Animals ; Antigens, Neoplasm ; biosynthesis ; genetics ; immunology ; Apoptosis ; physiology ; Breast Neoplasms ; immunology ; metabolism ; CD3 Complex ; immunology ; Female ; Humans ; Killer Cells, Natural ; pathology ; Neoplasms ; immunology ; metabolism ; Receptors, Antigen, T-Cell ; immunology ; Stomach Neoplasms ; immunology ; metabolism ; Uterine Cervical Neoplasms ; immunology ; metabolism

BACKGROUND: It is well known that both gastric and intestinal phenotypic cell markers are expressed in gastric cancers. This study was aimed at investigating the correlation between gastric and intestinal phenotypic marker expression patterns of tumors and the clinicopathologic characteristics of gastric carcinomas. METHODS: We evaluated phenotypic marker expression by immunohistochemical staining with monoclonal antibodies. All tumors were classified as gastric (G), gastric and intestinal mixed (GI), intestinal (I), or null (N) phenotype. RESULTS: The tumors were phenotypically divided into G-phenotype tumors (33.2%), GI-phenotype tumors (25.7%), I-phenotype tumors (26.8%), and N-phenotype tumors (14.3%). N-phenotype tumors were associated with more corporeal location than GI- and I-phenotype tumors (p=0.009 and p=0.007, respectively), a larger size than I-phenotype tumors (p=0.007), a higher proportion of advanced gastric cancers than G-, GI-, and I-phenotype tumors (p=0.003, p< 0.001, and p< 0.001, respectively), more perineural invasion than G-, GI-, and I-phenotype tumors (p=0.076, p=0.003, and p=0.003, respectively), and more lymph node metastasis than GI-phenotype tumors (p=0.017). I-phenotype tumors were associated with a higher proportion of intestinal-type tumors than G-, GI-, and N-phenotype tumors (p< 0.001, p=0.011, and p< 0.001, respectively). CONCLUSION: Our results indicate that the gastric and intestinal phenotypic marker expression pattern of tumors is prognostically useful for patients with gastric carcinoma.
Tumor Markers, Biological/genetics ; Stomach Neoplasms/genetics/immunology/*pathology ; Prognosis ; *Phenotype ; Middle Aged ; Male ; Intestinal Neoplasms/genetics/immunology/*pathology ; Immunohistochemistry ; Humans ; Female ; Carcinoma/genetics/immunology/*pathology ; Antibodies, Neoplasm ; Antibodies, Monoclonal

CD99 is characteristically expressed in Ewing's sarcoma/primitive neuroectodermal tumor. Recently its immunoreactivity has also been reported in other tumors. However, the significance of CD99 isoforms expressed in these tumors has not been elucidated. In this study, we evaluated the expression of CD99 isoforms and its relationship with histopathologic parameters in gastric adenocarcinomas. Paraffin sections of 46 gastric adenocarcinomas were stained with an anti-CD99 monoclonal antibody, YG32. Twelve (26.1%) cases of 46 gastric adenocarcinomas showed immunoreactivity to YG32. The CD99 expression was also seen both in non-neoplastic foveolar epithelial cells and infiltrating lymphocytes. In addition, Western blot and RT-PCR analyses revealed that the type I is the predominant isoform of CD99 in non-neoplastic and neoplastic gastric tissues. The CD99 expression was usually seen in the intestinal type adenocarcinoma, while rarely in the diffuse type. The CD99 immunoreactivity decreased in MMP-2-overexpressing adenocarcinomas (p=0.028). Our results suggest that the type I is the major isoform of CD99 expressed in non-neoplastic gastric mucosa and gastric adenocarcinomas and its downregulation in gastric adenocarcinoma may be associated with cellular dedifferentiation and/or MMP-2 overexpression.
Adenocarcinoma/*immunology/pathology ; Adult ; Aged ; Antigens, CD/*analysis/genetics ; Cell Adhesion Molecules/*analysis/genetics ; Female ; Gastric Mucosa/cytology/immunology ; Humans ; Male ; Matrix Metalloproteinases/metabolism ; Middle Aged ; Protein Isoforms/analysis/genetics ; RNA, Messenger/genetics/metabolism ; Stomach Neoplasms/*immunology/pathology

