Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 65
Filter
1.
Acta Physiologica Sinica ; (6): 772-780, 2021.
Article in Chinese | WPRIM | ID: wpr-921280

ABSTRACT

The development of nonalcoholic fatty liver disease (NAFLD) is closely related to the fatty acid (FA) uptake. This study aimed to investigate the effect of Krüppel-like factor 9 (KLF9) on CD36 (typical fatty acid translocase), hepatocellular lipid metabolism as well as the development and progression of nonalcoholic fatty liver. High-fat diet-induced obese C57BL/6J mice and db/db mice were used to test the expression levels of Klf9 and Cd36 in the livers. The primary hepatocytes were isolated from C57BL/6J mice, treated with Ad-GFP, Ad-Klf9, Ad-shCtrl or Ad-shKlf9, and then incubated with oleic acid and palmitic acid for 24 h. Liver-specific knockout of Klf9 mice were established. The protein levels and relative mRNA levels were examined by Western blot and real-time PCR, respectively. Triglyceride content was determined by using an assay kit. Lipid content was determined by Oil Red O staining. The results showed that: (1) Klf9 expression levels were increased in the livers of high-fat diet-induced obese mice and db/db mice, compared to their respective control mice. (2) Adenovirus-mediated overexpression of Klf9 in primary hepatocytes increased Cd36 expression and cellular triglyceride contents. (3) In contrast, adenovirus-mediated knockdown of Klf9 expression in primary hepatocytes by Ad-shKlf9 decreased Cd36 expression and cellular triglyceride contents. (4) Finally, Klf9 deficiency decreased liver Cd36 expression and alleviated fatty liver phenotype of high-fat diet-induced obese mice. These results suggest that KLF9 can regulate hepatic lipid metabolism and development of NAFLD by promoting the expression of CD36.


Subject(s)
Animals , CD36 Antigens/metabolism , Diet, High-Fat , Kruppel-Like Transcription Factors/metabolism , Lipid Metabolism , Liver , Mice , Mice, Inbred C57BL , Mice, Knockout , Non-alcoholic Fatty Liver Disease/metabolism , Oleic Acid/metabolism
2.
Journal of Experimental Hematology ; (6): 1463-1468, 2019.
Article in Chinese | WPRIM | ID: wpr-775698

ABSTRACT

OBJECTIVE@#To investigate the transcriptional regulation of transcription factor MZF-1 on acute monocytic leukemia-related gene MLAA-34.@*METHODS@#The effect of MZF-1 on the transcriptional activity of MLAA-34 gene promoter was analyzed by luciferase reporter gene detection system and site-directed mutation technique. The EMSA and ChIP assay were used to verify whether MZF-1 directly and specifically binds to the core region of MLAA-34 promoter. The over-expression vector and interference vector of MZF-1 were constructed to transfect U937 cells, and RT-PCR and Western blot were used to detect the transcription and expression changes of MLAA-34 gene.@*RESULTS@#The transcription factor MZF-1 had a regulatory effect on MLAA-34 gene expression, and the relative luciferase activity was decreased after MZF-1 binding point mutation (P<0.01). EMSA and ChIP experiments demonstrated that MZF-1 could directly bind to MLAA-34 promoter and play a regulatory role. In the over-expression test, the increase of MZF-1 could up-regulate the expression of MLAA-34 (P<0.05). In the interference test, the decrease of MZF-1 could down-regulate the expression of MLAA-34 (P<0.05).@*CONCLUSION@#Transcription factor MZF-1 can bind to the transcriptional regulatory region on the promoter of MLAA-34 gene and promote the transcription of MLAA-34 gene in acute monocytic leukemia.


Subject(s)
Antigens, Neoplasm , Genetics , Apoptosis Regulatory Proteins , Genetics , Gene Expression Regulation, Neoplastic , Genes, Reporter , Hepatocyte Nuclear Factor 1-alpha , Humans , Kruppel-Like Transcription Factors , Metabolism , Leukemia, Monocytic, Acute , Promoter Regions, Genetic , Transcription, Genetic
3.
Neuroscience Bulletin ; (6): 419-437, 2018.
Article in English | WPRIM | ID: wpr-777045

ABSTRACT

A previous study has indicated that Krüppel-like factor 7 (KLF7), a transcription factor that stimulates Schwann cell (SC) proliferation and axonal regeneration after peripheral nerve injury, is a promising therapeutic transcription factor in nerve injury. We aimed to identify whether inhibition of microRNA-146b (miR-146b) affected SC proliferation, migration, and myelinated axon regeneration following sciatic nerve injury by regulating its direct target KLF7. SCs were transfected with miRNA lentivirus, miRNA inhibitor lentivirus, or KLF7 siRNA lentivirus in vitro. The expression of miR146b and KLF7, as well as SC proliferation and migration, were subsequently evaluated. In vivo, an acellular nerve allograft (ANA) followed by injection of GFP control vector or a lentiviral vector encoding an miR-146b inhibitor was used to assess the repair potential in a model of sciatic nerve gap. miR-146b directly targeted KLF7 by binding to the 3'-UTR, suppressing KLF7. Up-regulation of miR-146b and KLF7 knockdown significantly reduced the proliferation and migration of SCs, whereas silencing miR-146b resulted in increased proliferation and migration. KLF7 protein was localized in SCs in which miR-146b was expressed in vivo. Similarly, 4 weeks after the ANA, anti-miR-146b increased KLF7 and its target gene nerve growth factor cascade, promoting axonal outgrowth. Closer analysis revealed improved nerve conduction and sciatic function index score, and enhanced expression of neurofilaments, P0 (anti-peripheral myelin), and myelinated axon regeneration. Our findings provide new insight into the regulation of KLF7 by miR-146b during peripheral nerve regeneration and suggest a potential therapeutic strategy for peripheral nerve injury.


