NME2Reduces Proliferation, Migration And Invasion Of Gastric Cancer Cells To Limit Metastasis

Objective:(1) To detect the NME2expression of human gastric adenocarinoma specimens and analyze the relationship between NME2expression and various clinicopathological features.(2) To investigate the effect of NME2overexpression to cell growth, cell proliferation, cell migration and cell invasion in gastric adenocarinoma cells.(3) To explore preliminary the potential molecular mechanisms, of which NME2influence BGC823cells biological behavior.Methods:(1) Tissue chip and IM were used to measure the NME2expression of139cases with surgically removed gastric adenocarinoma tissues and analyze the relationship between NME2expression and gender, age, differentiation, primary tumor size, depth of invasion and lymph node metastasis.(2) Stable transfection was used to make two gastric adenocarinoma cell lines BGC823and MKN45overexpression NME2and indentify the NME2expression by RT-PCR and IF.(3) Flow cytometry detection, plat colony-formation assay, a wound-healing assay and Transwell chamber assay were used to observe the effect of incresed NME2expression to cell growth, cell proliferation, cell migration and cell invasion in gastric cancer cells.(4) RT-PCR was used to monitor the correlation between overexpression of NME2and expression of WNT2B, CD44and WISP-1to explore preliminary the potential molecular mechanisms.Results:(1) As shown in analysis of139cases with surgically removed gastric adenocarcinoma tissues, NME2expression was associated with differentiation and lymph node metastasis. There were a positive correlation between NME2expression and degree of differentiation (P<0.01), and the negative correlation between NME2expression and lymph node metastasis(P<0.05).(2) Stable transfection lead to the expression of NME2increased significantly in NME2-transfected cells when compared with that in Mock-transfeted cells and wild cells(P<0.05).(3) Overexpression of NME2had no effect on cell growth in BGC823and MKN45cells by flow cytometry detection (P>0.05).(4) Cell proliferation of BGC823cells and MKN45cells were limited in NME2-transfected cells when compared with that in Mock-transfeted cells and wild cells by plat colony-formation assay(P<0.05).(5) As a result of a wound-healing assay and Transwell chamber assay, cratches recovery time was delayed and the number of cells across the membrane was decresed in NME2-transfected cells when compared with that in Mock and wild cells(P<0.05).(6) The number of cells invaded to bottom chamber in NME2-transfected cells was less than that in Mock and wild cells by Transwell chamber assay (P<0.05).(7) Overexpression of NME2made the mRNA level of CD44and WISP-1decresed but the mRNA level of WNTB2in BGC823cells by RT-PCR.Conclusion:(1) NME2expression is associated with differentiation and lymph node metastasis of human gastric adenocarinoma. There are a positive correlation between NME2expression and degree of differentiation, and a negative correlation between NME2expression and lymph node metastasis. It suggests that NME2may promote differentiation and inhibit metastasis in the angiogenesis and progression of gastric adenocarinoma.(2)The result of increased NME2expression inhibiting cell proliferation, migration and invasion but the cell growth in BGC823and MKN45cells suggests that NME2may influence cell proliferation, cell migration and cell invasion of gastric adenocarcinoma cells to limit metastasis.(3) The potential molecular mechanisms, of which NME2influence BGC823cells biological behavior might be associated with the Wnt/β-catenin signaling pathway.