To investigate the effects of dendritic cells (DCs) transfected with full length wild type p53 and modified by gastric cancer lysates on immune response, the wild type P53 was transducted to DCs with adenovirus, and the DCs were modified by gastric cancer lysates (Lywt-P53DC). The concentration of the surface molecules (B7-1, B7-2, MHC-I , MHC-II) of all DCs was determined by FACS, and the ability of the DCs to induce efficient and specific immunological response in anti-51Cr-labeled target cells studied. BALB/c mice model infected with DCs and Mk28 was established. CTL response in mice immunized with Lywt-p53DC and the effectiveness of Lywt-p53DC in the treatment of tumor-bearing mice was assayed. FACS revealed that the surface molecules of Ly-wt-P53 DC had a high expression: for B7-1 86.70% +/- 0.07%, B7-2 18.77% +/- 0.08%, MHC-I 87.20% +/- 0.05%, MHC-II 56.70%+/-0.07%; The T lymphocytes had a specific CTL lysing ability induced by Lywt-P53DC with the CTL lysis rate being 81%. The immune protective effect of Lywt-p53DC group was more obvious than any other groups (P<0.05). The tumor diameter in Lywt-p53DC group was 3.10+/-0.31 mm, 2.73+/-0.23 mm, 3.70+/-0.07 mm on the day 13, 16 and 19, smaller than DC, wtp53DC and LyDC groups (P<0.05). On the other hand, the growth rate of tumor in Lywt-p53DC group was slower than any other groups (P<0.05). It was suggested that DCs transfected with wild type P53 and modified by gastric cancer lysates had specific CTL killing capability.
Adenoviridae ; genetics ; metabolism ; Animals ; Cell Line, Tumor ; Dendritic Cells ; immunology ; metabolism ; Female ; Genetic Vectors ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; genetics ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of gastrin in human gastric cancer cell line SGC-7901 and the effects of gastrin-17 and anti-gastrin mAb on its growth.

METHODSThe expression of gastrin was determined by immunohistochemistry with anti-gastrin mAb prepared by our group. In a series of experiments, the growth of SGC-7901 cells was evaluated by MTT assay on cells grown in serum-free medium and treated with gastrin-17 and/or anti-gastrin mAb.

RESULTSImmunohistochemical examination of SGC-7901 cells revealed a specific gastrin immunoreactivity. Gastrin-17 significantly stimulated cell growth at the concentrations of 1 x 10(-9) mol/L approximately 1 x 10(-5) mol/L in a dose-dependent manner. The growth of SGC-7901 cells treated with anti-gastrin mAb, either alone or in combination with gastrin-17 (1 x 10(-7) mol/L), was significantly inhibited.

CONCLUSIONGrowth of human gastric cancer cells SGC-7901 can be stimulated in an autocrine fashion. The gastrin-stimulated growth of gastric cancer cells can be blocked by anti-gastrin mAb bound specifically with gastrin. Further study on the significance of anti-gastrin mAb in designing immunotherapy targeting to gastrin or gastrin receptor is warranted.

Antibodies, Monoclonal ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Gastrins ; biosynthesis ; genetics ; immunology ; Humans ; Receptor, Cholecystokinin B ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the relationship between the expression of host human leukocyte antigen-B mRNA (HLA-B mRNA) and HLA-B antigen in peripheral blood leukocytes (PBLs) and the differentiation and metastasis of gastric carcinoma (GC).

METHODSTo design and screen specific primers of HLA-B gene independently, detect the expression of HLA-B mRNA from 30 GC patients by reverse transcription-PCR and compare with the HLA-B antigen expression measured by flow cytometry.

RESULTSThe expression rate of PBL HLA-B mRNA from GC patients (23. 3% ) was very significantly lower than that of normals (87. 5% ) (P <0. 01) , especially concerning the poorly differentiated GC patients with lymph node metastasis (16. 0% ). Measured by flow cytometry, the expression percentage of HLA-B antigen of well-differentiated GC patients without lymph node metastasis was 88. 2% , an obviously decreasing tendency was showed in comparison with that in the normal group (98. 8% ) , although the difference was not significant (P = 0. 056) , and the expression percentage in poorly differentiated GC patients with lymph node metastasis(73. 3% )was declined significantly (P <0. 05).

