The Expression Of ZFAS In Colorectal Cancerand Itsrolein Tumormetastasis

Part?The expression and clinical significance of ZFAS1 in colorectal cancerAim:To measurethe expression of ZFAS1 in colorectal cancer(CRC)tissues and to analyze itsrelationship with clinicopathological parameters and the prognosis of CRC patients.Methods:Real time quantitative reverse transcription polymerase chain reaction(q RT-PCR) was usedtotestthe expression of ZFAS1 in 159 paired CRCtissues and adjacent noncancerous tissues(NCTs), and in 12 samples of hepatic metastases.Chi square testwas used to analyze the relationship between the ZFAS1 expression and clinicopathological parameters.Survival curve analyses were performedusingthe Kaplan-Meier method with the log-rank test applied for comparison. The Cox proportional hazards regression model was used to estimate the influence of ZFAS1 on the relapse and survialof CRC.Results:The expression of ZFAS1 in CRC tissues wasconspicuouslyhigher than that in their NCTs(P<0.0001), and 79 CRC tissuesshowed more than two-fold expression of ZFAS1 compared with corresponding NCTs.Moreover, the expression of ZFAS1 in hepatic metastases tissues was significantly higher than that observed in their primary sites(P<0.05). ZFAS1 expression in CRC tissues was positively correlated with lymphatic invasion(P=0.045) and TNM stage(P=0.003). Patients with increased ZFAS1 expression showed shorter relapse-free survival(P=0.012)and overall survival(P=0.003)of CRC patients. Cox multivariate analyses confirmed that ZFAS1 expression is an independent prognostic factor fortumor relapse(HR=2.132, 95% CI=1.015-4.546, P=0.048) and death(HR=1.878, 95% CI=1.006-3.529, P=0.049) of CRC.Conclusion: The expression of ZFAS1 is positively correlated with lymphatic invasion and TNM stage and appears to be a new prognostic factor for the relapse and death of CRC patients.Part? The role of ZFAS1 in the cellmigration and invasion of CRCAim:To studythe effectsof ZFAS1 in cell proliferation?migration and invasion of CRC cells.Methods:The expression of ZFAS1 m RNA in CRC cell lines was detected using q RT-PCR and two cell strains with higher ZFAS1 expressionwere chosen for the subsequent analyses. Si RNA technique was applied tosilence ZFAS1 expression;CCK-8 assay wasused to verify the influence of ZFAS1 on the CRC cell proliferation;Wound-healing assay and transwell assay were used to detect the effects of ZFAS1 on the migration and invasion ability of CRC cells. Molecular cloningtechniqueswereused to construct ZFAS1 sh RNA expression vector, and lentivirus production and transduction were used to generate sh ZFAS1 stablyexpressed CRC cell lines. Lung metastasis model in nude mouse was performed to detect the effects of ZFAS1 silencing on thein vivometastasis ability of CRC cells.Results:Silencing ZFAS1 expression has no obvious effect on the proliferation of CRC cells.Wound-healing assay and transwell assayshowed that silencing ZFAS1 expression inhibitedthe in vitro migration and invasion of CRC cells.Lung metastasis model in nude mouserevealedthat ZFAS1 knock downinhibitedthe in vivo metastasis of CRC.Conclusion:ZFAS1 plays important role in prompting migration, invasion and metastasis of CRC cells.Part? The primary mechanism of ZFAS1 in prompting CRC metastasisAim:To study the primary mechanism of ZFAS1 in CRC metastasis.Methods:QRT-PCR and Westernblot were used todetect the expression of EMT-related markers in ZFAS1-silenced CRC cells. Si RNA technique was applied tosilence ZEB1 expression, and to validate the relationship of ZEB1 with E-cad and VIM. Dual luciferase experiment was uesd to verify whether ZEB1 is the target genes of mi R-150-5p in CRC cells. Wound-healing assay and transwell assay were used to detect the effects of mi R-150-5p and ZEB1 on the migration and invasion ability of CRC cells.The relationship between ZFAS and mi R-150-5p was evaluated by luciferase experiment, wound-healing assay and transwell assays.Results:Epithelial cells marker CDH1(E-cad)was upregulated, whereas mesenchymal cells markers Vimentin(VIM) and ZEB1 were downregulated in ZFAS1-silenced CRC cells.ZEB1 could regulate the expression of E-cad and VIM. Mi R-150-5p inhibited CRC cell migration and invasion through targeting ZEB1. ZFAS1 blockedthe tumor suppressor function of mi R-150-5p by binding with it.Conclusion:ZFAS1 increased the expression of ZEB1, accelerated the procdess of EMT, and promoted the metastasis of tumor by inhibitingmi R-150-5p.