Subject(s)
Animals , Cell Movement , Genetics , Cell Proliferation , Genetics , Disease Models, Animal , Female , Ganglia, Spinal , Cell Biology , Gene Expression Regulation , Genetics , Physiology , HEK293 Cells , Humans , Kruppel-Like Transcription Factors , Genetics , Metabolism , Male , MicroRNAs , Genetics , Metabolism , Motor Endplate , Genetics , Myelin P0 Protein , Metabolism , Nerve Regeneration , Genetics , Physiology , Nerve Tissue Proteins , Metabolism , RNA, Small Interfering , Genetics , Metabolism , Rats , Rats, Sprague-Dawley , Rats, Wistar , Sciatic Neuropathy , Metabolism , General Surgery , Therapeutics
4.
Neuroscience Bulletin ; (6): 507-516, 2018.
Article in English | WPRIM | ID: wpr-777029

ABSTRACT

The ZNF804A variant rs1344706 has consistently been associated with schizophrenia and plays a role in hippocampal-prefrontal functional connectivity during working memory. Whether the effect exists in the resting state and in patients with schizophrenia remains unclear. In this study, we investigated the ZNF804A polymorphism at rs1344706 in 92 schizophrenic patients and 99 healthy controls of Han Chinese descent, and used resting-state functional magnetic resonance imaging to explore the functional connectivity in the participants. We found a significant main effect of genotype on the resting-state functional connectivity (RSFC) between the hippocampus and the dorsolateral prefrontal cortex (DLPFC) in both schizophrenic patients and healthy controls. The homozygous ZNF804A rs1344706 genotype (AA) conferred a high risk of schizophrenia, and also exhibited significantly decreased resting functional coupling between the left hippocampus and right DLPFC (F(2,165) = 13.43, P < 0.001). The RSFC strength was also correlated with cognitive performance and the severity of psychosis in schizophrenia. The current findings identified the neural impact of the ZNF804A rs1344706 on hippocampal-prefrontal RSFC associated with schizophrenia.


Subject(s)
Adult , Analysis of Variance , Female , Functional Laterality , Genetics , Genotype , Hippocampus , Diagnostic Imaging , Humans , Image Processing, Computer-Assisted , Kruppel-Like Transcription Factors , Genetics , Magnetic Resonance Imaging , Male , Neural Pathways , Diagnostic Imaging , Neuropsychological Tests , Oxygen , Blood , Polymorphism, Single Nucleotide , Genetics , Prefrontal Cortex , Diagnostic Imaging , Psychiatric Status Rating Scales , Schizophrenia , Diagnostic Imaging , Genetics , Severity of Illness Index , Young Adult
5.
Int. j. morphol ; 35(1): 259-264, Mar. 2017. ilus
Article in English | LILACS | ID: biblio-840964

ABSTRACT

Kruppel-like factor 6 (KLF6) is a member of the family of Kruppel transcription factors, this plays an important role in the regulation of cell growth, differentiation and angiogenesis. Rosiglitazone is a PPARy agonist drug, its antitumor effect has been described in models of breast and colon cancer. The aim of this study is to evaluate the level of expression of KLF6 in Caco2 cells treated with Avandia. For this a Immunofluorescence was performed, the Caco2 cells were cultured and treated with Rosiglitazone, another group was treated with Rosiglitazone and GW-9662, inhibitor for Immunofluorescence an anti-KLF6 antibody and a secondary antibody coupled to Alexa-488 was used . Cells were observed in a fluorescence microscope and images were processed. The results show that KLF6 is expressed in the cytoplasm of cells Caco2. Compared to treatment with Avandia, KLF6 increases its expression in the cytoplasm. When cells were treated with GW-9662 inhibitor, an expression of KLF6 in the nucleus was observed. KLF6 expression in the cytoplasm of cells Caco2, could be explained by the knowledge of splicing variants SV1 and SV2, these abnormally accumulate in the cytoplasm and promotes cell growth. It is concluded that in untreated Caco 2 cells, KLF6 is expressed in the cytoplasm. Compared to treatment with Rosiglitazone, KLF6 upregulated in the cytoplasm and compared to treatment with the inhibitor, KLF6 is expressed in the nucleus of Caco 2 cells.


