Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 17 de 17
Filter
1.
Article in Chinese | WPRIM | ID: wpr-878425

ABSTRACT

OBJECTIVES@#To investigate the expression of cyclophilin A (CyPA) in oral squamous cell carcinoma (OSCC) and explore the effect of downregulating the expression of CyPA gene on the proliferation and invasion of SCC-25 cells.@*METHODS@#A total of 77 cases of patients with OSCC were selected. The expression levels of CyPA proteins in OSCC and adjacent normal tissues were evaluated. SCC-25 cells were cultured and divided into the CyPA interference sequence group, negative control group, and blank group. The expression levels of CyPA mRNA and protein in cells were detected by using real-time fluorescent quantitative polymerase chain reaction and Western blot, respectively. Cell proliferation was detected by using methyl thiazolyl tetrazolium and plate colony formation assays. Cell invasion was detected by using Transwell assay.@*RESULTS@#The positive expression rate of CyPA protein in OSCC tissues was 76.62%, which was higher than that in adjacent tissues (@*CONCLUSIONS@#The CyPA protein is highly expressed in OSCC tissues, and the downregulation of CyPA gene expression in SCC-25 cells can reduce cell proliferation and inhibit cell invasion.


Subject(s)
Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cyclophilin A/genetics , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms , Humans , Mouth Neoplasms/genetics , Squamous Cell Carcinoma of Head and Neck
2.
Chinese Journal of Biotechnology ; (12): 605-611, 2020.
Article in Chinese | WPRIM | ID: wpr-827008

ABSTRACT

Cyclophilin A (CypA) is a widely distributed and highly conserved protein in organisms. It has peptidyl-prolyl cis/trans isomerase activity and is a receptor for cyclosporin A (CsA). Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. Seven types of coronaviruses are currently known to infect humans, among which SARS-CoV, MERS-CoV, and SARS-CoV-2 are fatal for humans. It is well established that CypA is essential for the replication of various coronaviruses such as SARS-CoV, CoV-229E, CoV-NL63, and FCoV. Additionally, CsA and its derivatives (ALV, NIM811, etc.) have obvious inhibitory effects on a variety of coronaviruses. These results suggest that CypA is a potential antiviral target and the existing drug CsA might be used as an anti-coronavirus drug. At the end of 2019, SARS-CoV-2 raged in China, which seriously theatern human health and causes huge economic lases. In view of this, we describe the effects of CypA on the replication of coronaviruses and the antiviral activities of its inhibitors, which will provide the scientific basis and ideas for the development of antiviral drugs for SARS-CoV-2.


Subject(s)
Antiviral Agents , Pharmacology , Therapeutic Uses , Betacoronavirus , Coronavirus , Coronavirus Infections , Drug Therapy , Epidemiology , Virology , Cyclophilin A , Cyclosporine , Chemistry , Pharmacology , Therapeutic Uses , Humans , Pandemics , Pneumonia, Viral , Drug Therapy , Epidemiology , Virology , SARS Virus , Virus Replication
3.
Article in Chinese | WPRIM | ID: wpr-815010

ABSTRACT

OBJECTIVE@#To explore the expression of CyPA and CD147 in rabbit models of vulnerable carotid atherosclerotic plaque and the therapeutic effect of atorvastatin.
@*METHODS@#Twenty-four male New Zealand rabbits were randomly divided into 3 groups. Eight rabbits were served as a normal diet group (Group A), and the remaining 16 rabbits underwent balloon-induced endothelial injury in the right carotid artery and thereafter were fed on high-cholesterol diet (1% cholesterol) for 12 weeks, then they were divided into 2 groups: a AS group (Group B), an atorvastatin group [Group C, 2.5 mg/(kg.d)]. 4 weeks later, plaque disrupture was triggered by China Russell's viper venom and histamine. Serum levels of TC, TG, LDL-C and HDL-C were measured at different timepoint. The damaged carotid arteries were collected to undergo pathological examination. The macrophage, expression of CyPA and CD147 were detected by immuno-histochemical analysis, and the mRNA levels of CyPA and CD147 were examined by reverse transcription polymerase chain reaction (RT-PCR).
@*RESULTS@#Compared with the Group A, the serum levels of TC and LDL-c in the Group B and Group C were significantly increased (all P<0.01). Compared with the Group B, the serum levels of TC and LDL-c in the Group C were reduced significantly after atorvastatin intervention for 4 weeks (all P<0.01). The plaques disruption and thrombosis occurred in 4 out of the 6 rabbits in the Group B, while only 1 rabbit demonstrated plaques disruption and thrombosis in the Group C. Compared with the Group B, the levels of CyPA, CD147 and macrophage in carotid atherosclerotic plaque in the Group C were decreased significantly (all P<0.01).
@*CONCLUSION@#The up-regulation of CyPA and CD147 may be involved in pathogenesis of vulnerable carotid atherosclerotic plaque. Atorvastatin could stabilize the plaque through inhibiting the CyPA and CD147 expression.


