The Role And Mechanism Of LncRNA ZEB1-AS1 In Gastric Cancer

PURPOSESZinc finger E-box binding homeobox 1 antisense 1(ZEB1-AS1)is a long non-coding RNA,which has found to upregulated in various kinds of cancer.However,the role of the lncRNA ZEB1-AS1 in gastric cancer is not thoroughly understood.This research was conducted to demonstrate the association between ZEB1-AS1 expression levels and clinical outcome or prognosis of cancer patients and investigate its role in the biological behavior of gastric cancer cells.METHODS1.Five databases were retrieved by using keywords and included eligible researches according to inclusion and exclusion criteria.After data extraction and study quality assessment,Meta-analysis,statistical analysis and online database analysis were conducted to demonstrate the relationship between ZEB1-AS1 expression and clinicopathological factors cancer patients.2.ZEB1-AS1 expression in gastric cancer tissues and the relationship between survival rate and the expression of ZEB1-AS1 was investigated through public cancer dataset.The expression of ZEB1-AS1 in human gastric cancer cell lines were also examine by qRT-PCR.3.ZEB1-AS1 specific siRNA were transfected into MGC-803 cells and the expression of ZEB1-AS1 was enhanced in AGS cell line by transfection with GV144-ZEB1-AS1.4.Cell Counting Kit-8(CCK-8)assay and colony formation assay were performed to observe the proliferation of gastric cancer cells after transfection of GV144-ZEB1-AS1 and si-ZEB1-AS1.The transwell migration assay was used in MGC-803 and AGS cells after transfection.5.Western blot was performed to examine the expression change of proliferation,metastasis and EMT indicators in MGC-803 and AGS cells after transfection.6.Western blotting for nuclear/cytoplasmic protein and cellular immunofluorescence to investigate the possible potential involvement of signaling pathways in ZEB1-AS1-mediated EMT and cell migration.RESULTS1.18 studies were included in this study by retrieving 5 databases.Overexpression ofZEB1-AS1 is significantly correlated with poor OS(HR=2.24,95% CI = 1.86-2.86,P<0.0001).ZEB1-AS1 expression levels were also associated with clinicopathological parameters including lymph node metastasis(Yes vs.No;OR=4.80,95%CI=3.11-7.43,P<0.00001),histologic differentiation(Moderate + poor vs.Well;OR=2.72,95% CI: 1.69-4.37,P<0.0001),tumor metastasis and invasion(Yes vs.No;OR=2.52,95%CI=1.21-5.68,P=0.03)and TNM stage(III+IV vs.I+II;OR=3.04,95% CI: 1.67-5.52,P=0.0003).2.ZEB1-AS1 expression is increased in gastric cancer tissues and associated with poor survival.It was highest in MGC-803 cell line and lowest in AGS cell line,which showed the significant difference compared with GES-1.3.Aberrant expression of ZEB1-AS1 could regulate proliferation and migration in gastric cancer cells through modulating the production of MMP2,MMP9 and PCNA.4.Increased ZEB1-AS1 expression promoted cell EMT process by up-regulation of ZEB1,Vimentin and Snail,whereas down-regulation of E-cadherin.5.ZEB1-AS1 could regulate Smad2/3-?-catenin-NF-?B signaling and promote the translocation of ?-catenin,p-Smad2/3 and p65 from the cytolplasm to nucleus CONCLUSIONThe results indicated potential value of ZEB1-AS1 as a biomarker for predicting a poor prognosis in patients with cancer.ZEB1-AS1 may also play an interaction role in Smad2/3-?-catenin-NF-?B signaling pathway to promote proliferation,metastasis and EMT in gastric cancer.