microRNAs are a class of short non-coding RNAs containing about 19-22 nucleotides that regulate target gene expression through post-transcriptional repression or mRNA degradation, and involved in a variety of biological processes, such as cellular differentiation, proliferation, apoptosis and metabolism. microRNA-182 (miR-182), belonging to miR-183/96/182 cluster that consists of miR-182, -183, and -96, highly expresses in many cells and tissues, including osteoblasts, lymphocytes, adipocytes, retina, inner ear, etc. The recent studies of miR-182 highlighted its multiple important roles in differentiation, development, and functional maintenance in the cells and tissues. The dysregulation of miR-182 is associated with occurrence and development of many diseases, such as retinopathy, autoimmune diseases, cancers, obesity and diabetes. This review summarizes recent research progresses on the roles and mechanisms of miR-182 in cellular function and diseases.
Apoptosis ; Cell Differentiation ; MicroRNAs ; genetics ; Neoplasms ; Osteoblasts

OBJECTIVE: To investigate the expression of miR-451 during erythroid differentiation and its correlation with hematological diseases. METHODS: The expression of miR-451 in erythroid differentiation of mouse hematopoietic stem cells (derived from fetal liver) was analyzed by cell culture, flow cytometry, magnetic bead sorting and qRT-PCR. The expression of miR-451 during erythroid differentiation of mouse erythroid leukemia cells (MEL) was analyzed by cell culture and qRT-PCR. The expression of miR-451 in peripheral blood of mice was detected by qRT-PCR, and the expression of miR-451 in fetal liver (14.5 days) was analyzed by microarray. The nucleated erythroid cells from bone marrow of wild type (WT) mice and β-thalassemia (β-thal) mice were sorted by flow cytometry, and the levels of miR-451 and erythroid genes were detected by qRT-PCR. The expression of miR-451 in peripheral blood of patients with clinical hematological diseases was detected by qRT-PCR. RESULTS: During the differentiation of mouse hematopoietic stem cells (derived from fetal liver) and MEL cells, the expression levels of miR-451 increased gradually. Compared with WT mice, the expression levels of miR-451 in peripheral blood, 14.5-day fetal liver cells and nucleated erythroid cells (sorted from bone marrow) of β-thal mice were significantly increased(P<0.05). Many erythroid differentiation genes in nucleated erythroid cells (sorted from bone marrow) of β-thal mice decreased. Compared with healthy controls, the expression levels of miR-451 was increased in peripheral blood of patients with β-thalassemia and iron deficiency anemia, while the expression levels of miR-451 was decreased in patients with aplastic anemia and myelodysplastic syndrome. CONCLUSION During erythroid differentiation, the expression levels of miR-451 increases gradually. In hematological diseases, the expression levels of miR-451 is different from that of normal controls, which is expected to become an auxiliary diagnostic index for clinical hematological diseases.
Mice ; Animals ; beta-Thalassemia/genetics* ; Cell Differentiation ; MicroRNAs/genetics*

Animals ; Cell Differentiation ; genetics ; physiology ; Humans ; MicroRNAs ; genetics ; Ossification, Heterotopic ; genetics ; Signal Transduction ; genetics ; physiology

C-fos is a transcription factor that plays an important role in cell proliferation, differentiation and tumor formation. The aim of this study was to clone the goat c-fos gene, clarify its biological characteristics, and further reveal its regulatory role in the differentiation of goat subcutaneous adipocytes. We cloned the c-fos gene from subcutaneous adipose tissue of Jianzhou big-eared goats by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed its biological characteristics. Using real-time quantitative PCR (qPCR), we detected the expression of c-fos gene in the heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi and subcutaneous adipocytes of goat upon induced differentiation for 0 h to 120 h. The goat overexpression vector pEGFP-c-fos was constructed and transfected into the subcutaneous preadipocytes for induced differentiation. The morphological changes of lipid droplet accumulation were observed by oil red O staining and bodipy staining. Furthermore, qPCR was used to test the relative mRNA level of the c-fos overexpression on adipogenic differentiation marker genes. The results showed that the cloned goat c-fos gene was 1 477 bp in length, in which the coding sequence was 1 143 bp, encoding a protein of 380 amino acids. Protein structure analysis showed that goat FOS protein has a basic leucine zipper structure, and subcellular localization prediction suggested that it was mainly distributed in the nucleus. The relative expression level of c-fos was higher in the subcutaneous adipose tissue of goats (P < 0.05), and the expression level of c-fos was significantly increased upon induced differentiation of subcutaneous preadipocyte for 48 h (P < 0.01). Overexpression of c-fos significantly inhibited the lipid droplets formation in goat subcutaneous adipocytes, significantly decreased the relative expression levels of the AP2 and C/EBPβ lipogenic marker genes (P < 0.01). Moreover, AP2 and C/EBPβ promoter are predicted to have multiple binding sites. In conclusion, the results indicated that c-fos gene was a negative regulatory factor of subcutaneous adipocyte differentiation in goats, and it might regulate the expression of AP2 and C/EBPβ gene expression.
Animals ; Goats/genetics* ; Cell Differentiation/genetics* ; Adipogenesis/genetics* ; Gene Expression Regulation ; Proteins/genetics* ; Cloning, Molecular

