RÉSUMÉ
Coronary artery disease (CAD) imposes a significant economic burden in developing countries like India. Timely diagnosis and treatment should be prioritized to mitigate the disease. Current diagnostic tools being invasive and less specific raise the need to develop less invasive and more reliable molecular biomarkers. MicroRNAs (miRNAs) are an emerging class of molecules that can serve as a potential source of non-invasive biomarkers for CAD. The objective of this study was to determine the potential of circulatory miRNAs as diagnostic biomarkers in CAD. In this study, we have reported two microRNAs, miR-128-3p and miR-195-5p in the serum of CAD patients in Indian Population. A total of 124 subjects were recruited which included 89 angiographically proven CAD patients and 35 control subjects. Our results show a significant decrease in the levels of miR-128-3p in CAD patients while there were no significant changes in the levels of miR-195-5p. Further bioinformatics analysis revealed the potential role of miR-128-3p in cholesterol homeostasis. Altered homeostasis due to cholesterol accumulation in macrophages is the driving force behind formation of foam cells which in turn accelerates the progression of CAD. Here, we have shown that miR-128-3p increases cholesterol levels in macrophages by decreasing cholesterol efflux in-vitro.
Sujet(s)
Marqueurs biologiques , Maladie des artères coronaires , microARN , Humains , microARN/génétique , microARN/sang , microARN/métabolisme , Maladie des artères coronaires/génétique , Maladie des artères coronaires/sang , Maladie des artères coronaires/métabolisme , Mâle , Femelle , Marqueurs biologiques/sang , Adulte d'âge moyen , Inde/épidémiologie , Projets pilotes , Études cas-témoins , Cholestérol/sang , Sujet âgé , AdulteRÉSUMÉ
Imbalance in glucose metabolism and insulin resistance are two primary features of type 2 diabetes/diabetes mellitus. Its etiology is linked to mitochondrial dysfunction in skeletal muscle tissue. The mitochondria are vital organelles involved in ATP synthesis and metabolism. The underlying biological pathways leading to mitochondrial dysfunction in type 2 diabetes can help us understand the pathophysiology of the disease. In this study, the mitochondrial gene expression dataset were retrieved from the GSE22309, GSE25462, and GSE18732 using Mitocarta 3.0, focusing specifically on genes that are associated with mitochondrial function in type 2 disease. Feature selection on the expression dataset of skeletal muscle tissue from 107 control patients and 70 type 2 diabetes patients using the XGBoost algorithm having the highest accuracy. For interpretation and analysis of results linked to the disease by examining the feature importance deduced from the model was done using SHAP (SHapley Additive exPlanations). Next, to comprehend the biological connections, study of protein-protien and mRNA-miRNA networks was conducted using String and Mienturnet respectively. The analysis revealed BDH1, YARS2, AKAP10, RARS2, MRPS31, were potential mitochondrial target genes among the other twenty genes. These genes are mainly involved in the transport and organization of mitochondria, regulation of its membrane potential, and intrinsic apoptotic signaling etc. mRNA-miRNA interaction network revealed a significant role of miR-375; miR-30a-5p; miR-16-5p; miR-129-5p; miR-1229-3p; and miR-1224-3p; in the regulation of mitochondrial function exhibited strong associations with type 2 diabetes. These results might aid in the creation of novel targets for therapy and type 2 diabetes biomarkers.
Sujet(s)
Diabète de type 2 , microARN , Maladies mitochondriales , Humains , Diabète de type 2/génétique , Diabète de type 2/métabolisme , Intelligence artificielle , microARN/génétique , microARN/métabolisme , Muscles squelettiques/métabolisme , Mitochondries/génétique , Mitochondries/métabolisme , ARN messager/génétique , Maladies mitochondriales/métabolisme , Apprentissage machineRÉSUMÉ
BACKGROUND: Computational tools may have an edge over conventional methods for the preliminary evaluation of food allergenicity. In this study, the allergenic potential of Lentinula edodes was evaluated and validated using in silico tools. RESULTS: The potential cross-reactivity of mushroom proteins with fungal allergens was determined using sequence alignment - the Fast Alignment (FASTA) and Basic Local Alignment Search Tool (BLAST) algorithm. Eight L. edodes proteins were cross-reactive with allergens from fungal origin, showing 52%-89% sequence identity using FASTA algorithm-based alignment. The BLAST data were corroborated by percentage identity and query coverage. Physico-chemical property-based allergenicity was deciphered by AlgPred, Allermatch, and AllergenFP software, which predicted six out of eight proteins as potential allergens. Sequence alignment showed 66%-86% conservancy between mushroom protein and known fungal allergens. Secondary structure and amino acid composition supported structural affinity between query and fungal proteins. Three-dimensional structures of five mushroom proteins were generated, quality assessed, and superimposed with fungal allergens, suggesting possible allergenicity of mushroom proteins. An enzyme-linked immunosorbent assay (ELISA) demonstrated immunoglobulin E (IgE) binding in 13 out of 21 food-hypersensitive patients' sera. CONCLUSION: In silico tools provide preliminary indications about the potential allergenicity and cross-reactivity of mushroom proteins. This approach may be used for the prelusive allergenicity assessment of allergen sources. © 2022 Society of Chemical Industry.
