Apoptosis ; Down-Regulation ; Electron Transport Complex IV ; chemistry ; genetics ; metabolism ; Humans ; Mitochondria ; metabolism ; Mutation ; Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo construct hu-PBL/SCID chimeras and to investigate the development of lymphoma and oncogenicity of the Epstein-Barr virus (EBV).

METHODSHuman peripheral blood lymphocytes (PBLs) were isolated from healthy adult donors and transplanted intraperitoneally into severe combined immunodeficient (SCID) mice. Mice with hu-PBL engraftment from healthy EBV seronegative donors were injected intraperitoneally with EBV-containing supernatant from suspension culture of B95-8 cell line (active infection), whereas mice receiving lymphocytes from healthy EBV seropositive donors were not re-infected with B95-8 derived EBV (latent infection). Pathological examination and molecular analysis were performed on experimental animals and induced neoplasms.

RESULTSIn the early stage of this experiment, 12 mice died of acute graft-versus-host disease, mortality was 34.3% (12/35 mice) with an average life span of 17.5 days. In 19 survival hu-PBL/SCID chimeric recipients from 12 healthy donors, tumor incidence was 84.2% (16/19 mice). The average survival time of tumor-bearing mice was 65.5 days. EBV-related neoplasms in SCID mice were nodular tumors with aggressive and fatal features. Histological morphology of tumors exhibited diffuse large cell lymphomas. Immunohistochemistry revealed that LCA (CD45) and L26 (CD20) were positive, but both PS1 (CD3) and UCHL-1 (CD45RO) were negative, and EBV products ZEBRA, LMP1, and EBNA2 were expressed in a small number of tumor cells. EB virus particles were seen in the nuclei of some tumor cells by electron microscopy, and EBV DNA could be amplified in the tumor tissues by PCR. In situ hybridization indicated that the nuclei of tumor cells contained human-specific Alu sequence.

CONCLUSIONSEBV-induced tumors were human B-cell malignant lymphomas. We obtained direct causative evidence dealing with EBV-associated tumor deriving from normal human cells.

Adult ; Animals ; Antigens, CD20 ; metabolism ; Chimera ; Epstein-Barr Virus Infections ; immunology ; virology ; Graft vs Host Disease ; prevention & control ; virology ; Herpesvirus 4, Human ; physiology ; Humans ; Leukocyte Common Antigens ; metabolism ; Leukocyte Transfusion ; methods ; Lymphoma, B-Cell ; immunology ; virology ; Lysosomal-Associated Membrane Protein 1 ; metabolism ; Mice ; Mice, SCID

OBJECTIVETo obtain allelic loss mapping and define the minimal lost region on chromosome 1q21 in gastric carcinomas, and explore role of 1q21 loss of heterozygosity (LOH) in the development and progression of gastric carcinogenesis.

METHODSUsing 7 high-density microsatellite markers and PCR method, lq21 LOH was analyzed in 30 paired specimens of fresh gastric carcinoma, and the relation between 1q21 LOH and the clinicopathological features of the malignancy was tested.

RESULTSThe LOH frequency on chromosome 1q21 from these gastric carcinoma tissues reached 60% (18/30). The LOH frequencies of the microsatellite markers D1S514, D1S2696, D1S498, D1S305, D1S2624, D1S2635 and D1S2702 were 13.3%, 10%, 20%, 23.3%, 33.3%, 40% and 23.3%, respectively. The minimal lost region on 1q21 LOH in the gastric carcinoma tissues was located in the region between D1S2624 and D1S2707, in the vicinity of D1S2635. No significant relations of 1q21 LOH to the patients' age, gender, location of the primary foci, clinical staging, or the tumor differentiation were noted (P>0.05), but 1q21 LOH was correlated to lymph node metastases of the malignancy (P<0.05).

CONCLUSIONHigher frequency of 1q21 LOH occurs in gastric carcinoma cells, suggesting the presence of potential tumor suppressor genes closely associated with gastric carcinogenesis near the region of D1S2635.

Adenocarcinoma ; genetics ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; genetics ; Female ; Humans ; Loss of Heterozygosity ; Lymphatic Metastasis ; Male ; Microsatellite Repeats ; Middle Aged ; Stomach Neoplasms ; genetics