The aim of this study was to clone the goat RPL29 gene and analyze its effect on lipogenesis in intramuscular adipocytes. Using Jianzhou big-eared goats as the object, the goat RPL29 gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR), the gene structure and expressed protein sequence were analyzed by bioinformatics, and the mRNA expression levels of RPL29 in various tissues and different differentiation stages of intramuscular adipocytes of goats were detected by quantitative real-time PCR (qRT-PCR). The RPL29 overexpression vector pEGFP-N1-RPL29 constructed by gene recombination was used to transfect into goat intramuscular preadipocytes and induce differentiation. Subsequently, the effect of overexpression of RPL29 on fat droplet accumulation was revealed morphologically by oil red O and Bodipy staining, and changes in the expression levels of genes related to lipid metabolism were detected by qRT-PCR. The results showed that the length of the goat RPL29 was 507 bp, including a coding sequence (CDS) region of 471 bp which encodes 156 amino acid residues. It is a positively charged and stable hydrophilic protein mainly distributed in the nucleus of cells. Tissue expression profiling showed that the expression level of this gene was much higher in subcutaneous adipose tissue and inter-abdominal adipose tissue of goats than in other tissues (P < 0.05). The temporal expression profile showed that the gene was expressed at the highest level at 84 h of differentiation in goat intramuscular adipocytes, which was highly significantly higher than that in the undifferentiated period (P < 0.01). Overexpression of RPL29 promoted lipid accumulation in intramuscular adipocytes, and the optical density values of oil red O staining were significantly increased (P < 0.05). In addition, overexpression of RPL29 was followed by a highly significant increase in ATGL and ACC gene expression (P < 0.01) and a significant increase in FASN gene expression (P < 0.05). In conclusion, the goat RPL29 may promote intra-muscular adipocyte deposition in goats by up-regulating FASN, ACC and ATGL.
Animals ; Lipogenesis/genetics* ; Adipogenesis/genetics* ; Goats/genetics* ; Adipocytes ; Cell Differentiation/genetics* ; Sequence Analysis ; Cloning, Molecular

Cell reprogramming is a progress in which the memory of a mature cell is erased and then the cell develops novel phenotype and function; ultimately, the fate of the cell changes. Cell reprogramming usually occurs at genes expression levels that no genomic DNA sequence change will be involved. By changing the programs of the genetic expressions of cells in terms of space and time, cell reprogramming alters the differentiation of cells and thus produces the required cells. Further research on cells reprogramming will elucidate the mechanisms that govern the cell development, and thus provides more information of the sources of seed cells used for regeneration medicine. More cells differentiated from many terminally differentiated cells will be obtained, which is extremely important for the understanding of molecular differentiation and for the development of cell replacement therapy. This article summarizes the classification, influencing factors, approaches and latest advances of cells reprogramming.
Animals ; Cell Dedifferentiation ; genetics ; Cell Differentiation ; genetics ; Cellular Reprogramming ; Gene Expression ; Humans ; Nuclear Transfer Techniques

OBJECTIVE: To obtain skin-derived induced pluripotent stem cells (iPSCs) from an Osteogenesis imperfecta (OI) patient carrying WNT1^c.677C>T mutation in order to provide a new cell model for investigating the underlying molecular mechanism and stem cell therapy for OI. METHODS: The pathogenic variant of the patient was identified by Sanger sequencing. With informed consent from the patient, skin tissue was biopsied, and primary skin fibroblasts were cultured. Skin fibroblasts were induced into iPSCs using Sendai virus-mediated non-genomic integration reprogramming method. The iPSC cell lines were characterized for pluripotency, differentiation capacity, and karyotyping assay. RESULTS: The patient was found to carry homozygous missense c.677C>T (p.Ser226Leu) mutation of the WNT1 gene. The established iPSC lines possessed self-renewal and capacity for in vitro differentiation. It also has a diploid karyotype (46,XX). CONCLUSION A patient-specific WNT1 gene mutation (WNT1^c.677C>T) iPSC line was established, which can provide a cell model for the study of OI caused by the mutation.
Humans ; Induced Pluripotent Stem Cells/pathology* ; Osteogenesis Imperfecta/genetics* ; Mutation ; Cell Differentiation/genetics* ; Cell Line