Sujet(s)
Hypersensibilité alimentaire , Champignons shiitake , Humains , Allergènes/composition chimique , Alignement de séquences , Réactions croiséesRÉSUMÉ
Targeted therapy is receiving considerable attention from the researchers around the globe owing to the increased drug-resistance and incidences of cancer recurrences. MicroRNAs (miRNAs) exhibits tremendous potential as a candidate for molecular targeted therapy in cancer. Unfortunately, majority of research related to microRNAs are focussed on either a particular miRNA or a set of unrelated miRNAs. There is lack of holistic knowledge on differential co-expression of miRNA clusters in regulating the gene expression under physiological conditions. Previously, we reported the cooperative effect of hsa-miR-23a~27a~24-2 cluster in inducing ER (Endoplasmic Reticulum) stress-mediated apoptotic cell death of HEK cells. In the present study, we have investigated the common anti-cancer effects of individual members of this cluster. Our in silico analysis identified twelve common target genes distributed across three independent clusters. Furthermore, we found NCOA1, NLK, and RAP1B to fall in a single cluster with NCOA1 as a central hub molecule. Prognostic analysis showed profound involvement of these three genes in the breast cancer progression and metastasis. We further demonstrated that alteration in the levels of individual members of miR-23a~27a~24-2 cluster commonly regulates the invasive migration of breast cancer cells by modulating EMT and cytoskeletal pathway proteins. Our results reveal a new insight into the therapeutic potential of individual members of the pro-apoptotic hsa-miR-23a~27a~24-2 cluster family against metastatic breast cancer.
Sujet(s)
Tumeurs du sein , microARN , Apoptose/génétique , Tumeurs du sein/génétique , Tumeurs du sein/anatomopathologie , Stress du réticulum endoplasmique , Femelle , Humains , microARN/métabolisme , Récidive tumorale locale , Coactivateur-1 de récepteur nucléaire , Protein-Serine-Threonine Kinases , Protéines G rap/métabolismeRÉSUMÉ
The size and morphology of mitochondria are very heterogeneous and correlates well with their healthy functioning. In many pathological conditions, mitochondrial morphology is altered due to impaired mitochondrial dynamics (a collective term for mitochondrial fusion and fission) and dysfunction. The current study aimed at identifying the role of microRNA-128 (miR-128) in regulating mitochondrial biogenesis. Previously, peroxisome proliferator activator receptor γ coactivator 1α (PGC1α) has been shown to co-activate key intermediates of mitochondrial biogenesis, function, and dynamics; however, the upstream regulatory network remains largely unknown. We, herein using in silico analysis followed by in vitro experiments in C2C12 myoblasts, showed that miR-128 reduces mitochondrial biogenesis by directly targeting PGC1α. The expression of downstream genes, nuclear respiratory factors 1 and 2 (NRF1 and NRF2, respectively), and mitochondrial transcription factor A (TFAM) were decreased in C2C12 myoblasts upon overexpression of miR-128. Also, miR-128 is shown to promote mitochondrial dysfunction by directly targeting NADH Dehydrogenase (Ubiquinone) Fe-S Protein 4 (NDUFS4). The mitochondrial dynamics and morphology were impaired post miR-128 overexpression, as revealed by downregulation of fusion proteins (mitofusin1 and 2, i.e., MFN1 and MFN2, respectively) and upregulation of fission protein (dynamin-related protein 1, i.e., DRP1). Conversely, inhibition of miR-128 expression improved mitochondrial biogenesis, function, and dynamics, as evidenced by increased mitochondrial mass and ATP production after antimiR-128 treatment. Our findings reveal that inhibition of miR-128 can be a new potential target for reversing the effects of metabolic disorders of skeletal muscle as observed during many pathophysiological conditions such as obesity and type II diabetes.
