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1.
Int J Mol Med ; 48(2)2021 Aug.
Article in English | MEDLINE | ID: mdl-34165154

ABSTRACT

Atherosclerosis (AS) is the main pathological basis of cardiovascular diseases, which are related to high morbidity and mortality rates. The present study aimed to investigate the role of the Krüppel­like factor 5 (KLF5)/LINC00346/miR­148a­3p loop in AS. The expression levels of KLF5 in serum and of KLF5/LINC00346/miR­148a­3p in human umbilical vein endothelial cells (HUVECs) were detected by RT­qPCR analysis. The protein expression levels of KLF5, phosphorylated (p­)endothelial nitric oxide synthase (eNOS) and eNOS in HUVECs were analyzed by western blot analysis. Changes in the levels of TNF­α, IL­1ß, IL­6 and nitric oxide (NO) were determined in the supernatant through the application of available commercial kits. The binding of KLF5 to the promoter region of LINC00346 was verified by chromatin immunoprecipitation (ChIP)­PCR assay. The combinatory interaction between KLF5 and LINC00346, LINC00346 and miR­148a­3p, and miR­148a­3p and KLF5 was confirmed by luciferase reporter assay. The results revealed that KLF5 expression was increased in the serum of patients with AS and also in oxidized low­density lipoprotein (OX­LDL)­stimulated HUVECs. The transcription factor KLF5 promoted the transcription of LINC00346. KLF5 interference or LINC00346 interference inhibited the expression of inflammatory factors and functional injury in OX­LDL­stimulated HUVECs. LINC00346 functioned as a sponge of miR­148a­3p. miR­148a­3p overexpression inhibited the expression of inflammatory factors and functional injury in OX­LDL­stimulated HUVECs and miR­148a­3p targeted KLF5 expression. On the whole, the present study demonstrates that KLF5 interference induces the downregulation of LINC00346 and also inhibits inflammation and functional injury in OX OX­LDL­stimulated HUVECs by upregulating miR­148a­3p expression.


Subject(s)
Atherosclerosis/genetics , Gene Expression Regulation , Human Umbilical Vein Endothelial Cells/metabolism , Inflammation/genetics , Kruppel-Like Transcription Factors/genetics , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Adult , Aged , Atherosclerosis/blood , Atherosclerosis/metabolism , Blotting, Western , Cells, Cultured , Cytokines/metabolism , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Inflammation/metabolism , Kruppel-Like Transcription Factors/blood , Kruppel-Like Transcription Factors/metabolism , Lipoproteins, LDL/pharmacology , Middle Aged , Nitric Oxide/metabolism , Reverse Transcriptase Polymerase Chain Reaction
2.
Sci Rep ; 11(1): 221, 2021 01 08.
Article in English | MEDLINE | ID: mdl-33420235

ABSTRACT

One epigenetic hallmark of many cancer types is differential DNA methylation occurring at multiple loci compared to normal tissue. Detection and assessment of the methylation state at a specific locus could be an effective cancer diagnostic. We assessed the effectiveness of hypermethylation at the CpG island of ZNF154, a previously reported multi-cancer specific signature for use in a blood-based cancer detection assay. To predict its effectiveness, we compared methylation levels of 3698 primary tumors encompassing 11 solid cancers, 724 controls, 2711 peripheral blood cell samples, and 350 noncancer disease tissues from publicly available methylation array datasets. We performed a single-molecule high-resolution DNA melt analysis on 71 plasma samples from cancer patients and 20 noncancer individuals to assess ZNF154 methylation as a candidate diagnostic metric in liquid biopsy and compared results to KRAS mutation frequency in the case of pancreatic carcinoma. We documented ZNF154 hypermethylation in early stage tumors, which did not increase in most noncancer disease or with respect to age or sex in peripheral blood cells, suggesting it is a promising target in liquid biopsy. ZNF154 cfDNA methylation discriminated cases from healthy donor plasma samples in minimal plasma volumes and outperformed KRAS mutation frequency in pancreatic cancer.


Subject(s)
DNA Methylation , Kruppel-Like Transcription Factors/blood , Kruppel-Like Transcription Factors/genetics , Neoplasms/genetics , Neoplasms/pathology , Adult , Biomarkers, Tumor/genetics , Case-Control Studies , Colonic Neoplasms/blood , Colonic Neoplasms/diagnosis , Colonic Neoplasms/genetics , Colonic Neoplasms/pathology , Epigenesis, Genetic , Female , Humans , Liquid Biopsy , Liver Neoplasms/blood , Liver Neoplasms/diagnosis , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Male , Middle Aged , Neoplasms/blood , Neoplasms/diagnosis , Ovarian Neoplasms/blood , Ovarian Neoplasms/diagnosis , Ovarian Neoplasms/pathology , Pancreatic Neoplasms/blood , Pancreatic Neoplasms/diagnosis , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/pathology
3.
Nat Commun ; 12(1): 24, 2021 01 05.
Article in English | MEDLINE | ID: mdl-33402679

ABSTRACT

Differences between sexes contribute to variation in the levels of fasting glucose and insulin. Epidemiological studies established a higher prevalence of impaired fasting glucose in men and impaired glucose tolerance in women, however, the genetic component underlying this phenomenon is not established. We assess sex-dimorphic (73,089/50,404 women and 67,506/47,806 men) and sex-combined (151,188/105,056 individuals) fasting glucose/fasting insulin genetic effects via genome-wide association study meta-analyses in individuals of European descent without diabetes. Here we report sex dimorphism in allelic effects on fasting insulin at IRS1 and ZNF12 loci, the latter showing higher RNA expression in whole blood in women compared to men. We also observe sex-homogeneous effects on fasting glucose at seven novel loci. Fasting insulin in women shows stronger genetic correlations than in men with waist-to-hip ratio and anorexia nervosa. Furthermore, waist-to-hip ratio is causally related to insulin resistance in women, but not in men. These results position dissection of metabolic and glycemic health sex dimorphism as a steppingstone for understanding differences in genetic effects between women and men in related phenotypes.