Kruppel-like factor 6 (KLF6) is a member of the family of Kruppel transcription factors, this plays an important role in the regulation of cell growth, differentiation and angiogenesis. Rosiglitazone is a PPARy agonist drug, its antitumor effect has been described in models of breast and colon cancer. The aim of this study is to evaluate the level of expression of KLF6 in Caco2 cells treated with Avandia. For this a Immunofluorescence was performed, the Caco2 cells were cultured and treated with Rosiglitazone, another group was treated with Rosiglitazone and GW-9662, inhibitor for Immunofluorescence an anti-KLF6 antibody and a secondary antibody coupled to Alexa-488 was used . Cells were observed in a fluorescence microscope and images were processed. The results show that KLF6 is expressed in the cytoplasm of cells Caco2. Compared to treatment with Avandia, KLF6 increases its expression in the cytoplasm. When cells were treated with GW-9662 inhibitor, an expression of KLF6 in the nucleus was observed. KLF6 expression in the cytoplasm of cells Caco2, could be explained by the knowledge of splicing variants SV1 and SV2, these abnormally accumulate in the cytoplasm and promotes cell growth. It is concluded that in untreated Caco 2 cells, KLF6 is expressed in the cytoplasm. Compared to treatment with Rosiglitazone, KLF6 upregulated in the cytoplasm and compared to treatment with the inhibitor, KLF6 is expressed in the nucleus of Caco 2 cells.


Subject(s)
Humans , Adenocarcinoma/metabolism , Antineoplastic Agents/pharmacology , Colonic Neoplasms/metabolism , Kruppel-Like Transcription Factors , Thiazolidinediones/pharmacology , Apoptosis , Caco-2 Cells , Cell Line
6.
Medicina (B.Aires) ; 76(4): 213-218, Aug. 2016. graf, tab
Article in English | LILACS | ID: biblio-841579

ABSTRACT

Several heterozygous GLI2 gene mutations have been reported in patients with isolated GH deficiency (IGHD) or multiple pituitary hormone deficiency (MPHD) with or without other malformations. The primary aim of this study was to analyze the presence of GLI2 gene alterations in a cohort of patients with IGHD or MPHD and ectopic/absent posterior pituitary. The coding sequence and flanking intronic regions of GLI2 gene were amplified and directly sequenced from gDNA of 18 affected subjects and relatives. In silico tools were applied to identify the functional impact of newly found variants (Polyphen2, SIFT, Mutation Taster). We identified two novel heterozygous missense variations in two unrelated patients, p.Arg231Gln and p.Arg226Leu, located in the repressor domain of the protein. Both variations affect highly conserved amino acids of the Gli2 protein and were not found in the available databases. In silico tools suggest that these variations might be disease causing. Our study suggests that the GLI2 gene may be one of the candidate genes to analyze when an association of pituitary hormone deficiency and developmental defects in posterior pituitary gland. The highly variable phenotype found suggests the presence of additional unknown factors that could contribute to the phenotype observed in these patients.


Mutaciones heterocigotas en el gen GLI2 fueron previamente comunicadas como causa de déficit aislado de hormona de crecimiento (IGHD) o déficit múltiple de hormonas hipofisarias (MPHD), con o sin otras malformaciones. El objetivo del estudio fue analizar la presencia de alteraciones en el gen GLI2 en un grupo de pacientes con IGHD o MPHD acompañado de neurohipófisis ectópica o ausente. La secuencia codificante y las regiones intrónicas flanqueantes del gen GLI2 fueron amplificadas y secuenciadas de manera directa a partir de ADN genómico extraído de sangre periférica proveniente de 18 sujetos afectados y sus familiares. Se utilizaron herramientas informáticas para predecir el impacto funcional de las nuevas variantes encontradas (Polyphen2, SIFT, Mutation Taster). Identificamos dos nuevas variantes heterocigotas con pérdida de sentido en dos pacientes no relacionados, p.Arg231Gln y p.Arg226Leu, localizadas en el dominio represor de la proteína. Estas variantes afectan aminoácidos altamente conservados en la secuencia proteica de GLI2 y no se encuentran informadas en las bases de datos disponibles. Las herramientas informáticas utilizadas sugieren que estas variantes pueden ser la causa del desarrollo de la enfermedad. Nuestro resultados indican que el gen GLI2 es uno de los genes candidatos a estudiar cuando existe una asociación entre déficit de hormonas hipofisarias y alteraciones en el desarrollo de la neurohipófisis. Se sugiere la existencia de otros factores adicionales que podrían contribuir a la variabilidad del fenotipo observado.


Subject(s)
Humans , Male , Infant, Newborn , Infant , Child, Preschool , Child , Pituitary Hormones/deficiency , Human Growth Hormone/deficiency , Mutation, Missense , Kruppel-Like Transcription Factors/genetics , Phenotype , Argentina , Pituitary Gland, Anterior/abnormalities , Pituitary Gland, Posterior/abnormalities , Introns , Zinc Finger Protein Gli2 , Heterozygote , Microcephaly/diagnosis
7.
Acta Physiologica Sinica ; (6): 809-815, 2016.
Article in Chinese | WPRIM | ID: wpr-331600

ABSTRACT

Krüppel-like factor 7 (KLF7), a member of Krüppel-like transcription factors (KLFs), also known as ubiquitous Krüppel- like factor (UKLF), is ubiquitously expressed in various tissues of adult human beings. Genetics reports showed that the genetic polymorphism of KLF7 is associated with obesity, type 2 diabetes, mental development in human beings; and KLF7 methylation is associated with the development of diffuse gastric cancer (gastric adenocarcinoma). In addition, some genomics reports suggested that KLF7 is one of the key transcription factors in the regulatory networks of serum markers change during the cardiovascular disease. The function studies showed that KLF7 is involved in the regulation of the development and function of the nervous system and adipose tissue, type 2 diabetes, blood diseases, as well as pluripotent cells maintenance. This review summarizes the research progress of KLF7 in genetic characteristics, protein structure and gene function.


