OBJECTIVE: To explore the relationship between the expression levels of JARID1B,Hes1 and MMP-9 genes and the stages of chronic myelogenous leukemia(CML) and the curative effect of imatinib mesylate (IM). METHODS: Peripheral blood samples of 15 cases of CML in chronic phase and 10 cases of CML in progressive phase were collected from the Hematology Department of Taihe Hospital affiliated to Hubei University of Medicine and 15 cases of healthy people in the Physical Examination Center. CML patients were divided into effective group and ineffective group based on the efficacy after treatment with IM, then real-time PCR was used to detect the expression levels of JARID1B, Hes1 and MMP-9 mRNA, finally, the differences in the level of gene expression and their correlations with CML stages and IM curative efficacy were analysed. RESULTS: The expression levels of Hes1 and MMP-9 in initially diagnosed patients in chronic and progressive phase without IM treatment were significantly higher than those of health people（P＜0.05）. There was no significant difference in the expression level of JARID1B between chronic phase patients and health people（P＞0.05）, but the expression level of JARID1B in the progressive phase patients was higher than that of health people （P＜0.05）. The expression levels of JARID1B and Hes1 in the IM-effective group were not significantly different from those in the IM-ineffective group (P=0.85,P=0.82）, while the expression level of MMP-9 in the IM-effective group [JP2]was significantly lower than that in the IM-ineffective group（P＜0.05）. CONCLUSION The expression levels of JARID1B Hes1 and MMP-9 relate with the different phase of CML; The expression levels of JARID1B and Hes1 have not significant relationship with IM curative efficacy, the MMP-9 gene expression level relates with IM curative efficacy.
Antineoplastic Agents ; therapeutic use ; Humans ; Imatinib Mesylate ; therapeutic use ; Jumonji Domain-Containing Histone Demethylases ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Matrix Metalloproteinase 9 ; Nuclear Proteins ; Repressor Proteins ; Transcription Factor HES-1

Animals ; Epstein-Barr Virus Nuclear Antigens ; immunology ; Immunization ; Mice ; Mice, Inbred BALB C ; Viral Matrix Proteins ; immunology ; Viral Vaccines

BACKGROUND: Tetrabromobisphenol A (TBBPA), one of the most widely used brominated flame-retardants, is a representative persistent organic pollutants group. Studies on TBBPA toxicity have been conducted using various target cells; however, few studies have investigated TBBPA toxicity in bone cells. Therefore, this study investigated the in vitro effects of TBBPA on osteoclasts, a cell type involved in bone metabolism. METHODS: RAW264.7 cells were cultured in medium containing 50 ng/mL receptor activator of nuclear factor kappa B ligand (RANKL) and varying concentrations of TBBPA. To evaluate the effects of TBBPA on the differentiation and function of osteoclasts, osteoclast-specific gene expression, tartrate-resistant acid phosphatase (TRAP) activity, bone resorbing activity, mitochondrial membrane potential (MMP) and mitochondrial superoxide were measured. RESULTS: The presence of 20 μM TBBPA significantly increased TRAP activity in RANKL-stimulated RAW264.7 cells, the bone resorbing activity of osteoclasts, and the gene expression of Akt2, nuclear factor of activated T-cells cytoplasmic 1, and chloride channel voltage-sensitive 7. However, TBBPA treatment caused no change in the expression of carbonic anhydrase II, cathepsin K, osteopetrosis-associated transmembrane protein 1, Src, extracellular signal-related kinase, GAB2, c-Fos, or matrix metalloproteinase 9. Furthermore, 20 μM TBBPA caused a significant decrease in MMP and a significant increase in mitochondrial superoxide production. CONCLUSION: This study suggests that TBBPA promotes osteoclast differentiation and activity. The mechanism of TBBPA-stimulated osteoclastogenesis might include increased expression of several genes involved in osteoclast differentiation and reactive oxygen species production.
Acid Phosphatase ; Carbonic Anhydrase II ; Cathepsin K ; Chloride Channels ; Cytoplasm ; Gene Expression ; In Vitro Techniques ; Matrix Metalloproteinase 9 ; Membrane Potential, Mitochondrial ; Metabolism ; Osteoclasts ; Phosphotransferases ; RANK Ligand ; Reactive Oxygen Species ; Receptor Activator of Nuclear Factor-kappa B ; Superoxides ; T-Lymphocytes

