Expression Of Prolyl Hydroxylase 3 In Gastric Cancer And It's Effects On Clone Formation And Migration Of Gastric Cancer Cells

Objective:To investigate the expression of Prolyl hydroxylase-3(PHD3)in gastric cancer,the effects of PHD3 on colony formation and migration of gastric cancer cells and potential molecular mechanism.Methods:Immunohistochemistry and Western-blot are used to explore the relationship between expression of PHD3 and pathological characteristics of neoplasm in 60 gastric cancer tissues.Cell transfection and RNA Interference(RNAi)are used to acquire AGS?BGC823 gastric cancer cell lines which over-expressed and silenced PHD3.In these cell lines,soft agar assay and chamber assay are used to test the changes.We use immunofluorescence,immunoprecipitation and Western-blot to test the interaction of PHD3 and Dishevelled-2(DVL2)in AGS and BGC823 cell lines,and the expression of Cyclin D1,c-Myc and Twist in different groups,which were the downstream target genes of Wnt/?-catenin/TCF signaling pathway.Results:The expression of PHD3 declines in gastric cancer tissues comparing with para-carcinoma tissue(P<0.01).The expression of PHD3 is related with the size of tumor.PHD3 expresses lower in bigger tumor(P<0.05).The level of expression affect by the composition of different cancer stage(P<0.05).The expression of PHD3 declines in advanced stage tumor by Wilcoxon rank sum test(P<0.01).The differences of PHD3 expression have no obvious statistical significance in disparate gender,age,the location and pathologic types of tumor.The expression of PHD3 rises after shifting into PHD3 expression vector and declines after shifting into PHD3 shRNA lentiviral vector in AGS and BGC823 gastric cancer cell lines by Western-blot.Clone formation and migration fall off in over-expressed PHD3 cells(P<0.01)and go up in low-expressed PHD3 cells(P<0.01)in AGS and BGC823 gastric cancer cell lines.The expression of Cyclin D1,c-Myc and Twist down-regulate in over-expressed PHD3 cells and up-regulate in low-expressed PHD3 cells.There is interactionbetween PHD3 and DVL2 in gastric cancer cells.Conclusion:PHD3 may be a tumor suppressor gene and low expressed in gastric cancer.It can interact with DVL2 to restrain Wnt/?-catenin/TCF signal pathway to reduce proliferation,invasion and metastasis of gastric cancer cells.This may be a new target spot to cure gastric cancer.