Subject(s)
Diabetes Mellitus, Type 2 , Humans , Kruppel-Like Transcription Factors , Metabolism , Nervous System , Obesity
8.
Article in English | WPRIM | ID: wpr-296563

ABSTRACT

We used Smo siRNA to inhibit hedgehog signaling pathway in embryonic day (E) 13 palatal shelves in organ culture. SiRNA 4 was chosen as the most efficient from four synthesized Smo siRNAs. Palatal shelf fusion rate of 4 μg/mL cyclopamine group was the lowest and significantly lower than that of blank control group (P<0.05), and that of siRNA 4 group was also lower than that of blank control group (P=0.183). At 48 h after transfection, Smo protein level of siRNA 4 group was 64.8% lower than that of blank control group (P<0.05), and Gli1 protein level of 4 μg/mL cyclopamine group was 68.9% lower than that of blank control group (P<0.05). Hedgehog signaling pathway inhibition decreased palatal fusion in organ culture, probably owing to downregulation of Smo and Gli1 proteins.


Subject(s)
Animals , Hedgehog Proteins , Genetics , Metabolism , Kruppel-Like Transcription Factors , Genetics , Metabolism , Mice , Nerve Tissue Proteins , Genetics , Metabolism , Palate , Embryology , Metabolism , RNA, Small Interfering , Genetics , Metabolism , Signal Transduction , Zinc Finger Protein Gli2 , Zinc Finger Protein Gli3
9.
Article in English | WPRIM | ID: wpr-238418

ABSTRACT

The relationship between Kruppel-like factor 4 (KLF4) and the Notch pathway was determined to investigate the effect of KLF4 on the activation of hepatic stellate cells and underlying mechanisms. Fifty SPF BALB/c mice were randomly divided into two groups. A liver fibrosis model was established in 25 mice as the experimental group, and the remaining 25 mice served as controls. On the day 0, 7, 14, and 35, liver tissues were removed for immunofluorescent detection. The Notch pathway inhibitor DAPT was added to the primary original hepatic stellate cells, and KLF4 and Notch-associated factor expression was detected by qRT-PCR. Additionally, the hepatic stellate cell line LX-2 was used to establish control and experimental groups, and was cultured in vitro. LX-2 cells in the experimental groups were treated with DAPT and the Notch activator transforming growth factor-beta 1 separately, whereas those in the control group were given isotonic culture medium. After 48 h, KLF4 expression was examined by Western blotting. After transient transfection of LX-2 cells to increase KLF4, the expression of Notch factor was examined. Immunofluorescence analysis showed that, with the aggravation of liver fibrosis, the absorbance (A) values of KLF4 were decreased (day 0: 980.73±153.19; day 7: 1087.99±230.23; day 14: 390.95±93.56; day 35: 245.99±87.34). The expression of Notch pathway- related factors (Notch-1, Notch-2, and Jagged-1) in the hepatic stellate cell membrane was negatively correlated to KLF4 expression. With the increase of KLF4 expression, Notch-2 (0.73±0.13) and Jagged-1 (0.43±0.12) expression decreased, whereas Notch-1 level was not detectable. When the Notch pathway was inhibited, KLF4 levels generally increased (18.12±1.31). Our results indicate that KLF4 expression is negatively correlated to the Notch pathway in hepatic stellate cells, which may provide a reference for the treatment of hepatic fibrosis.


Subject(s)
Animals , Cell Line , Cells, Cultured , Hepatic Stellate Cells , Metabolism , Kruppel-Like Transcription Factors , Genetics , Metabolism , Liver Cirrhosis , Metabolism , Mice , Mice, Inbred BALB C , Receptors, Notch , Metabolism , Signal Transduction , Transforming Growth Factor beta1 , Metabolism
10.
Article in Chinese | WPRIM | ID: wpr-279891

ABSTRACT

Macrophages have two major roles in regulating the dynamic equilibrium in erythropoiesis, promoting the differentiation and maturation of nucleated red blood cells into reticulocytes and removing old red blood cells. A recent mouse study has demonstrated that the phenotype of macrophages in erythroblastic islands is CD169+ VCAM-1+ ER-HR3+ CD11b+ F4/80+ Ly-6G+. Molecular connections between erythroid progenitor cells and central macrophages help to maintain the function and integrity of erythroblastic islands. New research advances in Kruppel-like factor 1 (KLF1) provide new evidence for the important role of macrophages in erythroblastic islands. Macrophages play an important role in erythropoiesis both in sickness and in health, and provide a potential targeted therapy for diseases such as polycythemia vera and beta-thalassemia in the future.