Sujet(s)
Complexe I de la chaîne respiratoire/métabolisme , microARN/métabolisme , Mitochondries du muscle/métabolisme , Myoblastes/métabolisme , Coactivateur 1-alpha du récepteur gamma activé par les proliférateurs de peroxysomes/métabolisme , Animaux , Lignée cellulaire , Complexe I de la chaîne respiratoire/génétique , Régulation de l'expression des gènes , Mâle , Souris , Souris de lignée C57BL , microARN/génétique , Mitochondries du muscle/génétique , Consommation d'oxygène , Coactivateur 1-alpha du récepteur gamma activé par les proliférateurs de peroxysomes/génétiqueRÉSUMÉ
Mitochondria are not only important for cellular bioenergetics but also lie at the heart of critical metabolic pathways. They can rapidly adjust themselves in response to changing conditions and the metabolic needs of the cell. Mitochondrial involvement as well as its dysfunction has been found to be associated with variety of pathological processes and diseases. mitomiRs are class of miRNA(s) that regulate mitochondrial gene expression and function. This review sheds light on the role of mitomiRs in regulating different biological processes-mitochondrial dynamics, oxidative stress, cell metabolism, chemoresistance, apoptosis,and their relevance in metabolic diseases, neurodegenerative disorders, and cancer. Insilico analysis of predicted targets of mitomiRs targeting energy metabolism identified several significantly altered pathways (needs in vivo validations) that may provide a new therapeutic approach for the treatment of human diseases. Last part of the review discusses about the clinical aspects of miRNA(s) and mitomiRs in Medicine.
Sujet(s)
microARN/génétique , Mitochondries/génétique , Maladies mitochondriales/génétique , Tumeurs/génétique , Apoptose/génétique , Métabolisme énergétique/génétique , Régulation de l'expression des gènes/génétique , Humains , Mitochondries/métabolisme , Mitochondries/anatomopathologie , Maladies mitochondriales/métabolisme , Maladies mitochondriales/anatomopathologie , Tumeurs/métabolisme , Tumeurs/anatomopathologie , Stress oxydatif/génétiqueRÉSUMÉ
AIMS: Hypercholesterolemia remains a critical risk factor for cardiovascular diseases and there is an urgent need to develop effective alternative therapeutics. Herein, we investigated the effects of miR-128-3p inhibition on serum cholesterol levels using a hypercholesterolemic mouse model. MATERIALS AND METHODS: Five injections of anti-miR-128-3p (AM-128) treatment were given, and the cholesterol profile in serum and liver was quantified. We validated the underlying gene network using qRT-PCR, western blotting, ELISA, and dual luciferase assays. KEY FINDINGS: AM-128 treatment inhibits cholesterol biosynthesis by upregulating INSIG1 and downregulating HMGCR (3-hydroxy-3-methylglutaryl-CoA reductase) expression. The serum cholesterol clearance by SR-B1 (scavenger receptor class B member 1) and LDLR (low density lipoprotein receptors) was also increased. Furthermore, the catabolism of cholesterol by CYP7A1 (cytochrome P450 family 7 subfamily A member 1) was increased. SIGNIFICANCE: Our results confirmed a critical role of miR-128-3p inhibition in lowering serum cholesterol and suggest its potential therapeutic implications in reversing hypercholesterolemia.
Sujet(s)
Hypercholestérolémie/génétique , microARN/génétique , Animaux , Maladies cardiovasculaires/prévention et contrôle , Lignée cellulaire tumorale , Cholestérol/sang , Cholestérol/métabolisme , Cholesterol 7-alpha-hydroxylase/métabolisme , Modèles animaux de maladie humaine , Fibrose/métabolisme , Cellules HepG2 , Humains , Hypercholestérolémie/métabolisme , Hypercholestérolémie/thérapie , Interféron gamma/métabolisme , Protéines et peptides de signalisation intracellulaire/métabolisme , Lipides/composition chimique , Foie/métabolisme , Mâle , Protéines membranaires/métabolisme , Souris , Souris de lignée C57BL , microARN/antagonistes et inhibiteurs , Facteurs de risqueRÉSUMÉ
Hypobaric hypoxia poses stress to sojourners traveling to high-altitude. A cascade of physiological changes occurs to cope with or adapt to hypobaric hypoxia. However, an insufficient physiological response to the hypoxic condition resulting from imbalanced vascular homeostasis pathways results in high-altitude pulmonary edema (HAPE). The present study aims to identify the implication of miRNAs associating with HAPE and adaptation. We analyzed the expression of 1,113 miRNAs in HAPE-patients (HAPE-p), HAPE-free controls (HAPE-f), and highland natives (HLs). Based on miRNA profiling and in silico analyses, miR-124-3p emerged relevantly. We observed a significant overexpression of miR-124-3p in HAPE-p. In silico analyses revealed a direct interaction of miR-124-3p with vascular homeostasis and hypoxia-associated genes NOS3 (endothelial nitric oxide synthase), Apelin, and ETS1 (V-Ets avian erythroblastosis virus E2 oncogene homolog 1). Moreover, the transcript and biolevel expression of these genes were significantly decreased in HAPE-p when compared with HAPE-f or HLs. Our in vitro analysis in human umbilical vein endothelial cells demonstrated a significant knockdown of these genes both at transcript and protein levels following miR-124-3p overexpression. Conclusively, our results showed that miR-124-3p might play a plausible role in HAPE pathophysiology by inhibiting the expression of NOS3, Apelin, and ETS1.
