The Function And Mechanism Of ZNF382 On Cell Growth And Metastasis In Esophageal Squamous Cell Carcinoma

Objective: Esophageal carcinoma which occurred in the esophageal mucosa epithelium is the nineth most common cancer in the worldwide.There are approximately 300 thousand people die of esophageal cancer annually.According to the estimated data from World Cancer Report,the number of cancer related deaths of esophageal cancer ranks in the seventh among all cancer cases.China contributed to the highest incidence mortality rate of esophageal cancer.The estimated number of cancer cases and cancer deaths among men are much higher than women.Although the incidence of ESCC has declined,5-year survival rate still remains poor.Esophageal cancer included two types.Esophageal squamous cell carcinoma(ESCC)which comprises 90% of esophageal cancer is the predominant one in China,and the remainder is esophageal adenocarcinoma(EAC).The incidence of esophageal cancer varies geographically.Presently,tobacco and alcohol consumptions are recognized as the major risk factors of ESCC.More and more researches have reported that genetic mutations,epigenetic and other environmentalfactors can also induce esophageal cancer.Landscape of genetic alterations across the 144 Japanese ESCC samples evaluated the association of genetic alterations with patient prognosis and mutational signatures,fully illustrated that gene mutation and molecular genetic alterations are closely related to the occurrence and development of ESCC.Zinc finger proteins(ZFPs)are the largest transcription factor family in mammals.About one-third of ZFPs are Krüppel-associated box domain(KRAB)-ZFPs and regulate cell differentiation/proliferation/apoptosis and neoplastic transformation by sequence specific binding to DNA.We recently identified ZNF382 as a newly 19q13.12 tumor suppressor gene,epigenetically inactivated in carcinomas due to frequent promoter CpG methylation.However its methylation status,biological functions/mechanism and clinical significance in esophageal squamous cell carcinoma(ESCC)are still obscure.Here we mainly investigated the function and mechanism of ZNF382 on esophageal squamous cell carcinoma.Methods and Results: We characterized ZNF382 expression in ESCC and paired adjacent non-cancer tissues by qRT-PCR and investigated ZNF382 promoter methylation status in ESCC by MSP.We showed that ZNF382 expression was suppressed in ESCC due to aberrant promoter methylation,but highly expressed in normal esophagus tissues.Then we transfected ZNF382 plasmid in KYSE150,KYSE410 and KYSE510 cellsand knockdown the endogenous expression of ZNF382 in EC1 cell for cell functional experiments.Restoration of ZNF382 expression in silenced ESCC cells suppressed cell proliferation and metastasis,and induced tumor cell apoptosis.To further investigate the subtle mechanism of ZNF382 in ESCC,RNA-Sequence was performed to identify differentially expressed genes regulated by ZNF382.Online CHIP-Sequence data and TCF/LEF dependent luciferase reporter assay further analyzed the effect on Wnt/?-catenin signaling by ZNF382 in ESCC,and found ZNF382 suppressed Wnt/?-catenin signaling and the downstream target gene expression likely through directly binding to FZD1 and DVL2 promoters.Conclusion: In summary,our findings demonstrate that ZNF382 acts as a bona fide tumor suppressor,suppresses Wnt/?-catenin signaling pathway through directly binding to FZD1 and DVL2 genes promoters,inhibiting ESCC pathogenesis.