ABSTRACT
<p><b>OBJECTIVE</b>To elucidate the role of let-7d in regulating the biological behavior of ovarian cancer cells and their expressions of HMGA2 and ras proteins.</p><p><b>METHODS</b>The pre-let-7d sequence was synthesized and inserted into pcDNA6.2GW/EmGFPmiR and transfected into ovarian cancer IGROV1 cells to cause pre-let-7d overexpression. Real-time quantitative RT-PCR was employed to examine the expression levels of let-7d miRNA and HMGA2 mRNA, and Western blotting was performed to detect the expressions of HMGA2 and ras protein in the transfected cells. The effect of pcDNA6.2GW-let-7d transfection on IGROV1 cell proliferation was determined using MTT assay and the cell apoptosis rate was measured using flow cytometry.</p><p><b>RESULTS</b>The eukaryotic expression vector containing the target gene let-7d was successfully constructed and transfected into IGROV1 cells. The transfected cells showed a marked reduction of HMGA2 expression but a less obvious down-regulation of ras expression. Transfection with pcDNA6.2GW-let-7d to suppress the expression of HMGA2 caused alterations of the phenotype of IGROV1 cells shown by a reduced proliferative activity and increased cell apoptosis.</p><p><b>CONCLUSION</b>Let-7d plays an important role in altering the malignant cell phenotype of ovarian cancer IGROV1 cells by regulating the expression of HMGA2.</p>
Subject(s)
Female , Humans , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , HMGA2 Protein , Genetics , Metabolism , MicroRNAs , Genetics , Ovarian Neoplasms , Genetics , Metabolism , Pathology , Plasmids , ras Proteins , Genetics , MetabolismABSTRACT
Objective To investigate the molecular characterization of a Chinese pedigree with rare β thalassemia genotype.Methods Phenotypic analysis was performed using standard hematological tests to measure red blood cell parameters, including RBC,Hb,MCV,MCH,MCHC and RDW.SPIFE automatic Hb agarose gel electrophoresis instrument was used to measure hemoglobin fraction Hb A,Hb A2 and Hb F.The alleles of β thalassemia mutation were determined by RDB assay, and then cloning and sequencing were performed to define the mutation sites.Results The proband and his father had typical microcytic hypochromic anemia with low MCV and MCH(79.8, 63.1 fl and 19.9, 20.9 pg, respectively) and high level of Hb A2 (5.66% and 5.60%, respectively).The proband′s mother had normal MCV and MCH. β thalassemia mutation analysis with RDB assay showed that the proband had thalassemia minor resulting from double mutations on one globin gene.One showed codons 41/42 (-TTCT) mutation and the other was CAP mutation from positions +40 to +43 in the promoter region.These two mutations were inherited from his father.The genotype of the proband and his father was β41/42、CAP/βA ,and the genotype of his mother was βA/βA.Conclusions It′s rare that double mutations occur on single β globin gene, with one mutation on CD41/42(-TTCT) and the other mutation from positions +40 to +43 relative to the mRNA cap site in the promoter region.The findings enrich knowledge of the mutation spectrum of β thalassemia.
ABSTRACT
Objective To investigate whether antenatal administration of dexamethasone would provide protection against cerebral hypoxic ischemic damage in asphyxiated fetal rats. Methods Fifty seven fetal rats of twenty day gestational age were randomly divided into five groups: sham operation (normal control, n =11), asphyxiated control (group E, n =10), and group D 1?D 2?D 3 of different timings of intravenous dexamethasone treatment (1 mg/kg) in pregnant SD rats ( n =15, 12, and 9 respectively). The times assigned for dexamethasone injection were thirty minutes before clamping, the time aroung clamping and the beginning of reperfusion respectively. Intra and extra cellular concentrations of calcium, sodium and potassium in fetal rat brains were measured in each group after thirty minutes of reperfusion. Results (1)Intracellular free calcium concentrations of fetal rat brains in group E,D 1, D 2 and D 3 were 552?94, 438?105, 445?57, and 456?110 nmol/L respectively, and all significantly higher than that in the normal control 315?87 nmol/L ( P