Sujet(s)
Mal de l'altitude/sang , Mal de l'altitude/métabolisme , Hypertension pulmonaire/sang , Hypertension pulmonaire/métabolisme , Hypoxie/sang , Hypoxie/métabolisme , microARN/sang , Oedème pulmonaire/sang , Oedème pulmonaire/métabolisme , Adaptation physiologique/physiologie , Adulte , Altitude , Apeline/métabolisme , Lignée cellulaire , Femelle , Cellules HEK293 , Cellules endothéliales de la veine ombilicale humaine , Humains , Mâle , Nitric oxide synthase type III/métabolisme , Protéine proto-oncogène c-ets-1/métabolisme , Jeune adulteRÉSUMÉ
INTRODUCTION: With the advent and extensive use of the Internet and smartphones, social networking has become a pervasive part of human interaction. The use of these social networking sites or the Internet affects the physical, mental, and spiritual health of the people. Hence, there is need to understand how the time spent on social networking is affecting the quality of life (QOL) as a whole, especially among college-going students who are most likely users of social networking sites (18-21 years). MATERIALS AND METHODS: A cross-sectional survey was conducted among 220 college-going students (18-21 years) in Chandigarh in 2012. The data were collected using a pretested self-administered questionnaire, adapted from Young's Internet usage questionnaire. Appropriate statistical analysis was done. RESULTS: Almost all (98%) of the respondents use the Internet. As compared to nondaily users of social networking sites, daily users were better able to handle stress related to (1) relationships (moderate to severe stress among daily users vs. nondaily users, 15.2% vs. 30.5%) and (2) work (moderate to severe stress among daily users vs. nondaily users, 18.2% vs. 35.4%). The daily users of social networking sites feel significantly more satisfied with their classmates, the way they handle the problems, their physical appearance, and their accomplishments in their life. CONCLUSION: Social networking sites are steadily penetrating in the lives of adolescents in India. The advantages on quality of life for daily users of social networking sites versus nondaily users are enormous. Also currently, Internet use might not have reached the levels where it embarks on the existing state of health; therefore, continuous and critical observation of the changing trends is warranted.
Sujet(s)
Qualité de vie , Réseautage social , Étudiants , Universités , Adolescent , Adulte , Études transversales , Exercice physique , Femelle , Humains , Mâle , Santé mentale , Stress psychologique , Enquêtes et questionnaires , Jeune adulteRÉSUMÉ
PURPOSE: Non-canonical NFκB (NC-NFκB) pathway plays an influential role in metastasis, which promotes cancer proliferation and progression. The aim of the study was to examine the expression of NC-NFκB proteins and their correlation with clinicopathological factors associated with metastatic cases of uveal melanoma (UM) and with the patient outcome. METHOD: Expression of NC-NFκB proteins (p52, RelB, and co-expression of p52/RelB) was evaluated in 75 formalin-fixed cases of uveal melanoma by immunohistochemistry. Validation of nuclear immunoreactivity was done by western blotting. Transcriptional status of NC-NFκB genes was assessed in 60 fresh tumor tissues by quantitative real-time PCR. Co-immunoprecipitation was performed to determine the presence of native p52/RelB heterodimer in UM. Prognostic relevance was determined using Cox proportional hazard and Kaplan-Meier methods. RESULTS: Immunohistochemical expression of p52, RelB, and their co-expression was observed in 81%, 68.7%, 56.2% of metastatic cases, respectively, while their expression was seen only in 38%, 33% and 30% of non-metastatic cases. Loss of BAP-1 was correlated with expression of p52 and RelB proteins. Co-immunoprecipitation assay confirmed the putative interaction of p52 with RelB protein in metastatic cases of uveal melanoma. Co-expression of p52/RelB and expression of p52 protein was significantly correlated with decreased metastasis-free survival (MFS) (p = 0.004; p = 0.002) and overall survival (OS) (p = 0.004; p = 0.032), while the RelB expression only correlated with reduced MFS (p = 0.003). CONCLUSION: Our data showed that non-canonical NFκB proteins were significantly higher in metastatic cases and associated with poor outcome of the patients. Furthermore, the p52 protein could be used as a potential therapeutic biomarker for metastatic cases in uveal melanoma.