BACKGROUND: Kaempferol (3,4′,5,7-tetrahydroxyflavone) is a flavonoid known to have a wide range of pharmacological activities. The 3-OH group in flavonoids has been reported to determine antioxidant activities. OBJECTIVE: We tested whether kaempferol can affect the expression of integrins and the stem cell fate of interfollicular epidermal stem cells. METHODS: Skin equivalent (SE) models were constructed, and the expression levels of stem cell markers and basement membrane-related antigens were tested. The immunohistochemical staining patterns of integrins, p63, and proliferating cell nuclear antigen (PCNA) were compared between kaempferol- and apigenin-treated SE models. Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the mRNA expression of integrins. RESULTS: Kaempferol increased the thickness of the epidermis when added to prepare SEs. In addition, the basal cells of kaempferol- treated SEs appeared more columnar. In the immunohistological study, the expression of integrins α6 and β1 and the numbers of p63- and PCNA-positive cells were markedly higher in the kaempferol-treated model. However, apigenin showed no effects on the formation of three-dimensional skin models. RT-PCR analysis also confirmed that kaempferol increased the expression of integrin α6 and integrin β1. CONCLUSION: Our findings indicated that kaempferol can increase the proliferative potential of basal epidermal cells by modulating the basement membrane. In other words, kaempferol can affect the fate of interfollicular epidermal stem cells by increasing the expression of both integrins α6 and β1. These effects, in particular, might be ascribed to the 3-OH group of kaempferol.
Apigenin ; Basement Membrane ; Epidermis ; Extracellular Matrix ; Flavonoids ; Integrins ; Proliferating Cell Nuclear Antigen ; RNA, Messenger ; Skin ; Stem Cells*

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-β1 and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3DTN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.
Bone Regeneration ; Cell Proliferation ; Cell- and Tissue-Based Therapy ; Chondrogenesis ; Collagen Type II ; Extracellular Matrix ; Hydrogel ; Hydrogels ; Mesenchymal Stromal Cells ; Osteocalcin ; Osteogenesis ; Polymers ; Proliferating Cell Nuclear Antigen ; Tissue Engineering ; Water

BACKGROUND: It is difficult to distinguish between actinic cheilitis and lichen planus histologically, because both types of lesions exhibit variable degrees of epidermal dysplasia and dermal lichenoid inflammation. There is currently no consensus on suitable immunohistochemical markers for distinguishing these 2 conditions. OBJECTIVE: This study aims to determine histological features and immunohistochemical markers that could be used to differentiate actinic cheilitis from lichen planus. METHODS: Fifteen cases of actinic cheilitis and 11 cases of lichen planus of the lips were included in the study. Histological changes such as parakeratosis, hyperkeratosis, atrophy, acanthosis, ulceration, necrosis, dermal solar elastosis, degrees of epidermal dysplasia and dermal inflammatory cell infiltration were examined. Verhoeff-van Gieson stained sections were quantified for the degree of elastosis using computer software. The following immunohistochemical markers were stained for: bcl-2, Ki-67, proliferating cell nuclear antigen, indoleamine 2, 3-dioxygenase, matrix metalloproteinase-3, matrix metalloproteinase-9, CD4, CD8, c-kit, and prolyl-4-hydroxylase. RESULTS: The only histologically appreciable difference between the diseases was the degree of epidermal dysplasia. No differences were observed with respect to solar elastosis using the Verhoeff-van Gieson stain. We found that cell proliferation markers such as proliferating cell nuclear antigen and Ki-67 were more highly expressed in actinic cheilitis than in lichen planus. In addition, the number of c-kit-positive cells observed in actinic cheilitis was significantly higher than in lichen planus. The expression levels of the other tested markers were not significantly different between the 2 diseases. CONCLUSION: The immunohistochemical markers proliferating cell nuclear antigen, Ki-67, and c-kit may help to differentiate actinic cheilitis from lichen planus of the lips.
Actins* ; Atrophy ; Cell Proliferation ; Cheilitis* ; Consensus ; Inflammation ; Lichen Planus* ; Lip* ; Matrix Metalloproteinase 9 ; Necrosis ; Parakeratosis ; Proliferating Cell Nuclear Antigen ; Ulcer

