【Abstract】ObjectiveTo investigate whether tumor necrosis factor-α (TNF-α) enhance the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9(MMP-9) in hepatic cancer cell line HepG2 or not. Methods Cultured HepG2 cells were treated by TNF-α with various concentration and time. The morphological changes of HepG2 cells were studied microscopically and the proliferation of HepG2 were detected by methyl thiazolyl tetrazolium (MTT). The expression of VEGF and MMP-9 mRNA in cultured HepG2 were determined by relative quantitative reverse transcription polymerase chain reaction. The VEGF and MMP-9 protein level in supernatants and in cytoplasm were determined by enzymelinked immunosorbent assay (ELISA) and by immunocytochemical staining, respectively.Results There was a little morphological changes in HepG2 with TNF-α treatment, but no change of cell proliferation in corresponding time. The expression of VEGF and MMP-9 mRNA was enhanced gradually with the TNF-α concentration increasing, the VEGF and MMP-9 protein level in supernatants and in cytoplasm was elevated gradually with the concentration increasing. There was a dependance on the concentration when the concentration of TNF-α was lower than or equal to 104 U/L. Furthermore, the effect of promotion was close to peak when the TNF-α concentration up to 104 U/L; but no timeeffect pattern observed. Conclusion TNF-αJP can enhance the expression of VEGF and MMP-9 at the level of mRNA and protein in hepatic cancer cell line.
Objective To observed the effect of IL-1β on expression of caudal-related homeobox gene 1 (CDX1) mRNA and mucoprotein 2 (MUC2) mRNA in cultured human gastric epithelial cells GES-1, and to investigate the underlying signal transduction pathways. Methods ①GES-1 cell was activated with IL-1β of different concentrations and time, the expression levels of CDX1 mRNA and MUC2 mRNA were detected by using real-time PCR. ②GES-1 cell was pretreated with PDTC, a NF-κB inhibitor, for 1 h prior to the addition of IL-1β, then the expressions of CDX1 mRNA and MUC2 mRNA were measured. Results Both CDX1 mRNA and MUC2 mRNA were not examined in GES-1 cell under normal culture conditions. But they could be induced by IL-1β with a dose-dependent manner in a concentration range (P＜0.05); 8 h after treatment with IL-1β, the peak values of the expression levels of CDX1 mRNA and MUC2 mRNA were reached (P＜0.05), then declined gradually. When pre-incubated with NF-κB inhibitor PDTC, the expression levels of CDX1 mRNA and MUC2 mRNA were significantly decreased (P＜0.05). Conclusion IL-1β significantly induces the expressions of CDX1 mRNA and MUC2 mRNA in cultured human gastric epithelial cell GES-1 through the NF-κB signal pathway, which indicates that IL-1β plays a role in the process of intestinal metaplasia.