OBJECTIVETo investigate the expression of neuron-specific enolase (NSE) and synaptophysin(SYN) proteins in different developmental stages of human embryonic esophagus.

METHODSImmunohistochemistry was used to detect the expressions of NSE and SYN proteins in embryonic esophagus tissues of fetuses of 2, 3 and 4 month gestational age (n=16). One-way ANOVA and LSD-t test were employed to compare the staining intensity and number of positive expression cells in embryonic esophageal tissues of different gestational age.

RESULTSIn fetuses with 2, 3 and 4 months of gestation, the number of NSE-positive nerve cells in the myenteric nerve plexus and submucosa of human embryonic esophageal tissues were 18.38 ± 8.37, 25.00 ± 11.54 and 38.00 ± 15.09, respectively; the staining intensity of NSE-positive nerve cells and nerve fibers in myenteric nerve plexus and submucosa of embryonic esophageal tissues were 74.38 ± 14.93, 62.25 ± 18.59 and 56.44 ± 14.70, respectively. NSE-positive cells were detected in the esophageal epithelium only at the third month. In the fetuses at 2, 3 and 4 months of gestation, SYN in all layers of esophageal tissue were positively or strong positively expressed, especially in the myenteric plexus and submucosal plexus. The staining intensity of SYN-positive cells in embryonic esophagus tissues of 2, 3 and 4 month gestation were 54.69 ± 9.34, 51.84 ± 6.10 and 46.41 ± 6.44, respectively.

CONCLUSIONSYN and NSE may be involved in the regulation of nerve system of esophageal tissues during the human embryonic development.

Esophagus ; embryology ; Female ; Fetus ; Gestational Age ; Humans ; Immunohistochemistry ; Phosphopyruvate Hydratase ; metabolism ; Pregnancy ; Synaptophysin ; metabolism

OBJECTIVEThe conversion of arginine into citrulline, termed citrullination, has important consequences for the structure and function of proteins. The present study aimed to identify novel citrullinated proteins in 10 tumor cell lines by 2-D Western blotting (2-D WB).

METHODSTwo identical two-dimensional electrophoresis (2-DE) gels were prepared using extracts from ten cultured human tumor cell lines: ECA(esophageal cancer cells), HEPG2 (hepatocellular carcinoma cells), SKOV3 (ovarian cancer cells), MCF-7 (breast cancer cells), H292 (lung mucoepidermoid carcinoma cells), HeLa (cervical cancer cells), Lovo (colon cancer cells), OS-RC (renal cell carcinoma cells), PANC-1 (pancreatic cancer cells), and SGC (gastric cancer cells). The expression profiles on one 2-DE gels were trans-blotted to PVDF membranes, and the blots were then probed with an anti-citrulline antibody. By comparing the 2-DE profile with the parallel 2-D WB profile at a global level, protein spots with immuno-signals were collected from the second 2-DE gel and identified using mass spectrometry. Immunoprecipitation was used to verify the expression and citrullination of the targeted proteins in the tumor cell lines.

RESULTS2-D WB and mass spectrometry identified citrullinated ENO1 (α-enolase), HSP60 (heat shock protein 60), KRT8 (keratin 8), TUBB (tubulin beta), TCRβ (T cell receptor β chain), VIME (vimentin) and PDI in these cell lines. Immunoprecipitation analyses verified the expression and citrullination of ENO1, HSP60, KRT8, and TUBB in the total protein lysates of the tumor cell lines.

CONCLUSIONThe citrullination of proteins ENO1, HSP60, KRT8, and TUBB suggests a new mechanism in the tumorigenic process.

Blotting, Western ; Cell Line, Tumor ; Citrulline ; metabolism ; Female ; Humans ; Immunoprecipitation ; Mass Spectrometry ; Phosphopyruvate Hydratase ; Vimentin

Carcinoid Tumor ; metabolism ; pathology ; surgery ; Female ; Humans ; Middle Aged ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism ; Thymectomy ; Thymus Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo investigate the distribution patterns of neuron-specific enolase (NSE) and synaptophysin (SYN) during the development of human fetal stomach.

METHODSSixteen specimens of human fetal (gestational age 2 to 4 months) gastric tissues were examined with immunohistochemistry for detecting the distribution of NSE and SYN expressions in the gastric walls.

RESULTSDuring the second to fourth gestational months, NSE was strongly expressed in the nerve cells and nerve fibers of the myenteric nerve plexus of human fetal stomach. As the gestational age increased, the numbers of NSE positive cells and fibers increased gradually in the gastric submucosa, but NSE was negative in the gastric mucosa. At the second gestational month, SYN expression was negative in the mucosa but positive in the myenteric nerve plexus; during the third to fourth months, positive SYN expression was found in the mucosa, submucosa and myenteric nerve plexus of the embryonic gastric walls and its expression intensity increased with the gestational age.