Subject(s)
Animals , Atorvastatin , Pharmacology , Basigin , Metabolism , Carotid Artery, Common , Pathology , Cholesterol , Blood , Cholesterol, Dietary , Cyclophilin A , Metabolism , Macrophages , Cell Biology , Male , Plaque, Atherosclerotic , Drug Therapy , Metabolism , Rabbits , Random Allocation , Thrombosis , Pathology , Triglycerides , Blood
4.
Chinese Medical Journal ; (24): 3191-3196, 2015.
Article in English | WPRIM | ID: wpr-275538

ABSTRACT

<p><b>BACKGROUND</b>Pelvic organ prolapse (POP) is a major health problem in adult women that involves many factors. No proteomic analysis has been conducted exclusively in POP patients. This study aimed to identify the differential expression of proteins that may be involved in POP by proteomic analysis.</p><p><b>METHODS</b>Samples of the uterosacral ligament (USL) were collected from five POP patients and five non-POP patients matched according to age, parity, and menopausal status and analyzed using two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the mRNA expression of proteins that showed differential expression in the proteomic analyses.</p><p><b>RESULTS</b>Proteins differentially expressed between POP and non-POP patients were detected. Eight proteins that were down-regulated in the POP group were identified by MALDI-TOF-MS. These proteins included electron transfer flavoprotein, apolipoprotein A-I, actin, transgelin, cofilin-1, cyclophilin A, myosin, and galectin-1, and their expression was verified by qRT-PCR.</p><p><b>CONCLUSION</b>Using comparative proteomics, we identified eight differentially expressed proteins (including four cytoskeleton proteins and three proteins related to apoptosis) in the USL that may be involved in apoptosis associated with the tissue effects in POP pathophysiology.</p>


Subject(s)
Actins , Metabolism , Aged , Apolipoprotein A-I , Metabolism , Cyclophilin A , Metabolism , Cytoskeleton , Metabolism , Female , Flavoproteins , Metabolism , Galectin 1 , Metabolism , Humans , Ligaments , Metabolism , Microfilament Proteins , Metabolism , Middle Aged , Muscle Proteins , Metabolism , Myosins , Metabolism , Pelvic Organ Prolapse , Metabolism , Postmenopause , Metabolism , Proteomics , Methods , Reverse Transcriptase Polymerase Chain Reaction , Sacrum , Metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Uterus , Metabolism
5.
Acta Physiologica Sinica ; (6): 393-400, 2015.
Article in English | WPRIM | ID: wpr-255934

ABSTRACT

The changes of serum cyclophilin A (CyPA), its receptor CD147 and the downstream signaling pathway during the process of cardiac hypertrophy remain unknown. The present study aims to investigate the relationships between CyPA-CD147-ERK1/2-cyclin D2 signaling pathway and the development of cardiac hypertrophy. Left ventricular hypertrophy was prepared by 2-kidney, 2-clip in Sprague-Dawley rats and observed for 1 week, 4 and 8 weeks. Left ventricular hypertrophy was evaluated by ratio of left ventricular heart weight to body weight (LVW/BW) and cardiomyocyte cross sectional area (CSA). CyPA levels in serum were determined with a rat CyPA ELISA kit. Expressions of CyPA, CD147, phospho-ERK1/2 and cyclin D2 in left ventricular myocytes were determined by Western blot and immunostaining. Compared with sham groups, systolic blood pressure reached hypertensive levels at 4 weeks in 2K2C groups. LVW/BW and CSA in 2K2C groups were significantly increased at 4 and 8 weeks after clipping. ELISA results indicated a prominent increase in serum CyPA level associated with the degree of left ventricular hypertrophy. Western blot revealed that the expressions of CyPA, CD147, phospho-ERK1/2 and cyclin D2 in left ventricular tissues were also remarkably increased as the cardiac hypertrophy developed. The results of the present study demonstrates that serum CyPA and CyPA-CD147-ERK1/2-cyclin D2 signaling pathway in ventricular tissues are time-dependently upregulated and activated with the process of left ventricular hypertrophy. These data suggest that CyPA-CD147 signaling cascade might play a role in the pathogenesis of left ventricular hypertrophy, and CyPA might be a prognosticator of the degree of left ventricular hypertrophy.