Sujet(s)
Régulation de l'expression des gènes tumoraux/génétique , Mélanome/génétique , Sous-unité p52 de NF-kappa B/génétique , Métastase tumorale/génétique , Facteur de transcription RelB/génétique , Tumeurs de l'uvée/génétique , Adulte , Survie sans rechute , Femelle , Humains , Immunohistochimie/méthodes , Mâle , Mélanome/anatomopathologie , Métastase tumorale/anatomopathologie , Pronostic , Études prospectives , Transcription génétique/génétique , Tumeurs de l'uvée/anatomopathologieRÉSUMÉ
Inflammation in uveal melanoma (UM) is linked to a bad prognosis. It is rare type of cancer, of which the metastases are usually fatal within a year. Infiltration with an inflammatory infiltrate increases with disease progression but does not seem to inhibit metastasis. The Canonical NFκB (C-NFκB) pathway is known to play a crucial role in tumor inflammation. We therefore, studied the expression of canonical NFκB proteins and their prognostic relevance in UM. Our study evaluated the expression of C-NFκB proteins (p65, p50, and c-Rel) by using immunohistochemistry on sections from 75 formalin-fixed UM. Activation of the NFκB subunit was determined on fresh tumor specimens by measuring the DNA-binding activity in nuclei using an NFκB ELISA assay. Real-time PCR was performed on frozen material on 58 tumors. The presence of native C-NFκB heterodimers (p65/p50 and c-Rel/p50) was confirmed by co-immunoprecipitation followed by Western blotting. We observed a high nuclear immunoreactivity of p65, p50, and c-Rel proteins in 54, 60 and 41% UM cases, respectively. Expression of C-NFκB proteins significantly correlated with parameters which are related to the inflammatory environment of UM. Nuclear immunoreactivity of p65 and p50 was associated with lower patient survival (p = 0.041; p = 0.048) while c-Rel was not. Our finding reveals that C-NFκB proteins expressed are more often in UM with inflammation than those without inflammation. Activation of the canonical NFκB pathway is more frequent in high risk UM patients. These observations might help to understand the behaviour of high risk tumors, with upregulation of C-NFκB proteins contributing to tumor aggressiveness.
Sujet(s)
Mélanome/anatomopathologie , Sous-unité p50 de NF-kappa B/métabolisme , Protéines proto-oncogènes c-rel/métabolisme , Facteur de transcription RelA/métabolisme , Tumeurs de l'uvée/anatomopathologie , Protéines de liaison à l'ADN/métabolisme , Humains , Mélanocytes/anatomopathologie , Mélanome/mortalité , Uvée/anatomopathologie , Tumeurs de l'uvée/mortalitéRÉSUMÉ
Mitochondrial dynamics is a highly dysregulated process in cancer. Apoptosis and mitochondrial fission are two concurrent events wherein increased mitochondrial fragmentation serves as a hallmark of apoptosis. We have shown earlier that miR-195 exerts pro-apoptotic effects in breast cancer cells. Herein, we have demonstrated miR-195 as a modulator of mitochondrial dynamics and function. Imaging experiments upon miR-195 treatment have shown that mitochondria undergo extensive fission. We validated mitofusin2 as a potential target of miR-195. This may provide a molecular explanation for the respiratory defects induced by miR-195 over-expression in breast cancer cells. Active, but not total, mitochondrial mass, was reduced with increasing levels of miR-195. We have further shown that miR-195 enhances mitochondrial SOD-2 expression but does not affect PINK1 levels in breast cancer cells. Collectively, we have revealed that miR-195 is a modulator of mitochondrial dynamics by targeting MFN2 thereby impairing mitochondrial function. Concomitantly, it enhances the scavenger of reactive oxygen species (SOD-2) to maintain moderate levels of oxidative stress. Our findings suggest a therapeutic potential of miR-195 in both ER-positive as well as ER-negative breast cancer cells.
