ZNF382 Regulates Epithelial-mesenchymal Transition And Functions As A Tumor Suppressor Silenced By CPG Methylation In Gastric Cancer

Objective: Several studies recently reported that ZNF382(KRAB zinc finger protein 382)was downregulated in multiple carcinomas,due to its promoter methylation.Nonetheless,the exact role of ZNF382 in gastric carcinogenesis is still elusive.Thus,we studied the alterations and functions of ZNF382 in gastric carcinoma(GC)pathogenesis.Methods: Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR),quantitative real-time PCR(qPCR),and immunohistochemistry were carried out to detect its expression pattern in GC cell lines and gastric tissue samples.Furthermore,its methylation status in GC cell lines and tumor tissues was detected by methylation-specific PCR(MSP).Colony formation,CCK-8,flow cytometric analysis,wound healing and trans-well assay were performed in ZNF382-transfected and vector-transfected GC cell lines to detect its different functional effects.Results: ZNF382 was silenced due to promoter methylation in MKN45 and SGC7901 cell lines.In addition,the majority of tumor tissue samples had a lower level of ZNF382,while the adjacent non-tumor samples showed a higher ZNF382 level.Ectopic ZNF382 expression significantly inhibited gastric tumor cell clonogenicity,proliferation,migration and epithelial–mesenchymal transition(EMT);through inducing apoptosis.ZNF382 expression down-regulated the expression of SNAIL,Vimentin,Twist,NOTCH1,NOTCH3,NOTCH4,JAG1,MMP2,MMP11,as well as stem cell markers NANOG,OCT4 and SOX2.ZNF382 also up-regulated E-cadherin.Conclusions: To sum up,these results suggested that ZNF382 is downregulated in GC,and could act as a tumor suppressor gene via reverse the EMT in GC cells through NOTCH signaling.