Subject(s)
Animals , Basigin , Metabolism , Blood Pressure , Cyclin D2 , Cyclophilin A , Metabolism , Hypertension , Hypertrophy, Left Ventricular , Metabolism , Mitogen-Activated Protein Kinase 1 , Metabolism , Mitogen-Activated Protein Kinase 3 , Metabolism , Myocytes, Cardiac , Rats , Rats, Sprague-Dawley , Signal Transduction , Up-Regulation
6.
Einstein (Säo Paulo) ; 12(3): 336-341, Jul-Sep/2014. tab, graf
Article in English | LILACS | ID: lil-723916

ABSTRACT

Objective A growing number of published articles report the expression of specific genes with different behavior patterns in rats. The levels of messenger ribonucleic acid transcripts are usually analyzed by reverse transcription followed by polymerase chain reaction and quantified after normalization with an internal control or reference gene (housekeeping gene). Nevertheless, housekeeping genes exhibit different expression in the central nervous system, depending on the physiological conditions and the area of the brain to be studied. The choice of a good internal control gene is essential for obtaining reliable results. This study evaluated the expression of three housekeeping genes (beta-actin, cyclophilin A, and ubiquitin C) in different areas of the central nervous system in rats (olfactory bulb, hippocampus, striatum, and prefrontal cortex). Methods Wistar rats (virgin females, n=6) during the diestrum period were used. Total ribonucleic acid was extracted from each region of the brain; the complementary deoxyribonucleic acid was synthesized by reverse transcription and amplified by real-time quantitative polymerase chain reaction using SYBR™ Green and primers specific for each one of the reference genes. The stability of the expression was determined using NormFinder. Results Beta-actin was the most stable gene in the hippocampus and striatum, while cyclophilin A and ubiquitin C showed greater stability in the prefrontal cortex and the olfactory bulb, respectively. Conclusion Based on our study, further studies of gene expression using rats as animal models should take into consideration these results when choosing a reliable internal control gene. .


Objetivo Um número crescente de artigos publicados relaciona a expressão de genes específicos com diferentes padrões de comportamento em ratos. Os níveis de transcritos de ácido ribonucleico mensageiro são geralmente analisados por transcrição reversa, seguida de reação em cadeia da polimerase, e quantificados após a normalização com um controle interno ou gene de referência (gene housekeeping). No entanto, os genes housekeeping exibem expressão diferencial no sistema nervoso central, dependendo das condições fisiológicas e da área do cérebro a ser estudada. A escolha de um bom gene de controle interno é essencial para a obtenção de resultados confiáveis. Este estudo avaliou a expressão de três genes housekeeping (beta-actina, ciclofilina A e ubiquitina C) em diferentes áreas do sistema nervoso central de ratos (bulbo olfatório, hipocampo, estriado e córtex pré-frontal). Métodos Foram usadas ratas Wistar (fêmeas virgens, n=6) durante o período de diestro. O ácido ribonucleico total foi extraído a partir de cada região do cérebro; o ácido desoxirribonucleico complementar foi sintetizado por transcrição reversa e amplificado por reação em cadeia da polimerase quantitativo em tempo real utilizando SYBR® Green e primers específicos para cada um dos genes de referência. A estabilidade de expressão foi determinada utilizando NormFinder. Resultados A beta-actina foi o gene mais estável no hipocampo e estriado, enquanto a ciclofilina A e a ubiquitina C apresentaram maior estabilidade no córtex pré-frontal e no bulbo olfatório, respectivamente. Conclusão Com base em nosso trabalho, estudos posteriores de expressão gênica utilizando ratos como modelos animais devem levar ...