Subject(s)
Anorexia Nervosa/genetics , Blood Glucose/metabolism , Glucose Intolerance/genetics , Insulin Receptor Substrate Proteins/genetics , Insulin Resistance/genetics , Insulin/blood , Kruppel-Like Transcription Factors/genetics , Adult , Anorexia Nervosa/blood , Anorexia Nervosa/ethnology , Anorexia Nervosa/physiopathology , Fasting/blood , Female , Gene Expression , Genetic Loci , Genome-Wide Association Study , Glucose Intolerance/blood , Glucose Intolerance/ethnology , Glucose Intolerance/physiopathology , Humans , Insulin Receptor Substrate Proteins/blood , Kruppel-Like Transcription Factors/blood , Male , Middle Aged , Phenotype , Sex Characteristics , Sex Factors , Waist-Hip Ratio , White People
4.
J Exp Clin Cancer Res ; 39(1): 193, 2020 Sep 21.
Article in English | MEDLINE | ID: mdl-32958011

ABSTRACT

BACKGROUND: Angiogenesis, a basic requirement for tumor cell survival, is considered to be a malignant characteristic of small cell lung cancer (SCLC) and is closely related to the poor outcomes of SCLC patients. miR-141 has been found to play pro- and antiangiogenic roles in different cancers, but its role in SCLC angiogenesis has never been explored. METHODS: Total RNA was isolated from plasm exosomes and serum of SCLC patients to examine the expression of miR-141 by qRT-PCR. Cell proliferation, invasion, migration, tube formation assay, aortic ring assay and mouse tumor model were used to investigate the effect of exosomal miR-141 in angiogenesis in vitro and in vivo. Dual-luciferase assay was conducted to explore the target gene of miR-141. RESULTS: Circulating miR-141 was upregulated in samples from 122 SCLC patients compared with those from normal volunteers and that the increase in miR-141 was significantly associated with advanced TNM stages, implying the potential oncogenic role of miR-141 in SCLC malignancy. In vitro, miR-141 that was packaged into SCLC cell-secreted exosomes and delivered to human umbilical vein vascular endothelial cells (HUVECs) via exosomes facilitated HUVEC proliferation, invasion, migration and tube formation and promoted microvessel sprouting from mouse aortic rings. Matrigel plug assays demonstrated that SCLC cell-derived exosomal miR-141 induced neoangiogenesis in vivo. Furthermore, mouse subcutaneous tumor nodules that were developed from miR-141-overexpressing SCLC cells had a higher microvessel density (MVD) and grew faster than those developed from negative control cells. KLF12 was found to be the direct target gene of miR-141 and that the proangiogenic effect of miR-141 on HUVECs was abrogated by KLF12 overexpression. CONCLUSIONS: Our results demonstrate the specific function of the exosomal miR-141/KLF12 pathway in SCLC angiogenesis for the first time and provide potential novel targets for antiangiogenic therapies for SCLC patients.


Subject(s)
Kruppel-Like Transcription Factors/genetics , MicroRNAs/genetics , Neovascularization, Pathologic/genetics , Small Cell Lung Carcinoma/genetics , Animals , Cell Movement/genetics , Cell Proliferation/genetics , Exosomes/genetics , Female , Gene Expression Regulation, Neoplastic/genetics , Heterografts , Human Umbilical Vein Endothelial Cells , Humans , Kruppel-Like Transcription Factors/blood , Male , Mice , MicroRNAs/blood , Microvessels/growth & development , Microvessels/pathology , Middle Aged , Neovascularization, Pathologic/blood , Neovascularization, Pathologic/pathology , Signal Transduction/genetics , Small Cell Lung Carcinoma/blood , Small Cell Lung Carcinoma/pathology
5.
Cell Death Dis ; 11(9): 788, 2020 09 23.
Article in English | MEDLINE | ID: mdl-32968054

ABSTRACT

Circular RNAs (circRNAs) are a newly discovered type of biological molecule that belongs to the noncoding RNA family. Abundant evidence has shown that circRNAs are involved in the progression of various cancers. However, the particular functions of circRNAs in colorectal cancer (CRC) remain elusive. In this study, we investigated the differentially expressed circRNAs in three pairs of cancer tissue and adjacent normal tissue of CRC. We revealed that circGLIS2 expression was higher in CRC tissue and cell lines. Gain-and-loss function assays showed that circGLIS2 was involved in the regulation of cell migration. Moreover, overexpressing circGLIS2 in CRC cells activated the NF-κB pathway and induced pro-inflammatory chemokine production, which evoked tumor-associated inflammation through recruiting leukocytes. In turn, when the cancer cells were exposed to the supernatant of circGLIS2 overexpressed cancer cells, they were endowed with the ability of migration and chemokines production. Furthermore, the rescue assay confirmed that circGLIS2 activated NF-κB signaling and promoted cell migration by sponging miR-671. Overall, our study reveals that circGLIS2, acting as a potential oncogene, maintains the abnormal activation state of the NF-κB signaling pathway via the miR-671 sponge mechanism in CRC cells. This study provides a scientific basis for targeting circGLIS2 in colorectal cancer interventions.