Sujet(s)
Tumeurs du sein/génétique , dGTPases/métabolisme , microARN/métabolisme , Mitochondries/métabolisme , Protéines mitochondriales/métabolisme , Régions 3' non traduites/génétique , Apoptose , Séquence nucléotidique , Lignée cellulaire tumorale , Respiration cellulaire , Régulation négative/génétique , Femelle , Régulation de l'expression des gènes tumoraux , Humains , Mitochondries/ultrastructure , Dynamique mitochondriale , Mitophagie , Stress oxydatif , Consommation d'oxygèneRÉSUMÉ
Alteration in mitochondrial DNA plays an important role in the development and progression of cancer. The Displacement Loop (D-loop) region of mitochondrial DNA (mtDNA) is the regulatory region for its replication and transcription. Therefore, we aimed to characterize mutations in the D-loop region of mitochondrial DNA along with the morphological changes and analyzed their impact on survival in retinoblastoma patients. mtDNA D-loop region was amplified by Nested-Polymerase Chain Reaction (Nested-PCR) and mutations were analyzed in 60 tumor samples from retinoblastoma patients by DNA sequencing. Transmission electron microscopy was performed on 5 retinoblastoma specimens. Mutations were correlated with clinical, histopathological parameters and patient survival. D-loop mutations were found in total of 52/60 (86.6%) patients. The most common mutations were T to C and C to T followed by A to G. There were 5.81% mutations which were not previously reported in the MITOMAP database. A73G (83.33%) were the most frequent mutations found in our cases and it was statistically significant with poor tumor differentiation and age. In addition, this study was further analyzed for morphological changes in retinoblastoma that had disorganized, swollen and less numbers of mitochondria on electron microscopy. This is the first study showing high frequency of mtDNA mutation which might be due to abnormal morphology of mitochondria in retinoblastoma. Our results indicate that pathogenic mtDNA D-loop mutations may be involved in tumorigenesis of retinoblastoma tumor.
Sujet(s)
ADN mitochondrial/génétique , Tumeurs de la rétine/génétique , Tumeurs de la rétine/anatomopathologie , Rétinoblastome/génétique , Rétinoblastome/anatomopathologie , Enfant d'âge préscolaire , Analyse de mutations d'ADN , Femelle , Humains , Mâle , MutationRÉSUMÉ
Psoriasis is an immune-mediated inflammatory disease of the skin. Previous studies including ours have shown that IL-17A plays a major role in its pathogenesis; however, its precise molecular mechanism of action is not well understood. Cytokines like TNF α and IL-23 are also important in mediating the disease and some studies have also reported autophagy as a novel mechanism by which cytokines controls the immune response. Herein, we investigated the effect of IL-17A on autophagy and reveal crosstalk between autophagy and cholesterol signaling in keratinocytes. Our results suggest that IL-17A stimulated keratinocytes activated PI3K/AKT/mTOR signaling and inhibited autophagy by simultaneously inhibiting autophagosome formation and enhancing autophagic flux. Western blotting was utilized to detect the expression of autophagic markers (LC3 and p62), PI3K, mTOR and AKT. Induction of autophagy by mTOR inhibitor rapamycin and/or starvation also inhibited the levels of IL-17A secreted IL-8, CCL20 and S100A7 in keratinocytes. Herein, we also observed that inhibition of autophagy by IL-17A was accompanied by enhanced cellular cholesterol levels which in turn regulated the autophagic flux. To investigate crosstalk between autophagy and cellular cholesterol, we used methyl-ß-cyclodextrin (MßCD), which disrupts detergent-insoluble microdomains (DIMs) by depleting cells of cholesterol and checked autophagy. Decreased expression of LC3-II in psoriatic lesional skin compared to non-lesional skin and induction of autophagy by anti-psoriatic drug methotrexate in keratinocytes further confirms the role of autophagy in psoriasis. Our findings suggest that modulators of autophagy and/or cholesterol levels may be developed, and also may lead to new therapeutic agents for psoriasis treatment.