Subject(s)
Animals , Female , Actins/genetics , Brain/physiology , Cyclophilin A/genetics , Ubiquitin C/genetics , Actins/analysis , Behavior, Animal , Cyclophilin A/analysis , Genes, Essential/physiology , Internal-External Control , Rats, Wistar , Real-Time Polymerase Chain Reaction , Reference Values , Reverse Transcription , RNA, Messenger/genetics , Ubiquitin C/analysis
7.
Chinese Journal of Cardiology ; (12): 593-597, 2014.
Article in Chinese | WPRIM | ID: wpr-316405

ABSTRACT

<p><b>OBJECTIVE</b>The aim of this study was to determine whether inhibition of cyclophilin A by lentivirus-mediated RNA interference (RNAi) could inhibit progression of atherosclerotic plaques and increase collagen production.</p><p><b>METHODS</b>Atherosclerostic plaque model was induced by rapid perivascular carotid silicone collar placement in ApoE(-/-) mice. The recombinant CyPA-RNAi-Lentivirus (CyPA-RNAi-LV) or negative control-green fluorescent protein-Lentivirus (NC-GFP-LV) were constructed and transfected into right carotid plaques, respectively. Using the local injection method, ApoE(-/-) mice carotid artery plaque were intervened 10 min in the silicone collar placement with 10 µl (1.0 × 10⁸ TU/ml) lentivirus vector. The areas and CyPA expression of plaques were analyzed by morphological observation, real-time polymerase chain reaction (RT-PCR) and Western blot respectively.</p><p><b>RESULTS</b>CyPA-RNAi-LV not only prevented plaques progression ((9 085 ± 671) µm² to (18 021 ± 1 901) µm²), but also decreased plaque lipid content ((28.9 ± 6.3)% to (17.8 ± 4.5)%), increased plaque collagen content ((24.2 ± 4.8)% to (35.1 ± 5.2)%) at 6 weeks after lentivirus transfection. The intima/media ratio (0.36 ± 0.11 vs. 0.65 ± 0.12, P < 0.05) and degree of lumen stenosis (intima/lumen ratios, 0.18 ± 0.02 vs. 0.33 ± 0.03, P < 0.05) were also significantly reduced by CyPA-RNAi-LV. Moreover, RT-PCR analysis revealed downregulated expressions of proinflammatory cytokines and matrix metalloproteinases (MMP-9 -17.5%) in the CyPA-RNAi-LV group.</p><p><b>CONCLUSION</b>Lentivirus-mediated CyPA silencing by siRNA could inhibit plaques progression and reduce local inflammation through the anti-inflammatory effects in this model.</p>


Subject(s)
Animals , Apolipoproteins E , Cyclophilin A , Genetics , Disease Progression , Gene Silencing , Genetic Vectors , Inflammation , Lentivirus , Matrix Metalloproteinase 9 , Matrix Metalloproteinases , Mice , Plaque, Atherosclerotic , RNA Interference , RNA, Small Interfering , Reverse Transcriptase Polymerase Chain Reaction , Transfection
8.
Article in English | WPRIM | ID: wpr-727507

ABSTRACT

Hepatocellular carcinoma (HCC) related to hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is thought to account for more than 80% of primary liver cancers. Both HBV and HCV can establish chronic liver inflammatory infections, altering hepatocyte and liver physiology with potential liver disease progression and HCC development. Cyclophilin A (CypA) has been identified as an essential host factor for the HCV replication by physically interacting with the HCV non structural protein NS5A that in turn interacts with RNA-dependent RNA polymerase NS5B. CypA, a cytosolic binding protein of the immunosuppressive drug cyclosporine A, is overexpressed in many cancer types and often associated with malignant transformation. Therefore, CypA can be a good target for molecular cancer therapy. Because of antiviral activity, the CypA inhibitors have been tested for the treatment of chronic hepatitis C. Nonimmunosuppressive Cyp inhibitors such as NIM811, SCY-635, and Alisporivir have attracted more interests for appropriating CypA for antiviral chemotherapeutic target on HCV infection. This review describes CypA inhibitors as a potential HCC treatment tool that is contrived by their obstructing chronic HCV infection and summarizes roles of CypA in cancer development.