OBJECTIVETo put the insight into the trichloroethylene (TCE)-induced effect on the differential expression of subcellular proteins in human normal liver cell line (L-02).

METHODSThe membrane proteins and nuclear proteins of TCE-treated (8.0 mmol/L) group and controls were extracted by subcellular proteome extraction kit, respectively. The TCE-induced differentially expressions were analyzed by a two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization tandem time-of-flight spectrometry (MALDI-TOF-MS). Bioinformatics analysis was used to reveal the biological processes and predict transmembrane domains of differential expressed proteins. The expression of ATP synthase subunit beta (ATP5B), heterogeneous nuclear ribonucleoprotein H2 (hnRNP H2) and far up steam element-binding protein 1 (FUBP1) were measured under TCE treatment by Western blot.

RESULTSAfter TCE treatment for 24 h in L-02 cells, 14 membrane proteins and 18 nuclear proteins were identified as differential expression. After treated with TCE in concentrations of 0, 2.0, 4.0 and 8.0 mmol/L for 24 h, the relative levels of ATP5B expression were 1.00±0.03, 1.21±0.14, 1.25±0.12 and 1.48±0.17 (F = 8.51, P = 0.007), the relative levels of hnRNP H2 expression were 1.00±0.09, 1.22±0.15, 1.43±0.21, 1.53±0.17 (F = 6.57, P = 0.015), respectively; the relative levels of FUBP1 expression were 1.00±0.11, 0.91±0.07, 0.73±0.04 and 0.67±0.03 (F = 15.81, P = 0.001), respectively, which were consistent with the results in proteomics. The bioinformatics analysis showed that the most dominant biological process were involved in RNA processing (10 proteins, P = 2.46×10(-6)), especially in RNA splicing (9 proteins, P = 1.77×10(-7)).

CONCLUSIONThe exposure of TCE could alter the expression of membrane proteins and nuclear proteins in L-02 cells. These abnormal expressed proteins involved in RNA splicing would provide novel clues for further understanding of TCE-induced hepatotoxicity.

Blotting, Western ; Cell Line ; DNA Helicases ; DNA-Binding Proteins ; Hepatocytes ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H ; Humans ; Mitochondrial Proton-Translocating ATPases ; Proteome ; Proteomics ; RNA Processing, Post-Transcriptional ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trichloroethylene

Latent Epstein-Barr virus (EBV) infection has a substantial role in causing many human disorders. The persistence of these viral genomes in all malignant cells, yet with the expression of limited latent genes, is consistent with the notion that EBV latent genes are important for malignant cell growth. While the EBV-encoded nuclear antigen-1 (EBNA-1) and latent membrane protein-2A (LMP-2A) are critical, the EBNA-leader proteins, EBNA-2, EBNA-3A, EBNA-3C and LMP-1, are individually essential for in vitro transformation of primary B cells to lymphoblastoid cell lines. EBV-encoded RNAs and EBNA-3Bs are dispensable. In this review, the roles of EBV latent genes are summarized.
Epstein-Barr Virus Infections/complications/virology ; Epstein-Barr Virus Nuclear Antigens/genetics/metabolism ; *Genes, Viral ; Herpesvirus 4, Human/*physiology ; Humans ; MicroRNAs/genetics ; Neoplasms/etiology ; Protein Binding ; RNA, Viral/genetics ; Viral Matrix Proteins/genetics/metabolism ; *Virus Latency

OBJECTIVETo investigate the effect of RbAp48 knockdown on the migration and invasion of human cervical cancer cells and explore the mechanism.