microRNAs (miRNAs) are small molecular non-coding RNA with 21-25 nucleotides in a variety of eukaryotic systems, and regulate gene expression at the post-transcriptional level by degrading or translational repressing target messenger RNA (mRNA). Many studies have showed the roles of miRNAs in normal lymphocytopoiesis, giving an interpretative key to the aberrant expression observed in human lymphoid malignancies. The recent advances of understanding the roles of miRNAs in lymphoid malignancies show that miRNAs as tumoral biomarkers can effectively be used for diagnosis, prognosis, and prediction of response to therapy. This review focuses the roles of miRNA in development and differentiation of lymphocytes and the relation of miRNA to lymphoid malignancies.
Cell Differentiation ; Humans ; Lymphocytes ; cytology ; Lymphopoiesis ; genetics ; MicroRNAs ; Neoplasms ; genetics ; pathology ; RNA, Messenger ; genetics

This study aimed to explore the effect of miR-23b-3p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-23b-3p plays its roles via targeting the PDE4B gene. Based on the pre-transcriptome sequencing data obtained previously, the miR-23b-3p, which was differentially expressed in goat intramuscular adipocytes before and after differentiation, was used as an entry point. real-time quantitative-polymerase chain reaction (qPCR) was used to detect the expression pattern of miR-23b-3p during the differentiation of goat intramuscular preadipocytes. The effects of miR-23b-3p on adipose differentiation and adipose differentiation marker genes were determined at the morphological and molecular levels. The downstream target genes of miR-23b-3p were determined using bioinformatics prediction as well as dual luciferase reporter assay to clarify the targeting relationship between miR-23b-3p and the predicted target genes. The results indicated that overexpression of miR-23b-3p reduced lipid droplet accumulation in goat intramuscular adipocytes, significantly down-regulated the expression levels of adipogenic marker genes AP2, C/EBPα, FASN, and LPL (P < 0.01). In addition, the expressions of C/EBPβ, DGAT2, GLUT4 and PPARγ were significantly downregulated (P < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of ACC, ATGL, AP2, DGAT2, GLUT4, FASN and SREBP1 were extremely significantly up-regulated (P < 0.01), and the expression levels of C/EBPβ, LPL and PPARγ were significantly up-regulated (P < 0.05). It was predicted that PDE4B might be a target gene of miR-23b-3p. The mRNA expression level of PDE4B was significantly decreased after overexpression of miR-23b-3p (P < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of PDE4B (P < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with PDE4B gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.
Animals ; MicroRNAs/metabolism* ; Goats/genetics* ; PPAR gamma/metabolism* ; Adipogenesis/genetics* ; Cell Differentiation/genetics* ; Luciferases ; RNA, Messenger

OBJECTIVETo study the potential mechanism of the new steroidal drug NSC67657 induced leukemic cells differentiation.

METHODSCell proliferation was assayed by MTT assay. Surface antigen CD14 on THP-1 cells treated by NSC67657 at different time different concentration, was detected by flow cytometry (FCM). The expression of beta-catenin- interacting protein 1 (ICAT) gene and protein were detected by RT-PCR and Western blot. Eukaryotic expressing vector pDsRed-ICAT was constructed and transfected into HL60 cell line. FCM, Wright's staining and electronmicroscope were employed to analyse the differentiation of transfected THP-1 cells after they were treated with NSC67657 for 24 hours.

RESULTSThe proliferation of THP-1 cells was significantly inhibited by NSC67657 treatment. The level of CD14 expression was elevated in line with the increasing drug concentration and treatment time. 10 µmol/L NSC67657 treatment for five days was the optimal condition for the induction of THP-1 cells differentiation, when the CD14(+) THP-1 cells were more than 90%. Morphological study indentified the THP-1 cells of monocytic differentiation. The eukaryotic expressing vector pDSRed-ICAT was successfully constructed, and almost 90% positive clone could be obtained after G418 screening. Electro-transfection was employed for transfecting the vector into THP-1 cells. After the transfection the expression of ICAT gene and protein was increased. On the NSC67657 treatment, there was not significant difference in CD14 expression on transfected THP-1 cells compared to that on the control groups. After 24 h treatment, the transfected THP-1 cells remained in early differentiated stage.

CONCLUSIONNSC67657 can induce THP-1 cell to monocytic differentiation and activate the expression of ICAT gene, but overexpression of ICAT itself is not sufficient to induce such differentiation.

Cell Differentiation ; genetics ; Cell Line, Tumor ; Cell Proliferation ; HL-60 Cells ; Humans ; RNA, Messenger ; genetics ; Transfection ; beta Catenin ; genetics