OBJECTIVETo establish two-dimensional electrophoresis profiles from human esophageal cancer tissue and paired normal esophageal tissue and identify differentially expressed proteins to identify the molecular markers for early-stage diagnosis.

METHODSThe total proteins of human esophageal cancer tissue and paired normal esophageal tissue were separated by immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The 6 differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The peptide mass fingerprintings (PMF) were identified by database searching. Six differentially expressed proteins were validated by RT-PCR.

RESULTSThe well-resolved, reproducible 2-DE patterns of esophageal cancer tissue and esophageal normal tissue were established. Using MALDI-TOF-MS technology, 6 differential protein spots were identified. Among them, the expressions of squamous cell carcinoma antigen 1 (SCCA1b), KRT4 and annexin A1 were downregulated and triosephosphate isomerase (TPI1), heat shock protein 27 (HSP27) and manganese superoxide dismutase (MnSOD) were upregulated in esophageal cancer tissues.

CONCLUSIONThe identification of differential expressed proteins in human esophageal cancer and normal tissue will be helpful for screening the biomarker for early-stage diagnosis.

Biomarkers, Tumor ; genetics ; metabolism ; Early Diagnosis ; Electrophoresis, Gel, Two-Dimensional ; Esophageal Neoplasms ; diagnosis ; genetics ; pathology ; Esophagus ; cytology ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Mass Spectrometry ; Neoplasm Staging ; Proteins ; genetics ; metabolism

Noncardiac chest pain (NCCP) is one of the most common esophageal symptoms and lacks a clearly defined mechanism. The most common cause of NCCP is gastroesophageal reflux disease (GERD). One of the accepted mechanisms of NCCP in a patient without GERD has been altered visceral sensitivity. Mast cells may play a role in visceral hypersensitivity in irritable bowel syndrome. In this case, a patient with NCCP and dysphagia who was unresponsive to proton pump inhibitor treatment had an increased esophageal mast cell infiltration and responded to 14 days of antihistamine and antileukotriene treatment. We suggest that there may be a relationship between esophageal symptoms such as NCCP and esophageal mast cell infiltration.
Adult ; Chest Pain/*etiology ; Esophageal Diseases/*complications/drug therapy ; Esophagus/cytology/pathology ; Female ; Histamine Antagonists/therapeutic use ; Humans ; Leukotriene Antagonists/therapeutic use ; Mast Cells/metabolism ; Mastocytosis/*complications/drug therapy

OBJECTIVETo explore the feasibility of detecting p53 gene mutation in exfoliative esophageal cells, and compare gene mutation between precancerous lesions and normal esophageal exfoliative cells and correlate p53 gene mutation with esophageal carcinogenesis.

METHODSForty-eight samples (24 normal squamous epithelia and 24 severe squamous dysplasia) were obtained by balloon cytologic technique from a high incidence area, Yanting county, Sichuan Province, China in 1982. p53 gene mutations in exons 5 and 7 were analyzed by PCR-SSCP.

RESULTSp53 genes were detected in all samples. Five samples with p53 mutation were detected in exon 7 and no mutation was detected in exon 5 in 24 severe dysplasia samples. Three of the 5 samples with mutation in exon 7 developed esophageal cancer in 1992, 1994 and 1996 respectively. No p53 gene mutation was detected in exon 5 and 7 in normal exfoliative samples.

CONCLUSIONp53 mutation may have occurred in the precancerous lesions which contributes in the initiation of human esophageal carcinogenesis.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Epithelial Cells ; pathology ; Esophageal Neoplasms ; genetics ; pathology ; Esophagus ; cytology ; pathology ; Exons ; genetics ; Female ; Follow-Up Studies ; Genes, p53 ; genetics ; Humans ; Male ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Precancerous Conditions ; genetics ; pathology

BACKGROUNDStudy on the promotive effects of N-nitrosopiperidine on carcinogenesis process was performed, based on the immortalization of human fetal esophageal epithelium induced by human papillomavirus (HPV) 18E6E7 genes.

METHODSThe immortalized esophageal epithelium SHEE was induced by HPV18E6E7. The cells at 17th passages were cultured in 50 ml flasks. The N-nitrosopiperidine (NPIP) 0, 2, 4, 8 mmol/L added to the cultured medium of SHEE cells for 3 weeks. The morphology, proliferation and apoptosis of the cells were studied by phase contrast microscopy and flow cytometry. Modal number of chromosomes was analyzed by standard method. Tumorigenicity of the cells was assessed by soft agar colony formation and by transplantation of cells into nude mice. Expression of HPV was detected by Western blot.

RESULTSWhen cells were exposed to high concentration (8 mmol/L) of NPIP, cell death was increased, leaving a few live cells. In normal cultural medium instead of NPIP proliferative status of the cells restored after 4 weeks and the cells progressed to the proliferation stage with continuous replication and atypical hyperplasia. At the end of the 8th week, the cells appeared with large colonies in soft-agar and tumor formation in transplanted nude mice. When the cells were cultured in 2, 4 mmol/L NPIP the doubling passage was delayed and without tumor formation in transplanted nude mice. Modal number of chromosomes was 61-65, in 8 mmol/L NPIP group and control group, 56-61. Expression of HPV18 appeared in experimental and control groups.