Subject(s)
Animals , Erythropoiesis , Humans , Integrin alpha4beta1 , Physiology , Kruppel-Like Transcription Factors , Physiology , Macrophages , Physiology , Mice , Phenotype , Vascular Cell Adhesion Molecule-1 , Physiology
11.
Chinese Journal of Cardiology ; (12): 162-166, 2015.
Article in Chinese | WPRIM | ID: wpr-328818

ABSTRACT

<p><b>OBJECTIVE</b>To explore the impact of kruppel like factor 15 (KLF15) on cardiac fibroblasts on angiogenesis in a pressure overload rat model.</p><p><b>METHODS</b>Pressure overload was induced in female rats by aortic constriction for 3 and 6 weeks. After 6 weeks aortic banding, rats underwent aortic debanding for 3 or 6 weeks. Sham rats were observed for 3 and 6 weeks (n = 10 each). Cardiac function, myocardial pathological changes, interstitial angiogenesis and KLF15 expression during rat myocardial overloading-unloading process were determined. Cardiac fibroblasts and vascular endothelial cells were cultured in vitro in the absence or presence of KLF15-shRNA recombinant adenovirus and the regulation effect of KLF15 on vascular endothelial cells and angiogenesis was observed on a three-dimensional angiogenesis in vitro model.</p><p><b>RESULTS</b>The ascending aorta diameter, ejection fraction, fractional shortening, left ventricular systolic pressure and the KLF15 protein expression level were significantly lower but the left ventricular end-diastolic pressure was significantly higher in pressure overloaded rats than in Sham rats (all P < 0.01) after 6 weeks. At the same time, increased myocardial hypertrophy and fibrosis as well as reduced angiogenesis density were observed in pressure overloaded rats. These changes were significantly attenuated post aortic debanding. In vitro, KLF15-shRNA recombinant adenovirus transfection into cardiac fibroblasts significantly downregulated the protein expression of KLF15 compared with the control group (4 922 ± 430 vs. 7 034 ± 178, P < 0.01). The formation of tubular structure of vascular endothelial cells was shorter after KLF15-shRNA recombinant adenovirus transfection and the structure was incomplete when compared with the control group.</p><p><b>CONCLUSION</b>Our results suggest that upregulation of KLF15 expression in myocardial fibroblasts might promote vascular generation, alleviate the myocardial interstitial fibrosis and improve cardiac function in this pressure overload rat model.</p>


Subject(s)
Animals , Aorta , Aortic Valve Stenosis , Down-Regulation , Female , Fibrosis , Heart , Kruppel-Like Transcription Factors , Metabolism , Myocardium , Neovascularization, Physiologic , Rats , Vascular Remodeling
12.
Article in Chinese | WPRIM | ID: wpr-239177

ABSTRACT

<p><b>OBJECTIVE</b>To screen and identify the proteins related with tumor metastasis of gastric cancer in a nude mouse model bearing orthotopic transplanted tumor.</p><p><b>METHODS</b>Zinc finger protein 139 (ZNF139)-specific siRNA was synthesized and transfected into gastric cancer cell line SGC7901, which was then screened by G418. ZNF139-siRNA-transfected cells, negative plasmid-transfected cells and untreated SGC7901 cells were orthotopically transplanted separately on the stomach wall of BALB/c nude mice. The primary tumors and metastatic lymph nodes were harvested to separate the proteins by 2-D fluorescence difference gel electrophoresis (2-D DIGE); after gel digestion, the differential proteins were subjected to liquid chromatography-mass spectrometry (LC-MS) for identification and their functions were analyzed. Western blotting was performed to verify the identified proteins.</p><p><b>RESULTS</b>ZNF139 expression was effectively inhibited in siRNA-transfected SGC7901 cells. ZNF139-siRNA-transfected cells showed obviously suppressed tumor growth with a lowered lymph node metastasis rate in nude mice compared with untreated cells and the negative control cells (P<0.05). Proteomic study with 2-D DIGE showed that fascin and hnRNPA2/B1 were down-regulated while ANXA1 was up-regulated in the primary tumors, and ANXA5 was down-regulated in the metastatic lymph nodes in ZNF139-siRNA-transfected group. Western blotting confirmed the results of proteomic analysis.</p><p><b>CONCLUSION</b>ZNF139 gene may promote lymph node metastasis of gastric cancer by regulating fascin, hnRNPA2/B1, ANXA1, and ANXA5.</p>


Subject(s)
Animals , Annexins , Metabolism , Blotting, Western , Cell Line, Tumor , Chromatography, Liquid , Electrophoresis, Gel, Two-Dimensional , Heterogeneous-Nuclear Ribonucleoprotein Group A-B , Metabolism , Humans , Kruppel-Like Transcription Factors , Metabolism , Lymphatic Metastasis , Mice , Mice, Nude , Neoplasm Proteins , Metabolism , Neoplasm Transplantation , Proteomics , RNA, Small Interfering , Stomach Neoplasms , Pathology , Transfection
13.
Article in English | WPRIM | ID: wpr-264613

