This study was aimed to quantitatively detect the level of maf-b mRNA in leukemia patients and evaluate its clinical significance. Real-time fluorescence quantitative PCR was used to detect the relative expression level of maf-b mRNA. The expression change of maf-b mRNA in various types of leukemia was analyzed. Then, the relationship of maf-b mRNA expression with laboratory index and the response to chemotherapy was analyzed. The results showed that maf-b mRNA expression level in acute myeloid leukemia (AML) patients was lower than that in normal group (p<0.01) and positively correlated with white blood cell count (p<0.01) and the expression of CD34 (p<0.01). There was no correlation between maf-b mRNA expression level and chemotherapy response in AML patients except for acute promyelocytic leukemia (APL). Maf-b mRNA expression levels in acute lymphoid leukemia (ALL) and chronic myeloid leukemia (CML) patients were also lower than that in normal group (p<0.01). It is concluded that there is low expression of maf-b gene in AML patients. Abnormal expression of maf-b correlates with abnormal proliferation of AML cells, which may be a new prognostic factor for AML.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cell Proliferation ; Child ; Female ; Humans ; Leukemia ; genetics ; pathology ; Leukemia, Myeloid, Acute ; genetics ; pathology ; MafB Transcription Factor ; genetics ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Young Adult

BACKGROUNDIn previous work, we suggested that some 2-aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells. Here, we reported the actions of another new 2-aminosteroid designated as H89712 on human leukemia cells.

METHODSCell colony counting and MTT assay were used to determine proliferation. Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation. RT-PCR was used to detect gene expression. Standard statistical method was used to analyze data.

RESULTSH89712 inhibited proliferation of HL-60 leukemia cells and the inhibition percentage in MTT assay was 18% at the dose of 10(-8) mol/L and 65% at the dose of 10(-5) mol/L, respectively. The inhibition for HL-60 in colony assay was 23% at the dose of 10(-8) mol/L and 96% at the dose of 10(-5) mol/L, respectively. H89712 also induced HL-60 cells toward macrophage-like differentiation. It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated HL-60 cells was about 9 times higher than that of the control at the dose of 10(-8) mol/L and 20 times higher than that of the control at the dose of 10(-5) mol/L respectively, and this action involved upregulation of MafB gene in HL-60 leukemia cells. On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 10(-8) mol/L and 88% at the dose of 10(-5) mol/L respectively. The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 10(-8) mol/L and 100% at the dose of 10(-5) mol/L respectively. H89712 also induced K562 cells toward erythroid-like differentiation and it was verified by flow cytometry that the percentage of positive CD71 expression in differentiated K562 cells was about 9 times higher than that of the control at the dose of 10(-8) mol/L and 16 times higher than that of the control at the dose of 10(-5) mol/L respectively. This action was related to upregulation of Egr-1 gene in K562 leukemia cells.

CONCLUSIONSOur results showed the important roles played by MafB in macrophage differentiation and Egr-1 in erythroid differentiation of human myeloid leukemia cells.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; DNA-Binding Proteins ; genetics ; Early Growth Response Protein 1 ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Immediate-Early Proteins ; genetics ; K562 Cells ; MafB Transcription Factor ; Oncogene Proteins ; genetics ; Steroids ; pharmacology ; Transcription Factors ; genetics ; Up-Regulation

Genetic alterations have been recognized as an important event in the carcinogenesis of gastric cancer (GC). We conducted high resolution bacterial artificial chromosome array-comparative genomic hybridization, to elucidate in more detail the genomic alterations, and to establish a pattern of DNA copy number changes with distinct clinical variables in GC. Our results showed some correlations between novel amplified or deleted regions and clinical status. Copy-number gains were frequently detected at 1p, 5p, 7q, 8q, 11p, 16p, 20p and 20q, and losses at 1p, 2q, 4q, 5q, 7q, 9p, 14q, and 18q. Losses at 4q23, 9p23, 14q31.1, or 18q21.1 as well as a gain at 20q12 were correlated with tumor-node-metastasis tumor stage. Losses at 9p23 or 14q31.1 were associated with lymph node status. Metastasis was determined to be related to losses at 4q23 or 4q28.2, as well as losses at 4q15.2, 4q21.21, 4q 28.2, or 14q31.1, with differentiation. One of the notable aspects of this study was that the losses at 4q or 14q could be employed in the evaluation of the metastatic status of GC. Our results should provide a potential resource for the molecular cytogenetic events in GC, and should also provide clues in the hunt for genes associated with GC.
Stomach Neoplasms/genetics/*pathology ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Receptors, Thyrotropin/genetics ; Nucleic Acid Hybridization/*methods ; Neoplasm Staging ; Middle Aged ; Male ; MafB Transcription Factor/genetics ; Lymphatic Metastasis/genetics ; Humans ; Genome, Human/genetics ; Gene Expression Regulation, Neoplastic ; Female ; Chromosomes, Human, Pair 20/genetics ; Chromosomes, Human, Pair 14/genetics ; *Chromosome Aberrations ; Aged, 80 and over ; Aged ; Adult

Aminoglycoside antibiotics, due to their strong antibacterial effects and broad antimicrobial spectra, have been very commonly used in clinical practice in the past half century. However, aminoglycoside antibiotics manifest severe ototoxicity and nephrotoxicity, and are one of top factors in hearing loss. In this study, three members of the aminoglycoside antibiotics family, gentamycin, neomycin and streptomycin, were chosen as the representatives to be investigated for their toxicity to the embryonic development and the larva hair cells in zebrafish, and also to their target genes associated with hearing-related genes. The results showed that: (1) the lethal effect of all three drugs demonstrated a significant dependence on concentration, and the severity order of the lethal effect was streptomycin > neomycin > gentamycin; (2) all the three drugs caused the larva trunk bending in resting state at 5 dpf (day past fertilization), probably due to their ototoxicity in the physical imbalance and postural abnormalities; (3) impairment and reducing of the hair cells were observed in all three cases of drug treatment; (4) four genes, eya1, val, otx2 and dlx6a, which play an important role in the development of hearing organs, showed differential and significant decrease of gene expression in a drug concentration-dependent manner. This study for the first time reports the relevance between the expression of hearing genes and the three ototoxic antibiotics and also proved the feasibility of establishing a simple, accurate, intuitive and fast model with zebrafish for the detection of drug ototoxicity.
Aminoglycosides ; toxicity ; Animals ; Anti-Bacterial Agents ; toxicity ; Embryonic Development ; drug effects ; Gene Expression Regulation ; Gentamicins ; toxicity ; Hair Cells, Auditory ; cytology ; drug effects ; Hearing Disorders ; chemically induced ; genetics ; metabolism ; Homeodomain Proteins ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Larva ; drug effects ; Lateral Line System ; drug effects ; MafB Transcription Factor ; metabolism ; Models, Animal ; Neomycin ; toxicity ; Nerve Tissue Proteins ; metabolism ; Nuclear Proteins ; metabolism ; Otx Transcription Factors ; metabolism ; Protein Synthesis Inhibitors ; toxicity ; Protein Tyrosine Phosphatases ; metabolism ; Streptomycin ; toxicity ; Zebrafish ; embryology ; Zebrafish Proteins ; metabolism