OBJECTIVE: To investigate the expression of profilin 2 (PFN2) in gastric cancer and assess its potential value as a novel prognostic indicator and a therapeutic target. METHODS: We collected gastric cancer and paired adjacent tissues from 100 patients for immunohistochemical detection of PFN2 expression. According to the expression level of PFN2, the patients were divided into two groups with high (46 cases) and low (48 cases) PNF2 expression in cancer tissues, and also into two groups with high (26 cases) and low (49 cases) PNF2 expression in adjacent tissues. Chi-square test, Spearman correlation and KaplanMeier survival analysis were used to analyze the relationship between PFN2 protein expression level and the patients' clinical parameters. We also tested the effects of PFN2 knockdown and overexpression on the proliferation and migration of MKN-45 cells using Transwell assay and CCK-8 assay. RESULTS: The expression of PFN2 protein was significantly higher in gastric cancer tissues than in adjacent tissues (P < 0.01). PFN2 expression was positively correlated with M-stage of gastric cancer and VEGFR expression in the tumor tissues (P < 0.01). A high expression of PFN2 protein was significantly correlated with a poor prognosis of gastric cancer patients (P < 0.01), and was an independent predictor of the prognosis of gastric cancer. In MKN-45 cells, the cells overexpressing PFN2 showed significantly stronger proliferation and migration abilities than those with PFN2 knockdown (P < 0.001). CONCLUSION PFN2 protein is highly expressed in gastric cancer tissues to promote the proliferation and migration of the tumor cells. PFN2 may serve as a potential diagnostic marker, a prognostic indicator and a therapeutic target for gastric cancer.
Cell Proliferation ; Humans ; Profilins/metabolism* ; Prognosis ; Stomach Neoplasms/pathology* ; Survival Analysis

UNLABELLEDTo set up a new rapid and efficient way for the preparation of specific monoclonal antibodies (MAbs) with plasmid DNA immunization.

METHODSThe fusion gene of IL-2 signal peptide and profilin 1 by 'overlapping PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-IL2-profl. Another fusion gene of IgG kappa chain signal peptide and profilin 1 by 'no template PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-Igkappa-prof1. BALB/c mice were single intrasplenic immunized with plasmid DNA. Results After cell fusion and hybridomas screening by indirect ELISA, we charactered two mAbs (1D8A2B4 and 4B12A5E3) against profilin 1. The MAbs isotype were determined as IgM and IgG3.

CONCLUSIONA single intrasplenic plasmid DNA immunization is rapid and efficient and can be used as a helpful tool for the production of specific mAb.

Animals ; Antibodies, Monoclonal ; biosynthesis ; genetics ; immunology ; Antibody Specificity ; DNA ; genetics ; immunology ; Female ; Gene Fusion ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Profilins ; genetics ; immunology ; Spleen ; immunology ; metabolism