Subject(s)
Colonic Neoplasms/genetics , Colorectal Neoplasms/genetics , Kruppel-Like Transcription Factors/blood , NF-kappa B/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Colonic Neoplasms/blood , Colorectal Neoplasms/blood , Disease Progression , Gene Expression Regulation, Neoplastic/genetics , Humans , Kruppel-Like Transcription Factors/genetics , RNA, Circular
6.
Mol Med Rep ; 22(2): 1429-1439, 2020 08.
Article in English | MEDLINE | ID: mdl-32626976

ABSTRACT

Zinc finger protein 804A (ZNF804A) has been identified by genome­wide association studies as a robust risk gene in schizophrenia, but how ZNF804A contributes to schizophrenia and its upstream regulation remains unknown. Previous studies have indicated that microRNAs (miRs) are key factors that regulate the expression levels of their target genes. The present study revealed significantly increased expression of miR­148b­3p in the peripheral blood of patients with first­onset schizophrenia compared with healthy controls, and bioinformatics analysis predicted that the ZNF804A gene is a target of miR­148b­3p. Therefore, the present study investigated the possible upstream regulation of ZNF804A by miR­148b­3p in the human neuroblastoma SH­SY5Y cell line, and assessed the implications for schizophrenia. The results revealed significantly reversed expression levels of miR­148b­3p (P=0.0051) and ZNF804A (P=0.0218) in the peripheral blood of patients with first­onset schizophrenia compared with healthy individuals. Furthermore, it was demonstrated that miR­148b­3p directly targeted ZNF804A via binding to conserved target sites in the 3'­untranslated region of ZNF804A mRNA, where it inhibited the endogenous expression of ZNF804A at both the mRNA (P=0.048) and protein levels (P=0.013) in SH­SY5Y cells. Furthermore, miR­148b­3p was revealed to regulate the expression levels of catechol­O­methyltransferase (COMT) and serine protease 16 (PRSS16) by targeting ZNF804A in SH­SY5Y cells. Collectively, the present results indicated that there was a direct upstream regulation of the schizophrenia risk gene ZNF804A by miR­148b­3p, which contributed to the regulation of the downstream genes COMT and PRSS16. Thus, the miR­148b­3p/ZNF804A/COMT/PRSS16 pathway may play an important role in the pathophysiology of schizophrenia, and may serve as a potential target in drug discovery and gene therapy for this disorder.


Subject(s)
Catechol O-Methyltransferase/metabolism , Kruppel-Like Transcription Factors/metabolism , MicroRNAs/metabolism , Schizophrenia/metabolism , Serine Endopeptidases/metabolism , Case-Control Studies , Cell Line, Tumor , Gene Expression Regulation , Humans , Kruppel-Like Transcription Factors/blood , MicroRNAs/blood
7.
Ann Vasc Surg ; 68: 338-343, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32360698

ABSTRACT

BACKGROUND: Krüppel-like factor 4 (KLF4) is known to preserve vascular homeostasis. In the present study, we sought to correlate serum KLF4 levels with arterial aneurysm size and their clinical presentation. We also explored the association between serum KLF4 levels and the severity of extracranial carotid and peripheral arterial disease. METHODS: Patients undergoing surgery for various forms of atheromatosis (ATH group) or for arterial aneurysm repair (AA group) were eligible for inclusion. KLF4 levels were measured via enzyme-linked immunosorbent assay. RESULTS: Patients in the atheromatic and aneurysmal groups had significantly higher serum KLF4 levels compared with controls. Patients with permanent end-organ damage (ATH3) had higher serum KLF4 (6.96 ± 0.75 pg/mL) compared with patients with asymptomatic internal carotid stenosis >70% or claudication (ATH1) (2.76 ± 0.68 pg/mL; mean difference [MD], -4.20; 95% confidence interval [95% CI], -5.35 to -3.04; P < 0.01) and those with transient ischemic attack or rest pain (ATH2) (4.47 ± 1.08 pg/mL; MD, -2.48; 95% CI, -3.76 to -1.21). Furthermore, patients with an asymptomatic aneurysm of a diameter 250-300% of that of the normal artery (AA1, 5.01 ± 1.08 pg/mL) had considerably lower serum KLF4 compared with those suffering from either a symptomatic aneurysm or an asymptomatic aneurysm of a diameter >350% of that of normal artery (AA3, 6.63 ± 1.92 pg/mL; MD, -2.61; 95% CI, -5.04 to -0.18; P < 0.01). CONCLUSIONS: Serum KLF4 levels are significantly increased in patients with end-organ damage related to atheromatosis as well as those with extensive aneurysmal disease.


Subject(s)
Aneurysm/blood , Carotid Stenosis/blood , Kruppel-Like Transcription Factors/blood , Peripheral Arterial Disease/blood , Aneurysm/diagnostic imaging , Carotid Stenosis/diagnostic imaging , Humans , Kruppel-Like Factor 4 , Peripheral Arterial Disease/diagnostic imaging , Retrospective Studies , Severity of Illness Index , Up-Regulation
8.
Forensic Sci Int ; 311: 110267, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32325350

ABSTRACT

Many studies in the forensic field have reported that analysis of DNA methylation is the most reliable method of predicting age. In a previous study, 5 CpG sites located in ELOVL2, FHL2, KLF14, C1orf132 and TRIM59 genes were tested for age prediction purposes in blood, saliva and buccal swab samples from Korean individuals using a multiplex methylation SNaPshot assay. The main goals of the present study were i) to replicate the same multiplex SNaPshot assay in blood samples from Portuguese individuals, ii) to compare DNA methylation status between two different populations and iii) to address putative differences in the methylation status between blood from living and deceased individuals. Blood samples from 59 living individuals (37 females, 22 males; aged 1-94 years-old) and from 62 deceased individuals (13 females, 49 males; aged 28-86 years-old) were evaluated. The specific primers were those previously described. Linear regression models were used to analyse relationships between methylation levels and chronological age using IBM SPSS software v.24. Our results allowed to build a final age prediction model (APM) for blood samples of living individuals with 3 CpG sites, at ELOVL2, FHL2 and C1orf132 genes, explaining 96.3% of age variation, with a mean absolute deviation (MAD) from chronological age of 4.25 years. Some differences were found in the extent of the age association in the targeted loci comparing Portuguese with Korean individuals. The final APM built for deceased individuals included 4 CpG sites, at ELOVL2, FHL2, C1orf132 and TRIM59 genes, explaining 79.3% of age variation, with a MAD of 5.36 years. Combining both sets of samples from living and deceased individuals, the most accurate APM with 4 CpGs, at ELOVL2, FHL2, C1orf132 and TRIM59 genes, explained 92.5% of variation in age, with a MAD of 4.97 years. In conclusion, our study replicated in blood samples of Portuguese living individuals a previous SNaPshot assay for age estimation. The possibility that age markers might be population specific and that postmortem changes can alter the methylation status among specific loci was suggested by our data. Our study showed the usefulness of the multiplex methylation SNaPshot assay for forensic analysis in blood samples of living and deceased individuals.


