The CCDC43-ADRM1 Axis Regulated By YY1,Promotes Proliferation,invasion And Metastasis Of Gastric Cancer

Backgroud and ObjectionGastric cancer(GC)is the second leading cause of cancer-related death worldwide.Most patients are detected with metastasis,due to the lack of symptoms in the early stages of GC.The 5-year survival rate of patients with advanced GC is low for the limited treatment.Therefore,the mechanism of metastasis and growth of GC needs to be further studied.In recent years,studies have shown that coiled coil domain-containing(CCDC)gene is closely associated with many malignant tumor proliferation and metastasis.Coiled-coil domain containing 43(CCDC43)gene is located on chromosome 17,and CCDC43 gene is highly conserved in humans,rhesus monkeys,mice,rats,dogs,and zebrafish.The Gene Expression Profiling Interactive Analysis(GEPIA)database shows that CCDC43 mRNA is up-regulated in ovarian,lung,head and neck,cervical,breast,adrenal,stomach,liver and colon cancers.Our previous results confirmed that CCDC43 is highly expressed in colorectal cancer and promotes colorectal cancer metastasis by mediating epithelial-mesenchymal transition(EMT).However,the biological function of CCDC43 in gastric cancer is still not clear.ADRM1 is determined as a potential downstream target of CCDC43 by isobaric tags for relative and absolute quantitation(iTRAQ),and CCDC43 interacts with ADRM1 in GC cells.Bioinformatic analysis indicates that YY1 is the co-transcription factor of CCDC43 and ADRM1.This study mainly explores the mechanism of CCDC43 in proliferation and metastasis of GC and provides certain experimental basis for the targeted therapy of GC.MethodsFirstly,the expression level of CCDC43 in GC tissues was detected by real-time quantitative PCR and immunohistochemical(IHC)staining.The relationship between CCDC43 expression and clinicopathological parameters and prognosis was also analyzed.GC cells were transfected with CCDC43 plasmid or CCDC43 siRNA to detect the effect of CCDC43 on GC cells proliferation,invasion and migration.Finally,western blot was used to detect the effect of CCDC43 on EMT.iTRAQ were then used to screen ADRM1 as a potential downstream target of CCDC43.We used co-immunoprecipitation to verify whether CCDC43 and ADRM1 interact in GC cells.Immunofluorescence was performed to detect subcellular localization of CCDC43 and ADRM1.CHX was added to detect the influence of CCDC43 on the stability of ADRM1.We used a series of experiments including in vitro and in vivo to analyze the effect of CCDC43 on the proliferation,invasion and metastasis of gastric cancer via ADRM1.Biological information analysis YY1 is the co-transcription factor of CCDC43 and ADRM1.The binding relationship between YY1 and CCDC43 or ADRM1 promoter was analyzed by chromatin immunoprecipitation assay(ChIP)assay and luciferase reporter assay.The effect of YYlon CCDC43-ADRM1 axis on the biological function of GC cells was determined by a series of in vitro experiments.Finally,the correlation of YY1/CCDC43/ADRM1 in GC tissues was analyzed by western blot and IHC.Results1.CCDC43 is highly expressed in GC tissues,and the expression level of CCDC43 is related to the prognosis of patients.CCDC43 can promote the proliferation,invasion and metastasis of GC cells,and the expression of CCDC43 is related to the tumor differentiation,lymph node metastasis,AJCC staging,tumor invasion,and tumor size.2.CCDC43 interacts with ADRM1 in the cytoplasm.CCDC43 can stabilize ADRM1.CCDC43 promotes proliferation,invasion and metastasis of gastric cancer cells via ADRM1.3.YY1 is a co-transcription factor of CCDC43 and ADRM1.CCDC43-ADRM1 axis,regulated by YY1,promotes proliferation and metastasis of GC cells.YY1,CCDC43 and ADRMI were positively correlated in GC tissues.ConclusionsCCDC43 is highly expressed in gastric cancer tissues.CCDC43-ADRM1 axis,regulated by YY1,promotes proliferation,invasion and metastasis of gastric cancer...