CONCLUSIONThe expression of PBL HLA-B mRNA and HLA-B antigen in GC patients is decreased or lost, and correlated with differentiation and metastasis of the cancer. The expression of PBL HLA-B mRNA may more directly reflect its relationship with the tumor differentiation and metastasis than that of HLA-B antigen.

Adult ; Aged ; Cell Differentiation ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; HLA-B Antigens ; analysis ; genetics ; Humans ; Leukocytes ; immunology ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; immunology ; pathology

Inflammation is closely related to the progression of cancer as well as tumorigenesis. Here, we investigated the effect of prostaglandin E2 (PGE2) and interleukin-1beta (IL-1beta) on E-cadherin expression in SNU719 gastric cancer cells. E-cadherin expression decreased as the dose or exposure time of PGE2 and IL-1beta increased, whereas Snail expression increased with dose or time of PGE2 and IL-1beta. E-cadherin expression reduced by PGE2 treatment increased after the transfection of Snail siRNA. Neutralization of IL-1beta using anti-IL-1beta antibody blocked the expression pattern of E-cadherin and Snail occurred by IL-1beta treatment. However, there was no synergic effect of IL-1beta and PGE2 on the expression pattern of E-cadherin and Snail. In conclusion, inflammatory mediators reduced E-cadherin expression by enhancing Snail expression in gastric cancer cells. Inflammation-induced transcriptional regulation of E-cadherin in gastric cancer has implications for targeted chemoprevention and therapy.
Antibodies/immunology ; Antineoplastic Agents/pharmacology ; Cadherins/*metabolism ; Cell Line, Tumor ; Dinoprostone/*pharmacology ; Gene Expression Regulation/*drug effects ; Humans ; Interleukin-1beta/immunology/*pharmacology ; RNA Interference ; RNA, Small Interfering/metabolism ; Stomach Neoplasms/metabolism/pathology ; Transcription Factors/antagonists & inhibitors/genetics/*metabolism

OBJECTIVEThe B7-H1/PD-1 co-signaling pathway has recently been found to play a pivotal role in the immune evasion of tumor cells from host immune system. The aim of this study was to examine the B7-H1 and PD-1 expression and TILs status in gastric cancer and to elucidate the clinical relevance of B7-H1 and PD-1 to the pathogenesis of gastric carcinoma.

METHODSImmunohistochemistry and ANAE histochemical staining were used to investigate the in situ expression of B7-H1 and PD-1 and TILs status in the gastric tissues. RT-PCR was used to explore B7-H1 and PD-1 expression at the transcriptional level. The B7-H1 expression at protein level was detected by Western blot.

RESULTSExpression of B7-H1 and PD-1 was found to be increased in gastric carcinoma, but absent in normal gastric tissue. B7-H1 expression in gastric carcinoma was inversely correlated with TILs infiltration. B7-H1 but not PD-1 expression in tumor tissue was significantly correlated with some clinicopathhological variables including depth of invasion, lymph node metastasis and distant metastasis.

CONCLUSIONB7-H1 and PD-1 expressions are increased in gastric carcinoma. This signaling pathway may inhibit antitumor immune responses in gastric carcinoma. B7-H1 expression plays a critical role in the pathogenesis of human gastric carcinoma,and might be a promising prognostic marker and therapeutic target in the treatment of this disease.

Adult ; Aged ; Antigens, CD ; genetics ; metabolism ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; B7-H1 Antigen ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Lymphatic Metastasis ; Lymphocyte Subsets ; immunology ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Programmed Cell Death 1 Receptor ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; genetics ; immunology ; pathology

OBJECTIVETo study the characteristics of host immune response against human gastric carcinoma of different histological types.

METHODSThe expression of 24 families of T cell receptor beta chain variable region (TCRVbeta) and cytokine profiles in isolated CD4(+) and CD8(+) subsets, as well as the cytokine profiles in purified epithelial cells from the tumor tissue and the residual benign tissue of patients with gastric carcinoma, was detected by a highly sensitive radioactivity labeled semi-quantitative RT-PCR technique.