Subject(s)
Aging/genetics , DNA Methylation , Forensic Genetics/methods , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , CpG Islands , Fatty Acid Elongases/blood , Female , Genetic Markers , Genotyping Techniques/instrumentation , Humans , Infant , Intracellular Signaling Peptides and Proteins/blood , Kruppel-Like Transcription Factors/blood , LIM-Homeodomain Proteins/blood , Linear Models , Male , Middle Aged , Muscle Proteins/blood , Portugal , Transcription Factors/blood , Tripartite Motif Proteins/blood , Young Adult
9.
Int J Cancer ; 147(6): 1740-1752, 2020 09 15.
Article in English | MEDLINE | ID: mdl-32191343

ABSTRACT

Accumulated evidence revealed that aberrant CpG island hypermethylation plays an important role in carcinogenesis which can serve as a promising target for molecular detection in body fluids. Despite a myriad of attempts to diagnose ovarian cancer (OC) at an early stage, this clinical aim remains a major challenge. To date, no single biomarker is able to accurately detect early OC in either tissue or body fluid. Aberrant DNA methylation patterns in circulating DNA provide highly specific cancer signals. In our study, we establish a novel panel of methylation-specific genes for the development of a TaqMan based qPCR assay to quantify methylation levels. We analyzed promoter methylation of homeobox A9 (HOXA9) and hypermethylated in cancer 1 (HIC1) quantitatively in 120 tissue samples and in 70 matched serum cell-free DNA (CFDNA) of cancerous and noncancerous samples by MethyLight assay. HOXA9 and HIC1 methylation occurred in 82.3 and 80.0% of OC tissue samples in singleplex assay, thereby confirming that methylation was highly cancer-specific. When either or both gene promoter showed methylation, the sensitivity was 88.2% with a specificity of 88.6% in tissue samples. The combined sensitivity for this novel marker panel in serum CFDNA was 88.9% (area under the curve [AUC] = 0.95). In contrast, no hypermethylation was observed in serum from matched cancer-free control women. Our results confirm the elevated performance of novel epigenetic marker panel (HOXA9 and HIC1) when analyzed in tissue and matched serum samples. Our findings reveal the potential of this biomarker panel as a suitable diagnostic serum biomarker for early screening of OC.


Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Ovarian Epithelial/diagnosis , Early Detection of Cancer/methods , Homeodomain Proteins/genetics , Kruppel-Like Transcription Factors/genetics , Ovarian Neoplasms/diagnosis , Adolescent , Adult , Aged , Biomarkers, Tumor/blood , Carcinoma, Ovarian Epithelial/blood , Carcinoma, Ovarian Epithelial/genetics , Carcinoma, Ovarian Epithelial/surgery , Circulating Tumor DNA/blood , Circulating Tumor DNA/genetics , CpG Islands/genetics , DNA Methylation , Early Detection of Cancer/instrumentation , Epigenesis, Genetic , Epigenomics/instrumentation , Epigenomics/methods , Feasibility Studies , Female , Gene Expression Regulation, Neoplastic , Homeodomain Proteins/blood , Humans , Kruppel-Like Transcription Factors/blood , Liquid Biopsy/methods , Middle Aged , Ovarian Neoplasms/blood , Ovarian Neoplasms/genetics , Ovary/pathology , Ovary/surgery , Predictive Value of Tests , Preoperative Period , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction/instrumentation , Young Adult
10.
Cancer Med ; 9(8): 2803-2811, 2020 04.
Article in English | MEDLINE | ID: mdl-32101374

ABSTRACT

BACKGROUND: Universal gene targets are in persistent demand by real-time quantitative polymerase chain reaction (RT-qPCR)-based methods in acute leukemia (AL) diagnosis and monitoring. Human Krüppel-like factor 3 (hKLF3), a newly cloned human transcription factor, has proved to be a regulator of hematopoiesis. METHODS: Sanger sequencing was performed in bone marrow (BM) samples from 17 AL patients for mutations in hKLF3 coding exons. hKLF3 expression in peripheral blood (PB) and BM samples from 45 AL patients was dynamically detected by RT-qPCR. PB samples from 31 healthy donors were tested as normal controls. RESULTS: No mutation was sequenced in hKLF3 coding exons. hKLF3 expression in PB of AL was significantly lower than that in healthy donors [0.30 (0.02-1.07) vs 1.18 (0.62-3.37), P < .0001]. Primary acute myeloid leukemia (AML) exhibited the least expression values compared with secondary AML and acute lymphoblastic leukemia. Receiver operating characteristic (ROC) analyses suggested that hKLF3 expression in PB was a good marker for AML diagnosis with an AUC of 0.99 (95% CI 0.98-1.00) and an optimum cutoff value of 0.67 (sensitivity 93.94% and specificity 93.55%). hKLF3 expression was upregulated significantly when AML patients acquired morphological complete remission (CR), and the level of hKLF3 seemed to be higher in patients with deeper CR than in patients with minimal residual disease (MRD). Paired PB and BM samples showed highly consistent alteration in hKLF3 expression (r = .6533, P = .001). Besides, a significantly converse correlation between decreased hKLF3 expression in PB and markers for leukemic load was observed. CONCLUSIONS: hKLF3 expression in PB may act as a potential marker for AL diagnosis and monitoring.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/blood , Kruppel-Like Transcription Factors/blood , Leukemia, Myeloid, Acute/pathology , Neoplasm, Residual/pathology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Adolescent , Adult , Case-Control Studies , Female , Follow-Up Studies , Humans , Leukemia, Myeloid, Acute/blood , Leukemia, Myeloid, Acute/drug therapy , Male , Middle Aged , Neoplasm, Residual/blood , Neoplasm, Residual/drug therapy , Precursor Cell Lymphoblastic Leukemia-Lymphoma/blood , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Prognosis , Young Adult
11.
Clin Chim Acta ; 495: 630-636, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31153870