METHODSA small interference RNA (siRNA) was used to knock down the expression of RbAp48 in MS751 cells. The changes in cell migration and invasion were evaluated using wound healing assay and Transwell assay, respectively, and the expressions of RbAp48, vimentin, N-cadherin, E-cadherin, Snail, Twist, MMP-2 and TIMP-2 were determined with Western blotting.

RESULTSAfter siRNA-mediated RbAp48 knockdown, MS751 cells showed a significantly reduced expression of RbAp48 with significantly suppressed cell migration and invasion (P<0.01). RbAp48 knockdown induced obvious down-regulation of the expressions of interstitial cell phenotype proteins vimentin, N-cadherin, and MMP-2 and up-regulation of epithelial cell phenotype proteins E-cadherin and TIMP-2, suggesting the inhibition of epithelial- mesenchymal transition of the cells. The expressions of Snail and Twist were significantly down-regulated in the cells following RbAp48 knockdown.

CONCLUSIONKnockdown of RbAp48 can significantly inhibit epithelial-mesenchymal transition and suppress the migration and invasion of cervical cancer cell line MS751, the mechanism of which may involve the down-regulation of Snail and Twist expressions.

Antigens, CD ; metabolism ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Movement ; Down-Regulation ; Epithelial-Mesenchymal Transition ; Female ; Gene Knockdown Techniques ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Neoplasm Invasiveness ; Nuclear Proteins ; metabolism ; RNA, Small Interfering ; Retinoblastoma-Binding Protein 4 ; genetics ; Snail Family Transcription Factors ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Transcription Factors ; metabolism ; Twist-Related Protein 1 ; metabolism ; Up-Regulation ; Uterine Cervical Neoplasms ; pathology ; Vimentin ; metabolism

OBJECTIVETo establish a model of chondrocyte degeneration in vitro.

METHODSChondrocytes were isolated from articular cartilages of newly born SD rats by digestion with typeⅡ collagenase. The chondrocytes were cultured with H-DMEM medium containing 10%FBS, 50 ng/mL IL-1β+10%FBS, 2.5% rat serum and 5% rat serum, respectively; and the chondrocytes at passage one were used in the experiments. The morphology changes were investigated under phase contrast microscope after chondrocytes were treated with rat serum and IL-1β. Proliferation of chondrocytes was detected by MTT method. The protein expression levels of PCNA, typeⅡ collagen and MMP-13 were examined by Western blotting. The levels of ADAMTS5, MMP-9, Aggrecan and SOX-9 mRNA were detected by real-time PCR.

RESULTSThe cell morphology was changed from polygon to spindle in both rat serum groups and IL-1β group, and the proliferation of chondrocytes in these groups was much higher than that in control group. The results showed that the expression levels of typeⅡ collagen, Aggrecan and SOX-9 decreased while the expression levels of MMP-13, MMP-9 and ADMATS5 were up-regulated in rat serum and IL-1β-treated groups compared with control group.

CONCLUSIONThe results indicate that rat serum can induce chondrocyte degeneration and may be used for osteoarthritis model in vitro.

ADAM Proteins ; metabolism ; ADAMTS5 Protein ; Aggrecans ; metabolism ; Animals ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; pathology ; Collagen Type II ; metabolism ; Disease Models, Animal ; Interleukin-1beta ; pharmacology ; Matrix Metalloproteinase 13 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Osteoarthritis ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; metabolism ; Serum ; Up-Regulation