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of bisdemethoxycurcumin (BDMC) on non-small cell lung cancer (NSCLC) cell line, A549, and the highly metastatic lung cancer 95D cells.</p><p><b>METHODS</b>CCK-8 assay was used to assess the effect of BDMC on cytotoxicity. Flow cytometry was used to evaluate apoptosis. Western blot analysis, electron microscopy, and quantification of GFP-LC3 punctuates were used to test the effect of BDMC on autophagy and apoptosis of lung cancer cells.</p><p><b>RESULTS</b>BDMC inhibited the viability of NSCLC cells, but had no cytotoxic effects on lung small airway epithelial cells (SAECs). The apoptotic cell death induced by BDMC was accompanied with the induction of autophagy in NSCLC cells. Blockage of autophagy by the autophagy inhibitor 3-methyladenine (3-MA) repressed the growth inhibitory effects and induction of apoptosis by BDMC. In addition, BDMC treatment significantly decreased smoothened (SMO) and the transcription factor glioma-associated oncogene 1 (Gli1) expression. Furthermore, depletion of Gli1 by siRNA and cyclopamine (a specific SMO inhibitor) induced autophagy.</p><p><b>CONCLUSION</b>Aberrant activation of Hedgehog (Hh) signaling has been implicated in several human cancers, including lung cancers. The present findings provide direct evidence that BDMC-induced autophagy plays a pro-death role in NSCLC, in part, by inhibiting Hedgehog signaling.</p>


Subject(s)
Antineoplastic Agents , Pharmacology , Apoptosis , Autophagy , Carcinoma, Non-Small-Cell Lung , Drug Therapy , Cell Line, Tumor , Curcumin , Chemistry , Pharmacology , Down-Regulation , Gene Expression Regulation, Neoplastic , Hedgehog Proteins , Genetics , Metabolism , Humans , Kruppel-Like Transcription Factors , Genetics , Metabolism , Signal Transduction , Zinc Finger Protein GLI1
14.
Article in English | WPRIM | ID: wpr-76671

ABSTRACT

BACKGROUND/AIMS: A number of genome-wide and candidate gene association studies have identified polymorphisms associated with telomere length in Caucasian populations. This study was conducted to determine the impacts of 17 polymorphisms identified in Caucasians on telomere length in a Korean population. METHODS: Ninety-four healthy individuals were enrolled in this study. Relative telomere length of chromosomes from peripheral blood samples was measured using quantitative polymerase chain reaction. RESULTS: Two polymorphisms, rs10936599 of MYNN and rs412658 of ZNF676, were found to be associated w ith telomere length (under dominant model, p = 0.04; under recessive model, p = 0.001). Three polymorphisms, rs2853669, rs7705526, and rs2736108, at the TERT locus were also associated with telomere length (under recessive model, p = 0.01, p = 0.02, and p = 0.01, respectively). The genotypes of the five polymorphisms associated with short telomere length were considered bad genotypes; telomere length was significantly decreased with increasing number of bad genotypes (p= 1.7 x 10(-5)). CONCLUSIONS: We have identified polymorphisms associated with telomere length in a Korean population.


Subject(s)
Adult , Aged , Asians/genetics , Case-Control Studies , DNA-Binding Proteins/genetics , Female , Genome-Wide Association Study , Genotype , Humans , Kruppel-Like Transcription Factors/genetics , Male , Middle Aged , Phenotype , Polymorphism, Single Nucleotide , Republic of Korea , Telomerase/genetics , Telomere/genetics , Telomere Homeostasis , Zinc Fingers
15.
Chinese Journal of Pathology ; (12): 814-819, 2014.
Article in Chinese | WPRIM | ID: wpr-304383

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of sonic hedgehog (Shh) signaling pathway in liver fluorosis and to explore related mechanism.</p><p><b>METHODS</b>To establish animal model, 48 normal SD rats (aged 4-5 weeks) were randomly divided into 4 groups (12 each): control group, fluoriosis group, blocking group and blocking control group. After 6 months, the blocking group and blocking control group were injected intraperitoneally once every 2 days for 3 times with 10 mg/kg cyclopamine or dimethysulfoxide, respectively. Rats were sacrificed at the end of the experiment and the fluoride content in urine and liver function was determined. The expression of Shh and Gli1 protein and mRNA in hepatocytes was detected by immunohistochemistry and real-time fluorescence quantitative PCR, respectively.</p><p><b>RESULTS</b>The fluoride contents in the urine and the incidence of dental fluorosis increased in the fluoride and blocking control groups as compared with those in the control group, but decreased in the blocking group compared with those of the fluoride and blocking control group. Compared with the control group, the titers of aspartate transaminase (AST) and alanine transaminase (ALT) significantly increased, while the activity of total protein and albumin decreased in the fluoride and blocking control groups. Compared with the fluoride and blocking control groups, the activity of the ALT slightly declined and the AST, total protein and albumin slightly increased in the blocking group. Histologically, the cells were disorganized and swollen with cytoplasmic clearing (balloon cells), compared with the control group. The expression of Shh and Gli1 significantly increased in all but the control group. Compared with the fluoride and blocking control groups, the expression of Shh and Gli1 declined in the blocking group.</p><p><b>CONCLUSIONS</b>The overexpression and cyclopamine inhibition of the Shh signaling pathway are closely related to the content of fluoride in the liver. The Shh signaling pathway plays an important role in the pathogenesis of liver injury caused by fluorosis, suggesting a preventive and therapeutic target of the disease.</p>