ABSTRACT

BACKGROUND: Current assessment tools for patients with acute chest pain are either traumatic (coronary angiography) or unreliable (measurement of cardiac troponin concentrations). We investigated whether the novel cardiovascular stress markers, serum growth differentiation factor-15 (GDF-15), Krüppel-like factor 4 (KLF4) and growth arrest-specific 6 (gas6) may be useful biomarkers of coronary artery disease (CAD). METHODS: A total of 350 male patients were enrolled, 198 with CAD and 152 controls, based on coronary angiography. GDF-15, KLF4 and gas6 concentrations were measured using commercial enzyme-linked immunosorbent assay kits. Multivariate logistic regression and multivariate linear regression were performed to evaluate potential associations of GDF-15, KLF4 and gas6 with risk of CAD or CAD severity. RESULTS: Serum GDF-15, KLF4 and gas6 concentrations were significantly higher in male patients with CAD than in control subjects (P < .05), and they correlated significantly with involvement of coronary vessels (P < .05). After adjusting for confounding factors, we found that circulating GDF-15 concentrations remained positively associated with the presence of CAD (odds ratio [OR] per 1-standard deviation [SD] increase, 3.182; 95% confidence interval [CI] 1.586 to 6.382; P = .001), as did KLF4 concentrations (OR per 1-SD increase, 13.05; 95% CI 2.940 to 57.921, P = .001). Moreover, circulating GDF-15 concentrations were positively associated with the Gensini score (estimated SD change per 1-SD increase, 22.091; 95% CI 9.147 to 35.035, P = .001), as were KLF4 concentrations (estimated SD change per 1-SD increase, 27.996; 95% CI 10.082 to 45.910, P = .002). Gas6, in contrast, showed no relationship to presence of CAD or Gensini score. , CONCLUSIONS: In this case-control study, increased concentrations of circulating GDF-15 and KLF4 were significantly associated with the presence and severity of CAD.


Subject(s)
Coronary Artery Disease/blood , Growth Differentiation Factor 15/blood , Intercellular Signaling Peptides and Proteins/blood , Kruppel-Like Transcription Factors/blood , Aged , Angiography , Biomarkers/blood , Coronary Artery Disease/diagnostic imaging , Early Diagnosis , Humans , Kruppel-Like Factor 4 , Male , Middle Aged
12.
J BUON ; 24(2): 599-607, 2019.
Article in English | MEDLINE | ID: mdl-31128012

ABSTRACT

PURPOSE: Non-small cell lung cancer (NSCLC) is still the commonest fatal malignancy worldwide. The relationship between miR-660-5p and progress of NSCLC has not been well confirmed in recent studies. This manuscript focused to the function of miR-660-5p during the appearance and progression of NSCLC. METHODS: To identify the expression level of miR-660-5p in NSCLC, patient plasma and exosomes, quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed. Cell proliferation and colony formation abilities were examined by Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Then, the influence of miR-660-5p on migration and invasion was analyzed by transwell assay. Bioinformatics and Luciferase report assay were used to find potential target genes. Western blot was chosen to assess the expression level of KLF9. Stably transfected NSCLC cells (A549 and H1299) were injected into nude mice to identify the function of miR-660-5p in tumorigenesis in vivo. RESULTS: Compared with healthy controls, the release of miR-660-5p in plasma and exosomes was increased in patients with NSCLC (n=80). Knockdown of miR-660-5p significantly suppressed proliferation, migration, and invasion, whereas overexpression of miR-660-5p had the opposite effect. KLF9 might be a potential target of miR-660-5p. In addition, up-regulation of miR-660-5p promoted tumorigenesis in vivo, and the protein level of KLF9 also decreased in xenografts. CONCLUSIONS: Our current study suggests that miR-660-5p may control NSCLC proliferation, viability, and metastasis by targeting KLF9, which provides a potential therapeutic target for NSCLC.