Subject(s)
Carcinoma, Hepatocellular , Carrier Proteins , Cyclophilin A , Cyclophilins , Cyclosporine , Cyclosporins , Cytosol , Hepacivirus , Hepatitis B virus , Hepatitis C , Hepatitis C, Chronic , Hepatitis , Hepatocytes , Liver , Liver Diseases , Liver Neoplasms , RNA-Dependent RNA Polymerase
9.
Article in English | WPRIM | ID: wpr-218110

ABSTRACT

The effects of the an immunosuppressive drug cyclosporine A (CsA), on the salivary gland are largely unknown, even though clinical trials for the stimulation of salivation using CsA have been attempted. Cyclophilin A (CypA) is known to be a binding protein for CsA. CypA has cell proliferation and tissue matrix change activities. In our present study, the presence of CypA in the gland and effects of CsA on CypA expression were investigated by immunohistochemistry, immunoblotting and RT-PCR analyses. CypA was immunohistochemically detected in various kinds of ducts in the submandibular glands of Sprague Dawley rats. The CypA mRNA level was highest at postnatal day 1 and gradually decreased in a time-dependent manner up to adulthood. The expression of CypA increased after a 10 day subcutaneous administration of CsA in postnatal day 1 rats. Surgical sections of the chorda-lingual nerve with impaired salivation showed no changes in CypA expression. A cell proliferation assay using PCNA anti-serum showed increased cell division following CsA treatment. These results suggest that CsA and CypA may act on ductal cells to regulate saliva composition rather than salivation levels.


Subject(s)
Animals , Benzeneacetamides , Carrier Proteins , Cell Division , Cell Proliferation , Cyclophilin A , Cyclosporine , Immunoblotting , Immunohistochemistry , Piperidones , Proliferating Cell Nuclear Antigen , Rats , Rats, Sprague-Dawley , RNA, Messenger , Saliva , Salivary Glands , Salivation , Submandibular Gland
10.
Article in Chinese | WPRIM | ID: wpr-326661

ABSTRACT

<p><b>OBJECTIVE</b>To explore the action mechanism of Jiedu Quyu Zishen Recipe (JQZR) on the signal transduction of glucocorticoid receptor alpha (GRalpha) in the renal tissue of MRL/lpr mice.</p><p><b>METHODS</b>Thirty MRL/lpr mice were randomly divided into three groups, i.e., the model group, the Western medicine group, and the Chinese medicine group, 10 in each. Besides, another 10 Kunming mice was taken as the normal control group. The pathological changes of the renal tissue were observed using HE staining. The expression of GRalpha was analyzed using Real-time PCR and Western blot. The effects of JQZR on the binding power of GRalpha to cyclophilin A were detected using co-immunoprecipitation.</p><p><b>RESULTS</b>The renal injury degree of MRL/lpr mice in the Western medicine group and the Chinese medicine group was alleviated. Compared with the model group, the relative quantitation of GRalpha mRNA and protein expressions in the renal tissue of mice in the Western medicine group decreased, while they increased in the Chinese medicine group, showing statistical difference (P < 0.01). JQZR could significantly elevate the binding potency of GRalpha to cyclophilin A.</p><p><b>CONCLUSION</b>Up-regulating the expression of GRalpha and enhancing mutual actions of GRalpha and cyclophilin A was one of JQZR's effects on improving the lesion of the renal tissue.</p>


Subject(s)
Animals , Cyclophilin A , Metabolism , Drugs, Chinese Herbal , Pharmacology , Kidney , Metabolism , Pathology , Lupus Nephritis , Metabolism , Pathology , Mice , Mice, Inbred MRL lpr , Mice, Inbred Strains , Receptors, Glucocorticoid , Metabolism , Signal Transduction
11.
Article in Chinese | WPRIM | ID: wpr-748046

ABSTRACT

OBJECTIVE@#To study differential expression of MRP8, CypA protein in the patients of laryngeal squamous cell carcinoma (LSCC) and the relationship in the development of LSCC.@*METHOD@#Immunohistochemistry was used to detect the expression of MRP8,CypA protein in LSCC tissues of 41 cases and matched paraneoplastic normal tissues of 41 cases,with results compared to the clinical data to determine significance.@*RESULT@#The expression of MRP8, CypA protein in carcinoma and normal tissues and composition of different positive grades were in statistical significance (P 0.05), but related to pathological stage (P < 0.05).@*CONCLUSION@#MRP8 protein is on intimate terms with different pathological differentiation stage of LSCC. MRP8, CypA protein may play an important role in the development and progression of LSCC.