Subject(s)
Alanine Transaminase , Animals , Aspartate Aminotransferases , Dimethyl Sulfoxide , Pharmacology , Disease Models, Animal , Fluoride Poisoning , Drug Therapy , Metabolism , Fluorosis, Dental , Diagnosis , Hedgehog Proteins , Metabolism , Hepatocytes , Metabolism , Kruppel-Like Transcription Factors , Metabolism , Liver , Metabolism , Liver Diseases , Drug Therapy , Metabolism , RNA, Messenger , Random Allocation , Rats , Rats, Sprague-Dawley , Signal Transduction , Veratrum Alkaloids , Pharmacology , Zinc Finger Protein GLI1
16.
Journal of Experimental Hematology ; (6): 1278-1281, 2014.
Article in Chinese | WPRIM | ID: wpr-340514

ABSTRACT

The aim of this study was to investigate the effect of TIEG1 on K562 cell apoptosis and expression of BCL-2/BAX, PTEN. The different concentration(0, 1, 5, 10, 20 ng/ml) of TIEG1 were used to treat K562 cells, the cell growth inhibition rate was detected by using MTT method. After treating K562 cells with 10.00 ng/ml TIEG1, the cell apoptosis was detected with flow cytometry. The RT-PCR was used to detected the expression levels of BCL-2 /BAX and PTEN. The results showed that TIEG1 displays inhibitory effect on proliferation of K562 cells in time-and dose-dependent manner (r = 0.52, P < 0.05) ; after K562 cells were treated for 6, 12, 24 and 48 h, the IC50 of TIEG1 were 48.19, 18.72, 9.5 and 3.85 ng/ml respectively. After treating K562 cells with 10.00 ng/ml TIEG1 for 0, 6, 12, 24, 48 h, the apoptosis rate were (2.13 ± 0.42)%, (7.79 ± 0.71)%, (11.17 ± 1.37)%, (24.66 ± 0.29)% and (48.60 ± 1.38)% respectively, and there was significant difference between groups(P < 0.05). In process of K562 cell apoptosis, the expression level of BCL-2 gradually decreased (r = 0.48, P < 0.05), meanwhile the expression levels of BAX (r = 0.69, P < 0.05) and PTEN (r = 0.57, P < 0.05) gradually increased. It is concluded that TIEG1 can indue apoptosis of K562 cells and inhibit K562 cell proliferation in time-and dose-dependent manner. In apoptosis process of K562 cells induced by TIEG1, the expression changes of BCL-2/BAX and PTEN associate with the K562 cell apoptosis.


Subject(s)
Apoptosis , Cell Proliferation , Humans , K562 Cells , Kruppel-Like Transcription Factors , Metabolism , PTEN Phosphohydrolase , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , bcl-2-Associated X Protein , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-231599

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of tetrandrine (TET) on zinc finger protein 139 (ZNF139) and multidrug resistance (MDR) of human gastric carcinoma cell lines and possible mechanisms.</p><p><b>METHODS</b>Cultured SGC7901 and SGC7901/ADR were treated with TET (0.5, 1.0, 1.5, 2.0, and 2.5 microg/mL), then inhibition rates were measured by MTT assay in vitro. The expressions of ZNF139, MRP-1, MDR1, and GST-pi were detected by RT-PCR. The correlation between ZNF139 and each multidrug resistance factor was analyzed using Spearman correlation analysis, and the coefficient correlation was calculated.</p><p><b>RESULTS</b>The inhibition rate of TET (< or = 2.0 microg/mL) for SGC7901 and SGC7901/ADR was less than 10% with MTT assay. Expressions of ZNF139, MRP-1, MDR1, and GST-pi mRNA were higher in SGC7901/ADR than in SGC7901 (all P < 0.05). The expressions of ZNF139, MRP-1, MDR1, and GST--pi were down-regulated in SGC7901/ADR cells efficiently (all P < 0.01). Positive correlation existed between ZNF139 and MRP-1, ZNF139 and MDR1 before treated by TET in SGC7901/ADR, and this relationship also existed in SGC7901/ADR cells after treated by TET (all P < 0.05).</p><p><b>CONCLUSION</b>TET could achieve MDR reversion in gastric cancer cells by down-regulating the expression of ZNF139, MRP-1, and MDR1.</p>


Subject(s)
ATP Binding Cassette Transporter, Subfamily B , Metabolism , Benzylisoquinolines , Pharmacology , Cell Line, Tumor , Drug Resistance, Multiple , Genetics , Drug Resistance, Neoplasm , Genetics , Humans , Kruppel-Like Transcription Factors , Metabolism , Multidrug Resistance-Associated Proteins , Metabolism , Stomach Neoplasms , Metabolism , Zinc Fingers , Genetics
18.
Article in English | WPRIM | ID: wpr-251357