Subject(s)
Carcinogenesis/genetics , Carcinoma, Non-Small-Cell Lung/genetics , Kruppel-Like Transcription Factors/genetics , MicroRNAs/genetics , A549 Cells/metabolism , Animals , Carcinoma, Non-Small-Cell Lung/blood , Carcinoma, Non-Small-Cell Lung/pathology , Cell Movement/genetics , Cell Proliferation/genetics , Exosomes/genetics , Gene Expression Regulation, Neoplastic , Heterografts , Humans , Kruppel-Like Transcription Factors/blood , Mice , MicroRNAs/blood , Neoplasm Metastasis
13.
Clin Lab ; 65(4)2019 Apr 01.
Article in English | MEDLINE | ID: mdl-30969076

ABSTRACT

BACKGROUND: Kruppel-like factor 7 (KLF7) is associated with type 2 diabetes and obesity; however, at present the KLF7 level in polycystic ovary syndrome (PCOS) remains unreported. METHODS: In this study, serum KLF7, glucose, insulin, C-reactive protein (CRP), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and total testosterone concentrations were measured in 65 women with PCOS and 61 healthy controls. RESULTS: Body mass index (BMI) in PCOS and the control group were all < 25 kg/m2. The median concentration of KLF7 was 3.630 ng/mL [interquartile range (IQR): 1.547 - 7.172] in women with PCOS, which was significantly lower than that of controls (5.282 ng/mL, IQR: 3.128 - 11.263, p = 0.003). The KLF7 level was positively correlated with low-density lipoprotein cholesterol (LDL-C) (r = 0.261, p = 0.018). No significant association was observed between KLF7 and the BMI, total testosterone, and insulin resistance (IR). CONCLUSIONS: The KLF7 level decreased in women with PCOS. KLF7 may represent a new therapeutic target in the treatment of PCOS.


Subject(s)
Cholesterol, LDL/blood , Kruppel-Like Transcription Factors/blood , Polycystic Ovary Syndrome/blood , Adult , Blood Glucose/metabolism , Case-Control Studies , Female , Humans , Inflammation , Insulin/blood , Insulin Resistance , Luteinizing Hormone/blood , Obesity/blood , Testosterone/blood , Young Adult
14.
Biochem Pharmacol ; 164: 115-128, 2019 06.
Article in English | MEDLINE | ID: mdl-30954486

ABSTRACT

Uric acid (UA) is a promising protective treatment in ischaemic stroke, but the precise molecular targets underlying its in vivo beneficial actions remain unclear. High concentrations of UA inhibit angiogenesis of cultured endothelial cells via Krüppel-like factor 2 (KLF)-induced downregulation of vascular endothelial growth factor (VEGF), a pro-angiogenic mediator that is able to increase blood-brain barrier (BBB) permeability in acute stroke. Here, we investigated whether UA treatment after ischaemic stroke protects brain endothelial cell functions and modulates the KLF2-VEGF-A axis. Transient intraluminal middle cerebral artery (MCA) occlusion/reperfusion was induced in adult male spontaneously hypertensive (SHR) rats and corresponding normotensive Wistar-Kyoto (WKY) rats. Animals received UA (16 mg/kg) or vehicle (Locke's buffer) i.v. at reperfusion. BBB permeability was evaluated by Evans blue extravasation to the brain and in human cerebral endothelial hCMEC/D3 cells under oxygen-glucose deprivation/re-oxygenation. Circulating VEGF-A levels were measured in rats and acute ischaemic stroke patients from the URICO-ICTUS trial. Angiogenesis progression was assessed in Matrigel-cultured MCA. Worse post-stroke brain damage in SHR than WKY rats was associated with higher hyperaemia at reperfusion, increased Evans blue extravasation, exacerbated MCA angiogenic sprouting, and higher VEGF-A levels. UA treatment reduced infarct volume and Evans blue leakage in both rat strains, improved endothelial cell barrier integrity and KLF2 expression, and lowered VEGF-A levels in SHR rats. Hypertensive stroke patients treated with UA showed lower levels of VEGF-A than patients receiving vehicle. Consistently, UA prevented the enhanced MCA angiogenesis in SHR rats by a mechanism involving KLF2 activation. We conclude that UA treatment after ischaemic stroke upregulates KLF2, reduces VEGF-A signalling, and attenuates brain endothelial cell dysfunctions leading to neuroprotection.


Subject(s)
Blood-Brain Barrier/metabolism , Hypertension/blood , Kruppel-Like Transcription Factors/blood , Stroke/blood , Uric Acid/therapeutic use , Vascular Endothelial Growth Factor A/blood , Animals , Antioxidants/pharmacology , Antioxidants/therapeutic use , Biomarkers/blood , Blood-Brain Barrier/drug effects , Brain/drug effects , Brain/metabolism , Cell Line , Double-Blind Method , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Humans , Hypertension/drug therapy , Hypertension/pathology , Kruppel-Like Transcription Factors/agonists , Male , Neuroprotective Agents/pharmacology , Neuroprotective Agents/therapeutic use , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Stroke/drug therapy , Stroke/pathology , Treatment Outcome , Uric Acid/pharmacology , Vascular Endothelial Growth Factor A/antagonists & inhibitors
15.
J Mol Cell Cardiol ; 114: 29-37, 2018 01.
Article in English | MEDLINE | ID: mdl-29061339

ABSTRACT

Physiological changes during embryonic development are associated with changes in the isoform expression of both myocyte sarcomeric proteins and of erythrocyte haemoglobins. Cell type-specific isoform expression of these genes also occurs. Although these changes appear to be coordinated, it is unclear how changes in these disparate cell types may be linked. The transcription factor Hic2 is required for normal cardiac development and the mutant is embryonic lethal. Hic2 embryos exhibit precocious expression of the definitive-lineage haemoglobin Hbb-bt in circulating primitive erythrocytes and of foetal isoforms of cardiomyocyte genes (creatine kinase, Ckm, and eukaryotic elongation factor Eef1a2) as well as ectopic cardiac expression of fast-twitch skeletal muscle troponin isoforms. We propose that HIC2 regulates a switching event within both the contractile machinery of cardiomyocytes and the oxygen carrying systems during the developmental period where demands on cardiac loading change rapidly.