Subject(s)
Adult , Aged , Calgranulin A , Metabolism , Carcinoma, Squamous Cell , Metabolism , Pathology , Cyclophilin A , Metabolism , Female , Humans , Laryngeal Neoplasms , Metabolism , Pathology , Male , Middle Aged , Neoplasm Staging , Prognosis
12.
Article in English | WPRIM | ID: wpr-27661

ABSTRACT

Cyclophilin A (CyPA) is a 17 kDa, ubiquitously expressed multifunctional protein that possesses peptidylprolyl cis-trans isomerase activity and scaffold function. Its expression is increased in inflammatory conditions including rheumatoid arthritis, autoimmune disease and cancer. Intracellular CyPA regulates protein trafficking, signal transduction, transcription regulation and the activity of certain other proteins. Secreted CyPA activates cardiovascular cells resulting in a variety of cardiovascular diseases; including vascular remodeling, abdominal aortic aneurysms formation, atherosclerosis, cardiac hypertrophy and myocardial ischemic reperfusion injury.


Subject(s)
Aortic Aneurysm, Abdominal , Arthritis, Rheumatoid , Atherosclerosis , Autoimmune Diseases , Cardiomegaly , Cardiovascular Diseases , Cyclophilin A , Cyclophilins , Myocardial Reperfusion Injury , Oxidative Stress , Protein Transport , Proteins , Quaternary Ammonium Compounds , Signal Transduction
13.
Article in Chinese | WPRIM | ID: wpr-275772

ABSTRACT

<p><b>OBJECTIVE</b>To construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.</p><p><b>METHODS</b>First, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.</p><p><b>RESULTS</b>It was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Lentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.</p>


Subject(s)
Animals , Carcinoma, Non-Small-Cell Lung , Genetics , Cell Line, Tumor , Cyclophilin A , Genetics , Gene Silencing , Genetic Vectors , Humans , Lentivirus , Genetics , Lung Neoplasms , Genetics , Mice , RNA, Small Interfering
14.
Immune Network ; : 90-97, 2009.
Article in English | WPRIM | ID: wpr-204427

ABSTRACT

BACKGROUND: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation. METHODS: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface. RESULTS: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-kappaB. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of IkappaB, and nuclear translocation of NF-kappaB p65 and p50 subunits. CONCLUSION: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.


Subject(s)
Atherosclerosis , Cell Line , Cyclophilin A , Endothelial Cells , Humans , Inflammation , Leukocytes , Macrophages , NF-kappa B , Phosphorylation , Plaque, Atherosclerotic , Reproduction
15.
Chinese Medical Journal ; (24): 716-724, 2009.
Article in English | WPRIM | ID: wpr-279848

ABSTRACT

<p><b>BACKGROUND</b>beta-amyloid peptide (Abeta) is considered responsible for the pathogenesis of Alzheimer's disease (AD). Possible mechanisms underlying Abeta-induced neuronal cytotoxicity include excessive production of reactive oxidative species (ROS) and apoptosis. Cyclophilin A (CypA), exhibits antioxidant properties and protects neurons against oxidative stress induced injury. This study was conducted to demonstrate whether CyPA added to cultured PC12 cells could alleviate Abeta-induced oxidative stress and protect them from apoptosis.</p><p><b>METHODS</b>PC12 cells were pre-incubated for 30 minutes with recombinant human cyclophilin A (rhCyPA) in 0.1 nmol/L, 1.0 nmol/L, 10 nmol/L and 100 nmol/L and then incubated with 10 micromol/L Abeta(25-35). In every group, cell viability, apoptotic morphology, apoptotic rate, intracellular ROS accumulation, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of PC12 cells and mitochondrial transmembrane potential were detected. Subsequently, the expression of the active form of caspase-3 was determined by Western blotting.</p><p><b>RESULTS</b>It was shown that cultures treated with 1.0 nmol/L, 10 nmol/L or 100 nmol/L rhCyPA + Abeta(25-35) had significantly higher cell viability and a lower rate of apoptosis compared with the cultures exposed only to Abeta(25-35). In addition, rhCyPA attenuated Abeta(25-35)-induced overproduction of intracellular ROS and Abeta(25-35)-induced a decrease in activity of the key antioxidant enzymes SOD and GSH-Px. Furthermore, rhCyPA also attenuated Abeta(25-35)-induced mitochondrial dysfunction and the activation of caspase-3.</p><p><b>CONCLUSION</b>CyPA may act as an ROS scavenger, and prevent Abeta(25-35)-induced neurotoxicity through attenuating oxidative stress induced by Abeta(25-35).</p>


