The Expression Of AQP1 In Gliomas And The Effect On Its Biological Behavior

Objective The expression of AQP1 in human glioma tissues and its relationship with clinical pathological parameters and the prognosis were explored.Then AQP1 was successfully transfected into LN229 cells with lentivirus vector and the biological effects on tumor cells as well as underlying mechanisms were analyzed,which represent an active target site for therapeutic purpose.Methods 1.186 paraffin-embedded clinical glioma samples were collected from the patients of Tianjin Medical University Cancer Institute and Hospital,and 135 cases of patients with complete data were followed up.2.Immunohistochemistry was performed to detect the expression of AQP1 and β-catenin,and their correlation with pathological parameters,prognosis and the interrelationship were analyzed.3.Western blotting was conducted to test AQP1 expression in normal brain tissue,benign tumor and glioma tissues.4.We constructed recombinant lentivirus vector with m GFP label.PCDH-CMV-MCS-EF1-Puro-m GFP was used as a control group.AQP1 were spliced into the linear PCDH-CMV-MCS-EF1-Puro-m GFP vector,being as the experiment group.5.After packing the lentivirus of the recombinant plasmids by using HEK 293 T cells,LN229 cells were infected with them.Stable cells were screened by puromycin and named as AQP1/LN229,vector/LN229.6.Western blotting was applied to detect the expression of each protein and stable cell clones were screened.7.MTT assay was performed to detect the state of cell proliferation which was persistently cultured for 7 d.8.Brd U assay was performed to detect the proliferation ability of glioma cell.9.Soft agar assay was performed to detect the colony formation ability of glioma cell which was persistently cultured for 2 weeks.10.Transwell invasion assay was performed to detect the invasiveness ability in vitro in LN229 cells transfected with lentiviral.11.Western blotting assay was performed to detect the expression of β-catenin in AQP1/LN229 cells and control cells.Results 1.The expression of AQP1 was increased with the pathological grade(P<0.05),but with no relationship with age,gender,tumor size,peritumor edema,tumor location and pathological type(P>0.05).2.Western blotting showed that the expression of AQP1 was higher in high grade gliomas.3.Kaplan-Meier showed that AQP1 expression was negative correlated with overall survival and progression-free survival.4.Univariate survival analysis showed that AQP1,β-catenin,histological grade,tumor size and age were correlated with overall and progression-free survival.5.In multivariate survival analysis,AQP1 and histological grade were significant independent prognostic factors for overall survival and progression-free survival.6.AQP1 expression coincided with β-catenin expression in glioma samples.Spearman analysis showed a strong correlation between AQP1 and β-catenin expression.7.AQP1 expression in AQP1/LN229 cells were significantly higher than the control cells,so as to prove that the expression of AQP1 was stably overexpressed.8.In MTT assay,the proliferation capacity of AQP1/LN229 cells was obviously enhanced than the vector/LN229 cells(P<0.05).9.Brd U labeling assay showed that AQP1 group proliferated rapider than vector group(P<0.05).10.Soft agar assay showed that the colony formation ability of AQP1 overexpression group was obviously enhanced compared with the vector group(P<0.05).11.Transwell assay showed that after AQP1 lentiviral infection,the number of LN229 cell-penetrating significantly enhanced,higher than the control group.12.β-catenin expression was up-regulated in AQP1/LN229 cells(P<0.05).Conclusion 1.AQP1 expression was increased in glioma tissue and positively correlated with the pathologic stage and poor prognosis.2.AQP1 expression was positively associated with β-catenin in glioma tissue.3.AQP1 and histological grade were significant independent prognostic factors for overall survival and progression-free survival.4.When AQP1 in glioma cells was over-expressed,enhanced growth,motility and invasiveness were found.5.When AQP1 in LN229 was over-expressed,β-catenin expresssion were enhanced.