Subject(s)
Cardiovascular System/embryology , Cardiovascular System/metabolism , Kruppel-Like Transcription Factors/metabolism , Protein Isoforms/metabolism , Tumor Suppressor Proteins/metabolism , Animals , Embryo Loss/pathology , Erythrocytes/metabolism , Fetus/metabolism , Gene Expression Regulation, Developmental , Hemoglobins/metabolism , Kruppel-Like Transcription Factors/blood , Mice , Mutation/genetics , Myocytes, Cardiac/metabolism , Organ Specificity , Time Factors , Troponin I/metabolism , Tumor Suppressor Proteins/blood
16.
World Neurosurg ; 108: 909-916.e3, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28893694

ABSTRACT

BACKGROUND: Cerebral vasospasm (CVS) is a major determinant of prognosis in patients with subarachnoid hemorrhage (SAH). Alteration in the vascular phenotype contributes to development of CVS. However, little is known about the role of microRNAs (miRNAs) in the phenotypic alteration after SAH. We investigated the expression profile of miRNAs and the chronologic changes in the expression of microRNA-15a (miR-15a) and Kruppel-like factor 4 (KLF4), a potent regulator of vascular phenotype modulation that modulates the expression of miR-15a, in the plasma and cerebrospinal fluid (CSF) of patients with SAH. METHODS: Peripheral blood and CSF samples were collected from 8 patients with aneurysmal SAH treated with endovascular obliteration. Samples obtained from 3 patients without SAH were used as controls in the analysis. Exosomal miRNAs were isolated and subjected to microarray analysis with the three-dimensional-gene miRNA microarray kit. The time course of the expression of miR-15a and KLF4 was analyzed using quantitative real-time polymerase chain reaction. RESULTS: Microarray analysis showed that 12 miRNAs including miR-15a were upregulated or downregulated both in the CSF and in plasma after SAH within 3 days. Quantitative real-time polymerase chain reaction showed that miR-15a expression was significantly increased in both the CSF and plasma, with a peak around 3-5 days after SAH, whereas the expression of KLF4 was significantly decreased around 1-3 days after SAH and remained lower than in controls. CONCLUSIONS: Our results suggest that an early and persistent decrease in KLF4 followed by an increase in miR-15a may contribute to the altered vascular phenotype, resulting in development of CVS.


Subject(s)
Kruppel-Like Transcription Factors/genetics , MicroRNAs/genetics , RNA, Messenger/cerebrospinal fluid , Subarachnoid Hemorrhage/genetics , Adult , Aged , Aged, 80 and over , Case-Control Studies , Down-Regulation , Female , Humans , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/blood , Kruppel-Like Transcription Factors/cerebrospinal fluid , Male , MicroRNAs/blood , MicroRNAs/cerebrospinal fluid , Microarray Analysis , Middle Aged , RNA, Messenger/blood , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Subarachnoid Hemorrhage/blood , Subarachnoid Hemorrhage/cerebrospinal fluid , Subarachnoid Hemorrhage/surgery , Up-Regulation
17.
Tumour Biol ; 39(5): 1010428317705339, 2017 May.
Article in English | MEDLINE | ID: mdl-28475008

ABSTRACT

Malignant glioma is one of the most common primary brain tumors that develop via multiple pathways and gene deregulation. MicroRNAs are involved in human cancer development and progression, and their serum expression profiles of glioma patients may be useful for classifying cancers. However, the profile and molecular mechanism of serum microRNAs for human glioma are poorly understood. Thus, it is crucial to analyze microRNA expression in human glioma serum to identify molecular subclasses and early stage of glioma. In this study, we performed microRNA alteration that contributes to glioma profile via analysis of The Cancer Genome Atlas RNA sequencing data and other independent Gene Expression Omnibus microarray data. We identified the glioma-associated novel microRNA as a key regulator of human glioma development and progression. The putative novel miR-1825 was validated by real-time polymerase chain reaction and its expression was significantly decreased in the serum of glioma patients compared with healthy controls. Patients with high miR-1825 expression had a longer survival rate. Interestingly, we found that miR-1825 expression levels were dependent on tumor size and pathological grading in glioma patients, but not associated with other factors including age and T classification. MicroRNA-Gene Ontology network indicated that miR-1825 may play an important role in the development of human glioma including apoptosis, cell proliferation, and invasion. In vitro assays of miR-1825 inhibit U87 cell proliferation and invasion and induce apoptosis. Furthermore, we provide evidence that the tumor-suppressive microRNA miR-1825 controls KLF2 expression. Reporter gene analyses revealed that both microRNAs directly targeted the 3'-untranslated region of KLF2 messenger RNA. These data demonstrated that miR-1825 expression in serum of human glioma was associated with tumorigenesis and miR-1825 may be used as a biomarker for identification of the pathological grade of glioma.


Subject(s)
Biomarkers, Tumor/blood , Glioma/blood , Kruppel-Like Transcription Factors/blood , MicroRNAs/blood , Adult , Apoptosis/genetics , Biomarkers, Tumor/genetics , Carcinogenesis/genetics , Cell Proliferation/genetics , Female , Gene Expression Regulation, Neoplastic , Glioma/genetics , Glioma/pathology , Humans , Kruppel-Like Transcription Factors/genetics , Male , MicroRNAs/genetics , Middle Aged , Neoplasm Staging
18.
J Clin Lipidol ; 11(1): 61-69, 2017.
Article in English | MEDLINE | ID: mdl-28391912

