ABSTRACT
To explore the expression spectra of apoptosis-related gene pnas-2 in normal tissues and acute leukemia (AL) patient tissues, the expressions of pnas-2 gene in tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, leukocyte, bone marrow and fetal liver were detected by Northern blot. The expressions of pnas-2 in samples including 44 de novo, 9 non-CR, 27 CR and 12 relapsed AL patients were measured by real-time RT-PCR and Northern blot, and the expression levels of pnas-2 in normal and tumor tissues from 31 patients with malignancies were also detected. The results showed that pnas-2 was not expressed in the most tissues except in placenta. The results of real-time PCR indicated that pnas-2 expressions in samples of de novo, non-CR and relapsed patients ware significantly higher than that in CR, tumor tissues and normal tissues. In serial monitoring of 7 AL patients, the expression level of pnas-2 was high at first visit examination, but remarkably decreased after remission, and the pnas-2 expression level increased again when relapsed. It is concluded that the pnas-2 is specifically up-regulated in acute leukemia patients, which might be an oncogene and participate in leukemogenesis.
Subject(s)
Humans , Acute Disease , Apoptosis , Genetics , Apoptosis Regulatory Proteins , Genetics , Metabolism , Biomarkers, Tumor , Genetics , Gene Expression Regulation, Leukemic , Leukemia , PathologyABSTRACT
The study was purposed to explore the correlation between apoptosis-related gene pnas-2 and leukemia. The RT-PCR was performed to detect the expression levels of pnas-2 gene in NB4, K562, U937 cells before and after treatment with AS(4)S(4), and to analysis the expression change of pnas-2 gene in bone marrow cells from patients with acute leukemia before and after chemotherapy. The results showed that the expression of pnas-2 gene in arsenic sulfide treated NB4 cells was down regulated in time-dependent manner, but the same outcome in K562 and U937 cells after being treated with AS(4)S(4) was not found. The positive expression rate of pnas-2 in cells from untreated patients with acute leukemia was 100%, and was significantly higher than that in normal control group. After chemotherapy, the expression was negative in complete remission patients, whereas in no-remission patients there were no significant differences of expression of pnas-2 before and after treatment. It is concluded that the pnas-2 gene may be closely related with apotosis of arsenic sulfide treated APL cells, and may consider as a molecular biological remission marker in acute leukemia.
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Arsenicals , Pharmacology , K562 Cells , Leukemia , Pathology , Leukemia, Myeloid, Acute , Drug Therapy , Genetics , Pathology , Sulfides , Pharmacology , Tumor Cells, Cultured , U937 CellsABSTRACT
Objective Clinical significance of using ELISA to determine ?-amyloid(A?)_(1-42) antibody levels in the sera of patients with Alzheimer's disease(AD).Methods 96 wells PVC plate was coated with A?_(1-42)peptide.Serum of AD patient was competing with mouse A?_(1-42)monoclonal antibody in this assay.The second antibody was horseradish peroxidase(HRP)conjugated goat anti-mouse IgG.Serum A?_(1-42)antibody levels were determined by ELISA.Results The sensitivity of this assay was about 1 ng/ml.The recovery rate of this test was between 96.5% and 104.7%.The residual A?_(1-42)antibody levels in human serum or horse serum after A?_(1-42)antibody was removed by absorption were less than 1 ng/ml. Serum A?_(1-42)antibody levels in 37 AD patients[(5.1?1.9)ng/ml]were remarkably lower than those in normal people[(12.6?3.3)ng/ml,P