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Objective This study was to observe the biological effects of eprenone on proliferation ,migration and apoptosis in human gastric epithelial cell line .Methods Human gastric epithelial cells GES‐1 were cultured in vitro .MTT assay were used to e‐valuate the proliferation of GES‐1 cells in different concentrations of teprenone and ensure the appropriate drug concentration .T ran‐swell test and scratch test were used to detect the migration ability of GES‐1 cells treated with appropriate concentration of eprenone .Flow cytometry analysis were used to detect the apoptosis of GES‐1 cells treated by the appropriate concentration of eprenone .Results Treated with eprenone for 24 h ,the proliferation of GES‐1 cells were increased as the concentration of teprenone from 10 to 80 μmol/L ,but from 80 to 320 μmol/L ,the promoting effect showed no staticall significant changes .So the appropriate drug concentration was determined to be 80 μmol/L .Treated with teprenone (80 μmol/L) for 24 h ,the transwell test showed that the migration rate of the teprenone group was 3 .338 ± 0 .293 and the control group was 1 .328 ± 0 .208 .So the number of staining blue cells in eprenone group were more than in control group obviously under membrane of transwell chambers (P<0 .01) .Scratch test showed that the migration rate of the eprenone group was 1 .00 ± 0 .18 and the control group was 0 .72 ± 0 .08 .Similarly ,the migration rate of eprenone group was higher than the control group(P<0 .05) .Treated with teprenone (80 μmol/L) for 48 h ,the apoptosis rate of the teprenone group was (11 .90 ± 1 .53)% and the control group was (25 .61 ± 0 .15)% ,the cellular apoptosis of eprenone group was lower than the control group (P<0 .01) .Conclusion Teprenone can promote the proliferation and migration , inhibit the apoptosis of GES‐1 cells .
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Objective To determine the effect of different types of Helicobacter pylori(H, pylori) on the gap junction intercellular communication (GJIC) in GES-1 cells, and investigate the types of H. pylorirelated to the dysfunction of GJIC. Methods Different types of H. pylori clinical strains were isolated and cultured, including the East Asian CagA-positive H. pylori( East Asian CagA +H. pylorl), Western CagA-positive H. pylori( Western CagA +H. pylori), and the CagA-negative H. pylori (CagA-H. pylori). We co-cultured these H. pyloristrains with GES-1 cells for 24 and 48h, respectively. The control group was cultured without any H. pylorifor 24 and 48 h. Change of the GJIC function in GES-1 cells was detected by the scrape-loading dye transfer (SLDT) technique. The cell proliferation of each group was examined by the methyl thiazolyl tetrazolium bromide (MTT) assay. Results The control group showed better GJIC function in the GES-1 cells, and the fluorescent dye migrated 4 - 5 rows to the adjacent cells at 24 and 48 h. Compared with the control group, the GJIC function of GES-1 cells in the CagA - H. pylori group decreased and the fluorescent dye migrated 3 rows to the adjacent cells. Compared with the control group and the CagA- H. pylori group, the GJIC function of GES-1 cells in the Western CagA + H. pylori group decreased and the fluorescent dye migrated 1 - 2 rows to the adjacent cells. The East Asian CagA * H. pylori group showed no GJIC function or weak GJIC function, and most of the fluorescent dye was confined to the area of scratched single row cells and only a few migrated 1 -2 rows to the adjacent cells. Difference in the cell proliferation between the CagA - H. pylorigroup and the control group was not significant. The cell proliferation of the Western CagA + H. pylori group and the East Asian CagA + H. pylori group at bacterium-to-cell ratio of 100:1 and 200:1 was higher than that of the control group. The cell proliferation of the East Asian CagA +H. pylori group at bacterium-to-cell ratio of 400:1 was significantly lower than that of the control group at 48 h. Conclusion H. pylorican inhibit the GJIC function in GES-1 cells, which may be associated with CagA +H. pylori, especially with East Asian CagA +H. pylori. The effect of H. pylori on the proliferation of GES-I cells is related to virulence factor CagA.