ABSTRACT

BACKGROUND: Persistent immune activation and inflammation in virologically suppressed human immunodeficiency virus (HIV) infection are linked to excess cardiovascular risk. OBJECTIVE: To evaluate atorvastatin as a strategy to reduce cardiovascular risk. METHODS: A5275 was a multicenter, prospective, randomized, double-blind, placebo-controlled, cross-over pilot study of atorvastatin (10 mg/day for 4 weeks then 20 mg/day for 16 weeks) with a planned enrollment of 97 HIV-infected participants ≥18 years old, receiving boosted protease inhibitor-based antiretroviral therapy for ≥6 months, with plasma HIV-1 RNAs below limits of quantification ≥180 days, and fasting low-density lipoprotein (LDL) cholesterol ≥70 and <130 mg/dL. Primary endpoints were differences of changes ([week 44-week 24]-[week 20-baseline]) in CD4+ and CD8+ T-lymphocyte activation (% CD38+/DR+) and plasma levels of IL-6 and D-dimer. Arms were compared using the Wilcoxon rank-sum tests and also summarized changes pre-to-post atorvastatin treatment. Analyses were as-treated. RESULTS: Ninety-eight participants were enrolled at 31 U S sites and 73 completed study treatment. Atorvastatin treatment did not decrease T-lymphocyte or monocyte activation, circulating biomarker levels (interleukin-6, D-dimer, soluble CD14, soluble CD163, monocyte chemoattractant protein-1, interferon-gamma-induced protein-10, high-sensitivity C-reactive protein, CD40L, and P-selectin) or white blood cell Krüppel-like Factor 2/4 messenger RNA levels. Pre-to-post atorvastatin reductions in calculated LDL (-38%), oxidized-LDL (-33%), and lipoprotein-associated phospholipase A2 (-31%) were significant (P < .01). CONCLUSION: In virologically suppressed individuals with HIV infection, atorvastatin did not significantly decrease levels of soluble or cellular biomarkers of immune activation and inflammation but resulted in robust reductions in LDL cholesterol, oxLDL, and lipoprotein-associated phospholipase A2, biomarkers associated with cardiovascular risk.


Subject(s)
Atorvastatin/pharmacology , Cholesterol, LDL/blood , HIV Infections/drug therapy , HIV Infections/immunology , HIV-1/physiology , Lipids/blood , Adult , Atorvastatin/adverse effects , Atorvastatin/therapeutic use , Biomarkers/blood , Female , HIV Infections/blood , Humans , Inflammation/blood , Kruppel-Like Transcription Factors/blood , Male , Middle Aged , Safety
19.
Development ; 144(3): 430-440, 2017 02 01.
Article in English | MEDLINE | ID: mdl-28143845

ABSTRACT

Transcription factor control of cell-specific downstream targets can be significantly altered when the controlling factor is mutated. We show that the semi-dominant neonatal anemia (Nan) mutation in the EKLF/KLF1 transcription factor leads to ectopic expression of proteins that are not normally expressed in the red blood cell, leading to systemic effects that exacerbate the intrinsic anemia in the adult and alter correct development in the early embryo. Even when expressed as a heterozygote, the Nan-EKLF protein accomplishes this by direct binding and aberrant activation of genes encoding secreted factors that exert a negative effect on erythropoiesis and iron use. Our data form the basis for a novel mechanism of physiological deficiency that is relevant to human dyserythropoietic anemia and likely other disease states.


Subject(s)
Anemia, Neonatal/genetics , Kruppel-Like Transcription Factors/genetics , Mutation , Amino Acid Substitution , Anemia, Neonatal/blood , Anemia, Neonatal/embryology , Animals , Animals, Newborn , Cytokines/blood , DNA/genetics , DNA/metabolism , Disease Models, Animal , Erythrocytes/metabolism , Erythropoiesis/genetics , Gene Expression Regulation, Developmental , Heterozygote , Humans , Kruppel-Like Transcription Factors/blood , Kruppel-Like Transcription Factors/deficiency , Mice , Mice, Knockout , Mice, Mutant Strains , Models, Biological , Muscle Proteins/blood , Mutant Proteins/blood , Mutant Proteins/genetics
20.
Transfusion ; 57(4): 1072-1077, 2017 04.
Article in English | MEDLINE | ID: mdl-28194794

ABSTRACT

BACKGROUND: In(Lu) is characterized by a reduced expression of antigens in the Lutheran blood group system as well as other blood group antigens. Mutations of the erythroid transcription factor, KLF1, have been reported to cause the In(Lu) phenotype, and we investigated Japanese In(Lu) to estimate the prevalence of the phenotype and KLF1 polymorphism. STUDY DESIGN AND METHODS: Blood samples were screened by monoclonal anti-CD44 and the In(Lu) phenotype was confirmed by tube tests including adsorption and elution tests using anti-Lua and anti-Lub . KLF1, LU, and A4GALT genes were analyzed by polymerase chain reaction and sequencing. RESULTS: We identified 100 of 481,322 blood donors (0.02%), and the previously characterized 20 donors, who had the In(Lu) phenotype with the LUB/LUB genotype. A total of 100 of the 120 In(Lu) individuals had mutant KLF1 alleles, and we identified 13 known and 21 novel alleles. The mutant KLF1 alleles with c.947G>A (p.Cys316Tyr), c.862A>G (p.Lys288Glu), or c.968C>G (p.Ser323Trp) were major in the In(Lu) individuals. The P1 antigen of 29 In(Lu) (two P1 /P1 , 27 P1 /P2 ) showed significantly weakened expression by hemagglutination. CONCLUSIONS: The prevalence of the In(Lu) phenotype in the Japanese population was 0.02%, and we identified 13 known and 21 novel KLF1 alleles. The KLF1 mutations cause the reduced expression of the P1 antigen.


Subject(s)
Cell Adhesion Molecules/genetics , Kruppel-Like Transcription Factors/genetics , Lutheran Blood-Group System/genetics , Mutation, Missense , Phenotype , Amino Acid Substitution , Asian People , Cell Adhesion Molecules/blood , Female , Galactosyltransferases/biosynthesis , Galactosyltransferases/genetics , Globosides/biosynthesis , Globosides/metabolism , Humans , Japan , Kruppel-Like Transcription Factors/blood , Lutheran Blood-Group System/blood , Male
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