Sujet(s)
Autophagie , Cholestérol/métabolisme , Interleukine-17/métabolisme , Kératinocytes/métabolisme , Phosphatidylinositol 3-kinases/métabolisme , Protéines proto-oncogènes c-akt/métabolisme , Psoriasis/métabolisme , Sérine-thréonine kinases TOR/métabolisme , Lignée cellulaire , Cholestérol/génétique , Humains , Inflammation/génétique , Inflammation/métabolisme , Inflammation/anatomopathologie , Interleukine-17/génétique , Kératinocytes/anatomopathologie , Phosphatidylinositol 3-kinases/génétique , Protéines proto-oncogènes c-akt/génétique , Psoriasis/génétique , Psoriasis/anatomopathologie , Sérine-thréonine kinases TOR/génétiqueRÉSUMÉ
Psoriasis is recognized as a T cell mediated inflammatory hyperproliferative skin disorder. Several microRNAs (miRNAs) have been implicated in the pathogenesis of psoriasis however, understanding of their mechanistic involvement remains unclear. Previously, we have shown that PUVA induced miR-4516 downregulates signal transducer and activator of transcription 3 (STAT3) by direct binding to its 3' untranslated region (3'UTR) and suppresses STAT3 downstream genes (Bcl xl, Cyclin D1). Here, we demonstrate for the first time that expression of miR-4516 is significantly downregulated in psoriatic skin. We additionally validated extracellular matrix protein fibronectin 1 (FN1) and integrin subunit α9 (ITGA9) as direct targets of miR-4516. Interestingly, ITGA9 expression was found to be increased in the suprabasal psoriatic epidermis. We further showed that ectopic expression of miR-4516 in human keratinocytes not only suppresses cell motility and proliferation via significant downregulation of genes orchestrating cytoskeletal reorganization (Rac1, RhoA, Cdc42), but also inhibits F-actin assembly and induces terminal differentiation. Collectively, our results provide evidence that loss of expression of miR-4516 in psoriatic skin might be contributing to accelerated migration, resistance to apoptosis and differentiation as seen in psoriasis lesional keratinocytes and also highlight its potential as a novel small molecule for therapeutic intervention in psoriasis.
Sujet(s)
Fibronectines/métabolisme , Intégrines alpha/métabolisme , microARN/métabolisme , Psoriasis/métabolisme , Régions 3' non traduites , Actines/métabolisme , Différenciation cellulaire/physiologie , Lignée cellulaire , Mouvement cellulaire/génétique , Prolifération cellulaire/physiologie , Régulation négative , Fibronectines/génétique , Humains , Kératinocytes/métabolisme , Kératinocytes/anatomopathologie , microARN/génétique , Psoriasis/génétique , Facteur de transcription STAT-3/génétique , Facteur de transcription STAT-3/métabolisme , Transduction du signal , Protéine G cdc42/génétique , Protéine G cdc42/métabolisme , Protéine G rac1/génétique , Protéine G rac1/métabolisme , Protéine G RhoA/génétique , Protéine G RhoA/métabolismeRÉSUMÉ
Previously, we reported significantly decreased expression of tissue and circulating miR-107 in esophageal cancer (EC). However, its role in esophageal tumorigenesis still remains elusive. Therefore, the aim of the present study was to analyze the role of miR-107 in esophageal squamous cell carcinoma (ESCC). The role of miR-107 in ESCC was evaluated using MTT assay, cell cycle analysis by flow cytometry, annexin assay, colony formation assay and scratch assay. Overexpression of miR-107 in KYSE-410 cells suppressed cell proliferation at 72 h post-transfection (p=0.0001). Moreover, a significant increase in the G0/G1 population (p<0.001) and a significant decrease in the G2/M (p=0.032) population was also observed in the miR-107-treated cells as compared to the negative control (NC). Notably, miR-107 overexpression attenuated the colony formation potential of ESCC cells by 41.83% as compared to the NC (p=0.007). miR-107 mimic inhibited ESCC cell migration in a time-dependent manner, reducing the wound closure to only 50.41±7.23% at 72 h post-transfection (p=0.041). Further analysis by Matrigel invasion assay revealed a significant decrease in the migratory and invasive abilities of the KYSE-410 cells at 72 h post miR-107 transfection. qRT-PCR analysis showed decreased expression of one of the newly identified targets of miR-107, Cdc42, at the mRNA level. Further validation by western blotting confirmed a significant reduction in the identified target at the protein level. In addition, the relative luciferase activity of the reporter containing Cdc42 3'UTR was significantly decreased upon miR-107 co-transfection, indicating it to be a direct target of miR-107. Our results herein document that miR-107 functions as a tumor suppressor and inhibits the proliferation, migration and invasion of ESCC cells. Moreover, this is the first report showing Cdc42 as a downstream target of miR-107.