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To observe the effect of Helicobacter pylori(H.priori) on cell gap junction ultrastructure of gastric epithelial cells,and to explore carcinogenic mechanism of H.priori from the changes of cell gap junction,BGC-823 cells were co-cultured with different H.prlori strains for 24 h and 48 h.The cell gap junction ultrastructure was observed under transmission electron microscope with sample preparation of fixation and embedding in situ.In 70 patients with gastric eancer(GC),H.priori was detected by rapid urease test,basic fuchsin stain and 14C-urea breath test.The CagA gene of H.prlori was determined by PCR and the cell gap junction ultrastructure was observed under transmission electron microscope.More cell gap junctions and junction complexes of BGC-823 cells were found in control group without H.priori.Groups with H.priori had less number of cell gap junctions,less number of junctions/unit perimeter,shorter length of junctions/unit perimeter,and bigger width of the intercellular space,comparing to control groups without H.priori(P＜ 0.001 or P＜ 0.005).The number of cell junctions and the number of junctions/unit perimeter in the groups co-cultured with NCTC J99,GC 01 and NCTC 11639(CagA+) were less than that in the groups co-cultured with NCTC 12908(CagA-)(P ＜0.001 or P＜0.05),and the length of junctions/unit perimeter in the groups co-cultured with NCTC J99 and GC 01 was shorter than that in the groups co-cultured with NCTC 12908 (P＜ 0.001).In patients with GC,the number of cell junctions,the number of junctions/unit perimeter and the length of junctions/unit perimeter in group H.priori infection were all less than those in group without H.prior/infection(P ＜0.001),and that in CagA+ H.prlori group were less than that in CagA- H.prlori group,but its smallest width of the intercellular space was longer than that in CagA- H.prlori group.The above results showed that the changes of cell gap junction of gastric epithelial cells were associated with H.prlori infection especially CagA+ H.prlori infection.
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PURPOSE: To investigate the relation of the gastric epithelial cell proliferation, apoptosis and genotypes of H. pylori in children. METHODS: Histologic grading by updated Sydney system, PCNA immunostaining, TUNEL method and the genotypes (cagA, picB and iceA) by PCR were performed in H. pylori positive (N=20) and negative (N=20) gastric biopsy specimens. RESULTS: PCNA index was significantly different between H. pylori positive children (77.4+/-13.12) and H. pylori negative children (52.3+/-12.20) (p=0.000). There were positive correlations between PCNA index and H. pylori density (r=0.624, p=0.000), polymorphonuclear neutrophil activity (r=0.460, p=0.005) and chronic inflammation (r=0.433, p=0.009). Apoptosis index of H. pylori positive children (0.70+/-0.411) was significantly higher than of H. pylori negative children (0.14+/-0.201) (p=0.000). Positive correlations between apoptosis index and H. pylori density (r=0.691, p=0.000), polymorphonuclear neutrophil activity (r=0.585, p=0.000) and chronic inflammation (r=0.535, p=0.001) were noted. As PCNA index increased, apoptosis index significantly increased (r=0.527, p=0.001). The positive rates of genotypes were cagA 90%, picB 75%, iceA1 60% and iceA2 15%, respectively. There were no significant correlations between the status of the genotypes and PCNA index, apoptosis index, the endoscopic findings and the histologic findings. CONCLUSION: PCNA index and apoptosis index in H. pylori positive children were higher than in H. pylori negative children but were not related to H. pylori genotypes. This study suggested that correlatively increased gastric epithelial cell proliferation and apoptosis are important to pathogenesis of H. pylori infection in children.
Subject(s)
Child , Humans , Apoptosis , Biopsy , Epithelial Cells , Genotype , Helicobacter pylori , Helicobacter , In Situ Nick-End Labeling , Inflammation , Neutrophils , Polymerase Chain Reaction , Proliferating Cell Nuclear AntigenABSTRACT
Objective To investigate the effects of sodium butyrate (SB) on fetal gastric epithelial cells (GECs) in vitro damaged by N-methyl-N’-nitro-N-nitrosoguanidine (MNNG). Methods The fetal gastric epithelial cells were isolated, cultured and identified. Then the cells of the second generation were cocultured with 10 -5 mol/L, 10 -6 mol/L, 10 -7 mol/L SB for 4 h, then added with 10 -5 mol/L MNNG culturing for another 24 h. After the treated cells were finally cultured in normal culture medium for 40 d, the unscheduled DNA synthesis (UDS), lipid peroxidation (LPO) and ras p21 were detected. The cells were only cultured with 10 -5 mol/L MNNG served as negative control and without MNNG or SB as normal control. Results The level of UDS, the contents of lipid peroxidation products and the concentrations of ras p21 in GECs treated with 10 -5 mol/L and 10 -6 mol/L SB decreased obviously, and had significant differences with that of GECs treated without SB. Conclusion SB can effectively prevent the damage of GECs induced from MNNG.