CONCLUSIONNPIP promotes malignant change of the immortalized esophageal epithelial cells induced by HPV18E6E7. HPV18E6E7 synergy with NPIP will accelerate malignant transformation in esophageal epithelium.

Animals ; Blotting, Western ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Cell Transformation, Viral ; drug effects ; DNA-Binding Proteins ; metabolism ; Epithelial Cells ; cytology ; drug effects ; virology ; Esophagus ; cytology ; Flow Cytometry ; Human papillomavirus 18 ; physiology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasms, Experimental ; metabolism ; pathology ; Nitrosamines ; toxicity ; Oncogene Proteins, Viral ; metabolism

OBJECTIVEImmortal cell line of human embryonic esophageal epithelium (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18 in our laboratory. To identify the fully malignant transformation at its 85th passage (SHEE85), the malignant phenotype, tumorigenesis and invasive potency were studied.

METHODThe cultured SHEE85 cells were observed under the light and the electron microscope (EM) for cell morphology, analyzed by flow cytometry for cell cycle. The tumorigenesis was assayed by plating cells in soft-agar and transplanting cells into the nude mice and SCID mice. To detect invasive potency, cells were cultured on amniotic membrane in vitro and transplanted into peritoneal cavity of mice in vivo.

RESULTSSHEE85 cells were crowded in cultivation with different sizes and shapes under light microscope, and displayed proliferative morphology under EM. Proliferative index was 47% with 12% hyperploidy cells in determination of DNA histogram. Many large colonies grew in soft-agar (4%) and the transplanted tumors were found in all 4 nude and 4 SCID mice, with strong invasive potency demonstrated in vitro and in vivo.

CONCLUSIONThe immortal esophageal epithelial cell line induced by HPV18 E6 E7 is derived from a fully malignant transformation with a strong invasive potency at the 85th passage. It is also a reliable model for studying the cellular and molecular mechanisms of carcinogenesis of the esophageal carcinoma.

Animals ; Cell Division ; genetics ; Cell Transformation, Neoplastic ; Cell Transformation, Viral ; genetics ; Cells, Cultured ; Epithelial Cells ; cytology ; ultrastructure ; virology ; Esophagus ; cytology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mice, SCID ; Microscopy, Electron ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Ploidies ; Transplantation, Heterologous

The anticarcinogenic effects and mechanisms of the biotechnological drugs of Panax ginseng C.A. Meyer cultivated in Russia, bioginseng, panaxel and panaxel- 5, were studied. Bioginseng was produced from a tissue culture of ginseng root cultured on standard medium, whereas panaxel and panaxel-5 were produced from ginseng tissue root cultures using standard mediums enriched with 2-carboxyethylgermanium sesquioxide and 1-hydroxygermatran-monohydrate respectively. All three ginseng drugs inhibited the development of mammary tumors induced by intramammary injections of N-methyl-N-nitrosourea (MNU) in rats, the development of the brain and spinal cord tumors induced by transplacental administration of N-ethyl-N-nitrosourea (ENU) in rats, and the development of uterine, cervical and vaginal tumors induced by intravaginal applications of 7,12-dimethylbenz(a)anthracene (DMBA) in mice. The ginseng drugs induced the cytotoxic activity of macrophages in mice, enhanced T-lymphocyte rosette formation in guinea pigs exposed to cyclophosphamide, and stimulated the production of thyroid hormones in rats. These mechanisms may contribute to the anticarcinogenic action of the ginseng drugs. The organic germanium compounds present in panaxel and panaxel-5 did not potentiate the anticarcinogenic or immuno- stimulatory effects as much as biogeinseng. Preliminary clinical trials with panaxel and bioginseng were carried out in patients with precancerous lesions of the esophagus and endometrium. Panaxel was found to have a strong therapeutic effect in patients suffering from chronic erosive esophagitis. Bioginseng induced the regression of adenomatous-cystic hyperplasia of the endometrium in some patients. Thus, we conclude that the drugs of ginseng appear to hold considerable promise for future cancer chemoprevention.
Adenocarcinoma/chemically induced/prevention & control ; Adult ; Animal ; Antineoplastic Agents, Phytogenic/*therapeutic use ; Cells, Cultured ; Cervix Neoplasms/chemically induced/prevention & control ; Clinical Trials ; Cytotoxicity Tests, Immunologic ; Disease Models, Animal ; Endometrial Neoplasms/pathology/prevention & control ; Endometrium/pathology ; Esophageal Neoplasms/pathology/prevention & control ; Esophagus/pathology ; Estradiol/blood ; Female ; Fibroadenoma/chemically induced/prevention & control ; Human ; Macrophages, Peritoneal/cytology/immunology ; Male ; Mammary Neoplasms, Experimental/chemically induced/prevention & control ; Mice ; Mice, Inbred C57BL ; Neoplasms, Experimental/chemically induced/*prevention & control ; Nervous System Neoplasms/chemically induced/prevention & control ; Panax/*metabolism ; Precancerous Conditions/pathology/*prevention & control ; Rats ; Tissue Culture ; Uterine Neoplasms/chemically induced/prevention & control ; Vaginal Neoplasms/chemically induced/prevention & control