The Role And Mechanism Of NUP62 In Gastric Cancer

ObjectiveThe study was aim at detecting the expression of NUP62 in gastric cancer,adjacent non-cancerous tissues and various gastric cancer cell lines,exploring mechanism of NUP62 in the progress of gastric cancer,which will provide a new theoretical and research basis for the study of gastric cancer,and will supply a new diagnostic basis for molecular diagnosis and targeted therapy for gastric cancer.Methods1.The mRNA expression of NUP62 in gastric cancer tissues and normal tissues was investigated by GCBI(online database).Kaplan-Meier analysis was used to analyze the relationship between the expression of NUP62 in gastric cancer and the prognosis of patient.2.Immunohistochemical technique was used to detect the difference in expression of NUP62 in gastric cancer tissues and adjacent non-cancerous tissues.3.Western blot analysis was used to detect the difference in expression of NUP62 protein in several gastric cancer cell line.4.Using transfection technology to transfect overexpression plasmid(pcDNA-NUP62)into HGC-27 cells,Western blot to verify the transfection efficiency;RNA interference(RNAi)technology was used to inhibit NUP62 expression in SGC-7901 cells,and the knockdown effect was evaluated by Western blot.5.Colony formation assay,transwell migration experiment and wound healing assay was used to evaluate the proliferation and migration ability of gastric cancer cells;6.Western blot was used to detect the expression of proliferation,invasion and metastasis relative protein and expression of AKT,?-catenin and TGF-? pathways protein.Result1.Database analysis showed that the expression of NUP62 in gastric cancer tissues was significantly higher than that in normal tissues;Kaplan-Meier analysis showed that the OS and PFS of gastric cancer patients with high expression of NUP62 were decreased.2.The results of immunohistochemistry showed that NUP62 was highly expressed in the nucleus of gastric cancer cells and low in adjacent tissues.Western blot results showed that the expression of NUP62 in most gastric cancer cells was higher than that in normal gastric epithelial cells.The result agrees with the database result.3.The overexpression plasmid(pcDNA-NUP62)was transfected into HGC cells.The results of Western blot showed that NUP62 was overexpressed.The siRNA was transfected into SGC-7901 cells.The results of Western blot showed that NUP62 was successfully silenced.4.The results of plate colony formation assay,transwell migration experiment and wound healing assay showed that the cell proliferation and migration ability were significantly inhibited after silencing NUP62 in SGC-7901 cells.Further Western blot results showed that the expression of E-cadherin increased,the expression of N-cadherin and Snail decreased,the EMT occurrence was inhibited,the expression of Timp2 was increased,the expression of MMP2 and MMP9 was decreased,and the invasion and metastasis of gastric cancer cells were inhibited.While the expression of PCNA,c-myc and CyclinD1 was decreased,and the proliferation ability of gastric cancer cells was suppressed.After NUP62 was overexpressed in HGC-27 cells,the proliferation and migration ability increased significantly,the expression of E-cadherin decreased,the expression of N-cadherin and Snail increased,the expression of Timp1 decreased,the expression of MMP2 and MMP9 increased,and the invasion and metastasis of gastric cancer cells were promoted.The expression of PCNA,c-myc and CyclinD1 was increased,which promoted the growth of gastric cancer cells.5.In the PI3K/AKT pathway,after overexpressed NUP62 in HGC-27 cells,Western blot results showed that AKT and p-AKT protein expression was increased;when the NUP62 was silenced in SGC-7901 cells,AKT and p-AKT expression was declined.6.In the ?-catenin pathway,after overexpression of the NUP62 in HGC-27 cells,Western blot results showed that the expression of ?-catenin,GSK3? and p-GSK3? were increased;when NUP62 was silenced in SGC-7901 cells Western blot results showed that the expression of ?-catenin,c-myc,GSK3? and p-GSK3? were decreased.7.In the TGF-? pathway,when the expression of NUP62 was up-regulated in HGC-27 cells,the expression of TGF-?,SMAD2/3 and p-SMAD2/3 was increased by Western blot;while NUP62 was silenced in SGC-7901 cells.The expression of TGF-?,SMAD2/3 and p-SMAD2/3 was decreased.ConclusionsNUP62 is involved in the development and progression of gastric cancer,and induces the production of EMT in gastric cancer cells by regulating PI3K/AKT,?-catenin and TGF-? signaling pathways,and promotes the proliferation and migration of gastric cancer cells.NUP62 may become a new diagnostic indicator for clinical gastric cancer,providing a new theoretical basis for the diagnosis and treatment of gastric cancer.