lncRNA LINK-A or c-myc had lower overall survival rates at 3 years(P <0.01).;2.In vitro studies showed that overexpression of lncRNA LI NK-A significantly enhanced the proliferation and invasion ability of U251 cells(P <0.01).Overexpression of lncRNA LINK-A can enhance the activity of Rho A protein,and at the same time up-regulate the expressi on of Rock1,c-myc and Akt proteins(P <0.01).Pretreatment with Rho sin(2 ?M)or GSK429286A(0.5 ?M)can significantly reduce cell proli feration and invasion activity induced by overexpression of lncRNA LIN K-A(P <0.01).Rhosin pretreatment could reduce the increase of Rho A protein activity and up-regulation of Rock1,c-myc and Akt proteins in duced by lncRNA LINK-A overexpression(P <0.01).GSK429286 A pret reatment significantly inhibited the up-regulation of c-myc and Akt prote in expression induced by lncRNA LINK-A overexpression(P <0.01).Conclusions:1.The expression of lncRNA LINK-A and c-myc is increased in g lioma tissues,and it is related to tumor grade and poor prognosis.2.lncRNA LINK-A may up-regulate c-myc and Akt protein express ion through the Rho A / Rock1 signaling pathway,and promote the proli feration and invasion of glioma cells.