Subject(s)
Amyloid beta-Peptides , Pharmacology , Animals , Caspase 3 , Metabolism , Cyclophilin A , Pharmacology , Glutathione Peroxidase , Metabolism , Humans , Oxidative Stress , PC12 Cells , Peptide Fragments , Pharmacology , Rats , Superoxide Dismutase , Metabolism
16.
Indian J Biochem Biophys ; 2008 Dec; 45(6): 374-8
Article in English | IMSEAR | ID: sea-27413

ABSTRACT

The expression of glutathione-S-transferase (GST) fusion protein is extensively utilized in the study of protein-protein interactions. In the commonly used purification method, the overexpressed GST fusion protein is bound to the glutathione (GSH)-coupled resins via affinity chromatography, and then eluted by an excessive quantity of reduced GSH. However, this technique has certain limitations, such as low product purity, retention of GSH in the sample, as well as relatively high cost. To overcome these limitations, in this study, elution buffer containing 2% formic acid was utilized rather than GSH to elute the GST-fusion protein, and thereafter the acidic samples were neutralized using collecting buffer. By using this method, highly purified GST-cyclophilin A (CypA) fusion protein was obtained, without affecting the structural and functional characteristics such as PPIase and chaperone activities. Moreover, the procedure is also cost-effective, due to the low cost of formic acid as compared with GSH.


Subject(s)
Animals , Cloning, Molecular , Cyclophilin A/genetics , Escherichia coli/enzymology , Formates/chemistry , Glutathione Transferase/genetics , Molecular Chaperones/genetics , Protein Binding , Protein Folding , Rats , Recombinant Fusion Proteins/genetics
17.
Chinese Journal of Hematology ; (12): 793-796, 2008.
Article in Chinese | WPRIM | ID: wpr-239959

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of CD147 in the proliferation, activation and chemotaxis of Jurkat cell induced by cyclophilin A (CyPA).</p><p><b>METHODS</b>CD147 mRNA and protein level siRNA transfected Jurkat cells were identified by RT-PCR and Western blot respectively. Jurkat cell, Jurkat-vector cell and Jurkat-CD147 siRNA cells were treated with different concentrations of CyPA (0.01 nmol/L, 0.1 nmol/L, 1 nmol/L, 10 nmol/L) or PBS for 24 h and 48 h. Proliferation level was detected by MTT assay. CD25 was measured by flow cytometry. Transwell chamber was used to detect the chemotaxis. The effect of CyPA on the adhesive potential of Jurkat cell was studied by cell-matrix adhesion assay.</p><p><b>RESULTS</b>CD147 mRNA and protein level siRNA transfected cells were decreased significantly than that of control cells. CyPA stimulated the proliferation of Jurkat cell in a dose-dependent manner, its effect peaked at 10 nmol/L CyPA. Blockage of CD147 expression decreased the proliferation level of Jurkat cell induced by CyPA. CyPA increased the activation rate of Jurkat cell, and blockage CD147 expression decreased the activation rate of the cell induced by CypA. CyPA showed a chemotactic activity on Jurkat cell, the chemotaxis index being 2.32, and the chemotactic ability was decreased after inhibition of CD147 expression. CyPA had no effect on adhesion of Jurkat cell to extracellular matrix.</p><p><b>CONCLUSION</b>CD147 plays a role in the proliferation, activation and chemotaxis of Jurkat cell induced by CyPA.</p>


Subject(s)
Basigin , Genetics , Cell Adhesion , Cell Proliferation , Chemotaxis , Cyclophilin A , Pharmacology , Humans , Interleukin-2 Receptor alpha Subunit , Metabolism , Jurkat Cells , RNA, Small Interfering , Genetics , Transfection
SELECTION OF CITATIONS
SEARCH DETAIL