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PURPOSE: Dysregulation of gastric epithelial cell proliferation and apoptosis are important in development of ulcer, atrophy and neoplasia in Helicobacter pylori (H. pylori) infection. The aim of this study was to investigate the effect of infection of H. pylori on gastric epithelial cell proliferation and apoptosis in children. METHODS: Histological grading by updated Sydney system, PCNA immunostaining and TUNEL method were performed in H. pylori positive (N=58) and negative (N=40) gastric biopsy specimens. RESULTS: In H. pylori positive children, there were significantly higher grade of polymorphonuclear neutrophil activity (P=0.000), chronic inflammation (P=0.000), epithelial damage (P=0.000) and lymphoid follicles (P=0.000) than in H. pylori negative children. Intestinal metaplasia was not seen in H. pylori positive children. PCNA index was significantly different between H. pylori positive children (67.8+/-18.13) and H. pylori negative children (54.8+/-14.46, P=0.000). There was positive correlation between PCNA index and H. pylori density (r=0.277, P=0.007), polymorphonuclear neutrophil activity (r=0.280, P=0.007) and chronic inflammation (r=0.284, P=0.006). Apoptosis index of H. pylori positive children (0.44+/-0.447) was significantly higher than of H. pylori negative children (0.14+/-0.196, P=0.000). There was positive correlation between apoptosis index and H. pylori density (r=0.472, P=0.000), polymorphonuclear neutrophil activity (r=0.370, P=0.001) and chronic inflammation (r=0.483, P=0.000). There was positive correlation between PCNA index and apoptosis index (r=0.353, P=0.003). CONCLUSION: The PCNA and apoptosis index in H. pylori positive children were significantly higher than in H. pylori negative children. This study suggested that gastric epithelial cell proliferation and apoptosis are important to pathogenesis of H. pylori infection in children.
Subject(s)
Child , Humans , Apoptosis , Atrophy , Biopsy , Epithelial Cells , Helicobacter pylori , Helicobacter , In Situ Nick-End Labeling , Inflammation , Metaplasia , Neutrophils , Proliferating Cell Nuclear Antigen , UlcerABSTRACT
Objective To investigate the regulatory effects of bombesin on the growth of human immortalized gastric epithelial cell line(GES-1), and its mechanisms. Methods ① The expression of gastrin releasing peptide receptor(GRP-R) mRNA in GES-1 was detected. ② The expression of GRP-R protein was tested by cross-linking experiment and the location of the receptors in the cells were investigated by cytochemistry. ③GES-1 cell line was incubated with varying concentrations of bombesin with or without its antagonist and growth of the cell line was determined. ④The effect of protein kinase C (PKC) inhibitor on cell growth induced by bombesin was studied. ⑤After treated with bombesin, the intracellular IP 3 and translocation of PKC activity were measured in GES-1. ⑥Semiquantification of GRP-R mRNA in this cell line treated with bombesin was performed. Results ①Expression of mRNA of GRP-R was demonstrated in GES-1 cells. ②The GRP-R protein was about 75?103 as revealed by cross-linking study, and the receptors were identified on the cell membranes by cytochemistry. ③Bombesin stimulated the growth of GES-1 significantly, which could be inhibited by specific antagonist of bombesin. ④Bombesin-induced growth of GES-1 was also inhibited by PKC inhibitor. ⑤Bombesin induced an increase of IP 3 generation in GES-1 as well as remarkable translocation of PKC activity from cytoplasm to the cell membranes. ⑥An increase in GRP-R mRNA was induced by treatment of cell line with bombesin. Conclusions Bombesin stimulates the growth of this GES-1 via its receptor GRP-R and through IP 3, PKC signal pathway. The increase in expression of GRP-R mRNA in GES-1 induced by bombesin indicates that bombesin might upregulate the GRP-R in the GES-1 cells.