Sujet(s)
Carcinome épidermoïde/génétique , Régulation négative , Tumeurs de l'oesophage/génétique , microARN/génétique , Protéine G cdc42/génétique , Régions 3' non traduites , Apoptose , Lignée cellulaire tumorale , Mouvement cellulaire , Prolifération cellulaire , Carcinome épidermoïde de l'oesophage , Régulation de l'expression des gènes tumoraux , Humains , Invasion tumorale , Régions promotrices (génétique)RÉSUMÉ
Lipid rafts are dynamic assemblies of proteins and lipids that harbour many receptors and regulatory molecules and so act as a platform for signal transduction. They float freely within the liquid-disordered bilayer of cellular membranes and can cluster to form larger ordered domains. Alterations in lipid rafts are commonly found to be associated with the pathogenesis of several human diseases and recent reports have shown that the raft domains can also be perturbed by targeting raft proteins through microRNAs. Over the last few years, the importance of lipid rafts in modulating both innate and acquired immune responses has been elucidated. Various receptors present on immune cells like B cells, T cells, basophils and mast cells associate with lipid rafts on ligand binding and initiate signalling cascades leading to inflammation. Furthermore, disrupting lipid raft integrity alters lipopolysaccharide-induced cytokine secretion, IgE signalling, and B-cell and T-cell activation. The objective of this review is to summarize the recent progress in understanding the role of lipid rafts in the modulation of immune signalling and its related therapeutic potential for autoimmune diseases and inflammatory disorders.
Sujet(s)
Maladies auto-immunes/thérapie , Lymphocytes B/physiologie , Immunothérapie/méthodes , Microdomaines membranaires/métabolisme , microARN/génétique , Transduction du signal/immunologie , Lymphocytes T/physiologie , Animaux , Maladies auto-immunes/immunologie , Cytokines/génétique , Cytokines/métabolisme , Humains , Immunoglobuline E/génétique , Immunoglobuline E/métabolisme , Immunothérapie/tendances , Activation des lymphocytesRÉSUMÉ
Resistance to apoptosis leads to tumorigenesis and failure of anti-cancer therapy. Recent studies also highlight abrogated lipid/cholesterol metabolism as one of the root causes of cancer that can lead to metastatic transformations. Cancer cells are dependent on tremendous supply of cellular cholesterol for the formation of new membranes and continuation of cell signaling. Cholesterol homeostasis network tightly regulates this metabolic need of cancer cells on cholesterol and other lipids. Genetic landscape is also shared between apoptosis and cholesterol metabolism. MicroRNAs (miRNAs) are the new fine tuners of signaling pathways and cellular processes and are known for their ability to post-transcriptionally repress gene expression in a targeted manner. This review summarizes the current knowledge about the cross talk between apoptosis and cholesterol metabolism via miRNAs. In addition, we also emphasize herein recent therapeutic modulations of specific miRNAs and their promising potential for the treatment of deadly diseases including cancer and cholesterol related pathologies. Understanding of the impact of miRNA-based regulation of apoptosis and metabolic processes is still at its dawn and needs further research for the development of future miRNA-based therapies. As both these physiological processes affect cellular homeostasis, we believe that this comprehensive summary of miRNAs modulating both apoptosis and cholesterol metabolism will open uncharted territory for scientific exploration and will provide the foundation for discovering novel drug targets for cancer and metabolic diseases.
Sujet(s)
Apoptose , Cholestérol/métabolisme , Maladies métaboliques/génétique , Maladies métaboliques/anatomopathologie , microARN/génétique , Tumeurs/génétique , Tumeurs/anatomopathologie , Animaux , HumainsRÉSUMÉ
BACKGROUND: Reactive oxygen species (ROS) have been shown to enhance the proliferation of cancer cells. NADPH oxidases (NOX4) are a major intracellular source of ROS and are found to be associated with cancer and tumor cell invasion. Therefore, the purpose of this study is to evaluate the expression of NOX4 protein in human retinoblastoma. METHODS: Immunohistochemical expression of NOX4 protein was analyzed in 109 specimens from prospective cases of retinoblastoma and then correlated with clinicopathological parameters and patient survival. Western blotting confirmed and validated the immunoreactivity of NOX4 protein. RESULTS: In our study we found a male preponderance (55.9 %), and 25/109 (22.9 %) were bilateral. Massive choroidal invasion was the histopathological high-risk factor (HRF) most frequently observed, in 42.2 % of the cases. NOX4 protein was expressed in 67.88 % (74/109) of primary retinoblastoma cases and was confirmed by Western blotting. NOX4 was statistically significant with massive choroidal invasion and pathological TNM staging. There was a statistically significant difference in overall survival in patients with NOX4 expression (p = 0.0461). CONCLUSION: This is the first study to show the expression of NOX4 protein in retinoblastoma tumors. Hence, a retinoblastoma tumor may exhibit greater ROS stress. This protein may prove to be useful as a future therapeutic target for improving the management of retinoblastoma.