ABSTRACT

The clinical characteristics of patients with disorders of sex development (DSD), and the diagnostic values of classic cytogenetic and molecular genetic assays for DSD were investigated. In the enrolled 56 cases, there were 9 cases of 46,XY DSD, 6 cases of Turner syndrome (TS), one case of Super female syndrome, 25 cases of Klinefelter syndrome, 14 cases of 46,XX DSD, and one case of autosomal balanced rearrangements with hypospadias. The diagnosis of sex was made through physical examination, cytogenetic assay, ultrasonography, gonadal biopsy and hormonal analysis. PCR was used to detect SRY, ZFX, ZFY, DYZ3 and DYZ1 loci on Y and X chromosomes respectively. The DSD patients with the same category had similar clinical characteristics. The karyotypes in peripheral blood lymphocytes of all patients were identified. PCR-based analysis showed presence or absence of the X/Y-linked loci in several cases. Of the 9 cases of 46,XY DSD, 6 were positive for SRY, 9 for ZFX/ZFY, 9 for DYZ3 and 8 for DYZ1 loci. Of the 6 cases of TS, only 1 case with the karyotype of 45,X,/46,XX/46,XY was positive for all 5 loci. Of the 25 cases of Klinefelter syndrome, all were positive for all 5 loci. In one case of rare Klinefelter syndrome variants azoospermia factor (AZF) gene detection revealed the loss of the AZFa+AZFb region. In 14 cases of 46,XX DSD, 7 cases were positive for SRY, 14 for ZFX, 7 for ZFY, 7 for ZYZ3, and 5 for DYZ1. PCR can complement and also confirm cytogenetic studies in the diagnosis of sex in cases of DSD.


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Chromosome Aberrations , Chromosome Banding , Chromosomes, Human, X , Genetics , Chromosomes, Human, Y , Genetics , Disorders of Sex Development , Diagnosis , Genetics , Female , Gene Deletion , Genetic Loci , Genetics , Humans , Karyotyping , Kruppel-Like Transcription Factors , Genetics , Male , Polymerase Chain Reaction , Sex Chromosome Aberrations , Sex-Determining Region Y Protein , Genetics , Young Adult
19.
Article in Chinese | WPRIM | ID: wpr-254474

ABSTRACT

<p><b>OBJECTIVE</b>To study the frequency of rare blood group Lu(a-b-) phenotype in a population from Shanghai region, and to explore the molecular basis of Lu(a-b-) by detecting the Lu and Lu relative mediator gene EKLF/KLF1.</p><p><b>METHODS</b>Donors from Shanghai region were screened for Lutheran blood group by monoclonal anti-Lub using serological methods. Individuals with Lu(b-) were determined Lua, P1 and i antigens. Fifteen exons of the LU gene and 3 exons of the EKLF/KLF1 gene for the identified Lu(a-b-) samples were amplified and sequenced.</p><p><b>RESULTS</b>Ten Lu(a-b-) donors were obtained from 44 331 donors from Shanghai region. No homozygous or heterozygous mutations were found in the LU gene, whilst 7 mutations in EKLF/KLF1 gene were identified in the 10 samples.</p><p><b>CONCLUSION</b>The frequency of rare Lu(a-b-) blood group in Shanghai was approximately 0.02%, and all the individuals had an In(Lu) phenotype. The molecular basis of such samples may be related to mutations in the EKLF/KLF1 gene.</p>


Subject(s)
China , Ethnology , Humans , Kruppel-Like Transcription Factors , Genetics , Lutheran Blood-Group System , Genetics , Mutation , Phenotype
20.
Article in English | WPRIM | ID: wpr-175269

ABSTRACT

Prostate cancer is the most frequently diagnosed malignancy and the second leading cause of cancer mortality among men in the United States. Accumulating evidence suggests that lysophosphatidic acid (LPA) serves as an autocrine/paracrine mediator to affect initiation, progression and metastasis of prostate cancer. In the current study, we demonstrate that LPA stimulates migration and proliferation of highly metastatic human prostate cancer, PC-3M-luc-C6 cells. LPA-induced migration of PC-3M-luc-C6 cells was abrogated by pretreatment of PC-3M-luc-C6 cells with the LPA receptor 1/3 inhibitor Ki16425 or small interfering RNA (siRNA)-mediated silencing of endogenous LPA receptor 1, implicating a key role of the LPA-LPA receptor 1 signaling axis in migration of PC-3M-luc-C6 cells. In addition, LPA treatment resulted in augmented expression levels of Kruppel-like factor 4 (KLF4), and siRNA or short-hairpin RNA (shRNA)-mediated silencing of KLF4 expression resulted in the abolishment of LPA-stimulated migration and proliferation of PC-3M-luc-C6 cells. shRNA-mediated silencing of KLF4 expression resulted in the inhibition of in vivo growth of PC-3M-luc-C6 cells in a xenograft transplantation animal model. Taken together, these results suggest a key role of LPA-induced KLF4 expression in cell migration and proliferation of prostate cancer cells in vitro and in vivo.


Subject(s)
Animals , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Silencing , Humans , Kruppel-Like Transcription Factors/genetics , Lysophospholipids/metabolism , Male , Mice, Inbred BALB C , Prostate/metabolism , Prostatic Neoplasms/genetics
SELECTION OF CITATIONS
SEARCH DETAIL