RESULTSThe number of expanded T cell clones in CD8(+) subset from tumor tissue of the intestinal-type carcinoma was larger than that of diffuse-type (P = 0.046). The mRNA levels of IL-6, IL-8 in CD8(+) T subset, as well as the level of TNF-alpha in CD4(+) T subset from the tumor tissue of the diffuse type (0.61 +/- 0.29, 0.56 +/- 0.22, 0.09 +/- 0.03) were significantly higher than that from the residual benign tissue (0.14 +/- 0.05, 0.27 +/- 0.09, 0.04 +/- 0.02; P = 0.028, P = 0.043, P = 0.046). However, the mRNA level of IL-8 in CD8(+) subset and epithelial tumor cells of the intestinal-type (0.57 +/- 0.25, 0.27 +/- 0.07) was significantly higher than that from the residual benign tissue (0.21 +/- 0.07, 0.14 +/- 0.06; P = 0.028, P = 0.028).

CONCLUSIONSThe characteristics of host immune response against tumor are different between intestinal-type and diffuse-type gastric carcinoma. Both fewer expanded T cell clones and more suppressive cytokines suggest a more suppressive immune status in the local tumor lesion of diffuse-type than in the intestinal-type of the gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; CD4-Positive T-Lymphocytes ; metabolism ; pathology ; CD8-Positive T-Lymphocytes ; metabolism ; pathology ; Clone Cells ; Cytokines ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Interleukin-2 ; genetics ; Interleukin-4 ; genetics ; Interleukin-6 ; genetics ; Interleukin-8 ; genetics ; Interleukins ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; Stomach Neoplasms ; genetics ; immunology ; pathology ; T-Lymphocytes ; metabolism ; pathology ; Transforming Growth Factor beta ; genetics ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo investigate if granulocyte-macrophage colony stimulating factor (GM-CSF) gene-modified dendritic cells (DC) enhance antitumor immunity in vitro.

METHODSMice were injected with chemokine ligand 3 (CCL3) via the tail vein. Fresh B220(-)CD11c(+) cells were sorted from the peripheral blood mononuclear cells (PBMCs) and cultured into DCs by cytokines.DCs were transfected with AdGM-CSF gene at different ratios of multiplicity of infection (MOI) to determine the optimal gene transfection conditions, and the expression of GM-CSF was detected after transfection. The variation of GM-CSF gene-modifiedDCs were analyzed by morphological examination, phenotype analysis, and mixed lymphocyte reaction (MLR).DCs were loaded with gastric cancer antigen obtained by freezing and thawing method. The killing effect of DCs vaccine-stimulated T lymphocytes on gastric cancer cells was assessed by MTT assay. INF-gamma production was determined with the INF-gamma ELISA kit.

RESULTSB220(-)CD11c(+) cells increased obviously after CCL3 injection. The ELISA results showed that after GM-CSF gene modification, DCs could produce high level of GM-CSF. When DCs were transfected with AdGM-CSF gene at MOI equal to 100, the GM-CSF level in culture supernatants reached saturation [(130.00 +/- 12.61) pg/ml]. After GM-CSF gene-modification, DCs tend to be more maturated as detected by morphological observation and phenotype analysis. At the same time, the capacity of activating the proliferation of allogeneic T lymphocytes was enhanced greatly. T lymphocytes stimulated by DCs transfected with GM-CSF gene showed a specific killing effect on gastric carcinoma cells and produced high level of INF-gamma [(1245.00 +/- 13.75) pg/ml].

CONCLUSIONAfter GM-CSF gene modification, DCs can produce high level of GM-CSF, which tend to be more maturated, and the capacity of activating the proliferation of allogeneic T lymphocytes is enhanced greatly. GM-CSF gene modified DCs can induce specific CTL to target tumor cells in vitro.

Adenoviridae ; genetics ; Animals ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Cell Proliferation ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; metabolism ; Histocompatibility Antigens Class II ; metabolism ; Interferon-gamma ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; Stomach Neoplasms ; immunology ; metabolism ; pathology ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Transfection