CONCLUSIONSYN and NSE are both involved in the regulation of the nervous system in the gastric wall but their expressions and distributions follow different patterns during the development of human fetal stomach.

Fetus ; metabolism ; Gastric Mucosa ; metabolism ; Gestational Age ; Humans ; Immunohistochemistry ; Myenteric Plexus ; Nerve Fibers ; metabolism ; Neurons ; metabolism ; Phosphopyruvate Hydratase ; metabolism ; Stomach ; metabolism ; Synaptophysin ; metabolism

Adenocarcinoma ; metabolism ; pathology ; Aged ; Carcinoid Tumor ; metabolism ; pathology ; Chromogranin A ; metabolism ; Humans ; Immunohistochemistry ; Male ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Rectal Neoplasms ; metabolism ; pathology ; S100 Proteins ; metabolism

PURPOSE: This study tried to identify novel gastric autoimmune antigens that might be involved in aggravating the atrophic gastritis among patients with Helicobacter pylori infection using two-dimensional immunoblotting analysis. MATERIALS AND METHODS: Proteins from gastric mucosal antrectomy specimens and AGS cells (gastric adenocarcinoma cell lines derived from a Caucasian patient who had received no prior therapy) were 2-dimensionally immunoblotted separately with a pool of 300 sera from H. pylroi-infected patients at Gyeongsang National University Hospital. RESULTS: Thirty-eight autoantigenic proteins including alcohol dehydrogenase [NADP+], alpha enolase, gastrokine-1, gastric triacylglycerol lipase, heat shock 70 kDa protein 1, and peroxiredoxin-2 were identified in the gastric mucosal tissue. Fourteen autoantigenic proteins including programmed cell death 6-interacting protein, serum albumin and T-complex protein 1 subunit gamma were identified in the AGS cells. Albumin, alpha-enolase, annexin A3, cytoplasmic actin 1, heat shock cognate 71 kDa protein and leukocyte elastase inhibitor were commonly observed autoantigenic proteins in both gastric mucosal tissue and AGS cells. Alpha-enolase, glutathione S-transferase P, heat shock cognate 71 kDa protein, heat shock 70 kDa protein 1, human mitochondrial adenosine triphosphate synthase (ATP) subunit beta, mitochondrial 60 kDa heat shock protein, peroxiredoxin-2, 78 kDa glucose-regulated protein precursor, tyrosine-protein phosphatase non-receptor type 11 and Tryptophan-Aspartic acid (WD) repeat-containing protein 1 showed 60% or higher amino acid positivity. CONCLUSION: These newly identified gastric autoimmune antigens might be useful in the control and prevention of gastroduodenal disorders, and might be valuable in breaking the vicious circle that exists in gastroduodenal disorders if their pathophysiological roles could be understood in the progress of chronic atrophic gastritis, gastroduodenal ulcers, intestinal metaplasia, and gastric carcinogenesis.
Alcohol Dehydrogenase/metabolism ; Autoantigens/*metabolism ; Electrophoresis, Gel, Two-Dimensional ; Gastric Mucosa/metabolism/microbiology ; Helicobacter Infections/*metabolism ; Humans ; Peptide Hormones/metabolism ; Phosphopyruvate Hydratase/metabolism

Antigens, CD34 ; metabolism ; Cytokines ; metabolism ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Perineum ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Sarcoma ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Adult ; Chromogranin A ; metabolism ; Female ; Humans ; Immunohistochemistry ; Pancreatic Neoplasms ; diagnosis ; metabolism ; therapy ; Phosphopyruvate Hydratase ; metabolism ; Somatostatin ; metabolism ; Somatostatinoma ; diagnosis ; metabolism ; therapy ; Synaptophysin ; metabolism ; Tomography, X-Ray Computed

Aged ; Back ; pathology ; CD57 Antigens ; metabolism ; Chondrosarcoma ; metabolism ; pathology ; Diagnosis, Differential ; Fibroma, Ossifying ; metabolism ; pathology ; Humans ; Male ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism ; Soft Tissue Neoplasms ; metabolism ; pathology

Adult ; CD56 Antigen ; metabolism ; Carcinoma, Neuroendocrine ; metabolism ; pathology ; surgery ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Mediastinal Neoplasms ; metabolism ; pathology ; surgery ; Mediastinum ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism