Nucleolar RNA optical density (concentration) measurements at the single cell level indicated that differentiation of lymphocytes is accompanied by a slightly decreased nucleolar RNA concentration in contrast to the cytoplasmic rim around the nucleus. On the other hand, the nucleolar size was markedly reduced and the cytoplasmic rim surrounding the nucleus was reduced only weakly. Concerning the calculated rough estimate of the RNA content, the differentiation induced its larger decrease in the nucleoli than in the cytoplasmic rim. These observations indicated that the nucleolar RNA concentration and RNA content together with the nucleolar morphology are more sensitive markers of the differentiation process than the RNA concentration and content in the cytoplasm. Thus, the nucleolar RNA transfer to the cytoplasm in advanced differentiation steps might still be going on regardless of the decreasing or inhibited nucleolar biosynthetic activity. In addition, the presence of ring-shaped nucleoli and micronucleoli characteristic of mature and terminal lymphocytes in some lymphocytic less differentiated steps, i.e., lymphoblasts and prolymphocytes, might indicate the premature differentiation state of such cells.
- MeSH
- buněčné jadérko MeSH
- chronická lymfatická leukemie * MeSH
- cytoplazma MeSH
- lidé MeSH
- lymfocyty MeSH
- RNA MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Adjuvant therapy and radiotherapy improves the survival of patients with metastatic and locally advanced gastric cancer (GC). However, the resistance to radiotherapy limits its clinical usage. Rhotekin 2 (RTKN2) functions as an oncogene and confers resistance to ultraviolet B-radiation and apoptosis- inducing agents. Here, the role of RTKN2 in radiosensitivity of GC cell lines was investigated. RTKN2 was found to be elevated in GC tissues and cells. A series of functional assays revealed that overexpression of RTKN2 induced GC cell proliferation, promoted GC cell migration and invasion, while inhibiting GC cell apoptosis. However, silence of RTKN2 promoted GC cell apoptosis, while repressing GC cell proliferation, invasion and migration. GC cells were exposed to irradiation, and data from cell survival and apoptotic assays showed that knock-down of RTKN2 enhanced radiosensitivity of GC through up-regulation of apoptosis and down-regulation of proliferation in irradiation-exposed GC cells. Moreover, the protein expression of β-catenin and c-Myc in GC cells was enhanced by RTKN2 over-expression, but reduced by RTKN2 silence. Interference of RTKN2 down-regulated nuclear β-catenin expression, while up-regulating cytoplasmic β-catenin in GC. In conclusion, RTKN2 contributed to cell growth and radioresistance in GC through activation of Wnt/β-catenin signalling.
- MeSH
- beta-katenin metabolismus MeSH
- intracelulární signální peptidy a proteiny metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory žaludku * genetika patologie radioterapie MeSH
- pohyb buněk MeSH
- proliferace buněk MeSH
- regulace genové exprese u nádorů MeSH
- signální dráha Wnt MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Proliferation and migration of retinal endothelial cells (RECs) contribute to the development of diabetic retinopathy. PLAG1 (pleomorphic adenoma gene 1) functions as a zinc-finger transcription factor to participate in the development of lipoblastomas or pleomorphic adenomas of the salivary glands through regulation of cell proliferation and migration. The role of PLAG1 in diabetic retinopathy was investigated in this study. Firstly, RECs were induced under high glucose conditions, which caused reduction in viability and induction of apoptosis in the RECs. Indeed, PLAG1 was elevated in high glucosetreated RECs. Functional assays showed that silence of PLAG1 increased viability and suppressed apoptosis in high glucose-induced RECs, accompanied with up-regulation of Bcl-2 and down-regulation of Bax and cleaved caspase-3. Moreover, migration of RECs was promoted by high glucose conditions, while repressed by knockdown of PLAG1. High glucose also triggered angiogenesis of RECs through up-regulation of vascular endothelial growth factor (VEGF). However, interference of PLAG1 reduced VEGF expression to retard the angiogenesis. Silence of PLAG1 also attenuated high glucose-induced up-regulation of Wnt3a, β-catenin and c-Myc in RECs. Moreover, silence of PLAG1 ameliorated histopathological changes in the retina of STZ-induced diabetic rats through down-regulation of β-catenin. In conclusion, knockdown of PLAG1 suppressed high glucose-induced angiogenesis and migration of RECs, and attenuated diabetic retinopathy by inactivation of Wnt/ β-catenin signalling.
- MeSH
- beta-katenin metabolismus MeSH
- diabetická retinopatie * MeSH
- DNA vazebné proteiny metabolismus MeSH
- endoteliální buňky metabolismus MeSH
- experimentální diabetes mellitus * MeSH
- glukosa toxicita MeSH
- kaspasa 3 MeSH
- krysa rodu rattus MeSH
- patologická angiogeneze MeSH
- protein X asociovaný s bcl-2 MeSH
- retina metabolismus MeSH
- signální dráha Wnt MeSH
- transkripční faktory MeSH
- vaskulární endoteliální růstový faktor A MeSH
- zinek MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Non-small cell lung cancer (NSCLC) results in high mortality and has gained increasing attention. C-Phycocyanin (C-PC) has been identified as a potential therapeutic inhibitor for NSCLC, but its underlying mechanism remains obscure. The gene expression of the long noncoding RNA neighbour of BRCAI RNA 2 (NBR2) in NSCLC cells was evaluated by quantitative reverse transcription-PCR. The cell capacity for proliferation and migration was examined by EdU and wound-healing assays. Furthermore, the viability and apoptosis of cells was measured with CCK-8 and annexin V/PI, respectively. Next, the protein level of activation of adenosine monophosphate- activated protein kinase and the rapamycin kinase (mTOR) signalling pathway-associated molecules was evaluated by western blotting. H292 cells were pre-treated with C-PC or transfected with plasmids encoding NBR2 or the shNBR2 plasmid, to over-express or knock down NBR2 expression, respectively. NBR2 expression was robustly down-regulated in NSCLC cell lines compared with a normal cell line (BEAS-2B). NBR2 over-expression inhibited migration and promoted apoptosis of H292 cells. Treatment of H292 cells with C-PC enhanced NBR2 levels in a dose- and time-dependent manner. Downregulation of NBR2 in H292 cells inhibited the activity of C-PC on cell proliferation, viability and clone formation. Further mechanistic investigation showed that the down-regulation of NBR2 abolished the modulatory effects of C-PC on the AMPK/mTOR signalling pathway. In conclusion, C-PC inhibits H292 cell growth by enhancing the NBR2/AMPK signalling pathway.
- MeSH
- adenosinmonofosfát farmakologie terapeutické užití MeSH
- annexin A5 farmakologie terapeutické užití MeSH
- apoptóza MeSH
- fykokyanin metabolismus farmakologie terapeutické užití MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory plic * farmakoterapie genetika metabolismus MeSH
- nemalobuněčný karcinom plic * farmakoterapie genetika metabolismus MeSH
- pohyb buněk MeSH
- proliferace buněk MeSH
- proteinkinasy aktivované AMP genetika metabolismus farmakologie MeSH
- RNA dlouhá nekódující * genetika MeSH
- sinkalid metabolismus farmakologie terapeutické užití MeSH
- sirolimus farmakologie terapeutické užití MeSH
- TOR serin-threoninkinasy metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Clear cell renal cell carcinoma (ccRCC) is very common and accounts for most kidney cancer deaths. While many studies are being conducted in finding the prognostic signatures of ccRCC, we believe that ferroptosis, which involves programmed cell death dependent on iron accumulation, has therapeutic potential in ccRCC. Recent research has shown that long noncoding RNAs (lncRNAs) are involved in ferroptosis-related tumour processes and are closely related to survival in patients with ccRCC. Hence, in this study we aim to further explore the role of ferroptosis-related lncRNAs (FRLs) in ccRCC, hoping to establish a signature to predict the survival outcome of ccRCC. We analysed transcriptome data from The Cancer Genome Atlas database (TCGA) and ferroptosis-related genes (FRGs) from FerrDb to identify FRLs using Pearson's correlation. Lasso Cox regression analysis and multivariate Cox proportional hazards models screened seventeen optimal FRLs for developing prognostic signatures. Kaplan-Meier survival curves and ROC curves were then plotted for validating the sensitivity, specificity, and accuracy of the identified signatures. Gene Set Enrichment Analysis and CIBERSORT algorithm were deployed to explore the role of these FRLs in the tumour microenvironment. It was concluded that these models demonstrate excellent performance in predicting prognosis among patients with ccRCC, also indicating association with the clinicopathologic parameters such as tumour grade, tumour stage and tumour immune infiltration. In conclusion, our findings provide novel insights into ferroptosis-related lncRNAs in ccRCC, which are important targets for investigating the tumorigenesis of ccRCC.
- MeSH
- ferroptóza * genetika MeSH
- karcinom z renálních buněk * genetika patologie MeSH
- lidé MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádorové mikroprostředí MeSH
- nádory ledvin * genetika patologie MeSH
- RNA dlouhá nekódující * genetika MeSH
- železo MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Recently, more and more efforts are directed towards developing new imaging and drug-delivery options based on various nanoparticles, exploiting their unique properties. Here, ultra-small gold nanoparticles functionalized with widely used polyethylene glycol and its amine-terminated form were tested in respect of their potential interactions with human immune cells (cell line and primary cells). The results showed that differently terminated ultrasmall gold nanoparticles represent an interesting theranostic platform as they are harmless to immune cells (not inducing cytotoxicity and severe immune response) and on the other hand, they can serve as imaging and/or drug delivery agents using e.g. monocytes/ macrophages as "Trojan horses" to deliver these nanoparticles across the blood-brain barrier and diagnose or treat pathologies of the central nervous system.
- MeSH
- aminy MeSH
- kovové nanočástice * MeSH
- lidé MeSH
- polyethylenglykoly MeSH
- zlato * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
It is known that intracellular pathogens interact and react with the cellular immune system through exosomes produced by macrophages. This study aimed to determine whether co-culture of macrophages and Talaromyces marneffei induces exosomes and leads to immune responses. T. marneffei was incubated to collect conidia, co-cultured with human macrophages, which then induced exosomes. In cellular experiments, after extraction and purification, the exosomes were then observed by electron microscopy and detected by flow cytometry and mass spectrometry. In animal experiments, flow cytometry and enzyme-linked immunosorbent assay were used to examine whether exosomes were antigenpresenting. The results showed that purified exosomes produced a pro-inflammatory response and stimulated production of TNF-α in non-fungal-treated macrophages. Protein mass spectrometry analysis of exosomes also indicated their potential ability to activate the internal immune response system and the pro-inflammatory response. Translation and ribosomes were the most abundant GO terms in proteins, and the most relevant KEGG pathway was the biosynthesis of secondary metabolites. Furthermore, in vivo experiments revealed that exosomes induced activation of lymphocytes and increased expression of TNF-α and IL-12 in the lung, mediastinum, and spleen area. In conclusion, exosomes can be released by co-culture of T. marneffei and macrophages, having antigen-presenting functions, promoting macrophage inflammation, and initiating adaptive immune responses. These processes are inextricably linked to the translation of secondary metabolites, ribosomes and biosynthesis.
As the number of cancer patients globally increases, a need for reliable biomarkers including circulating tumour DNA from liquid biopsy for diagnosis, prognosis and monitoring of the disease is rising. Currently, mainly tissue samples from biopsy are used, but there are certain limitations: firstly, it is an invasive technique, and secondly, in some cases it is almost impossible to obtain an acceptable tissue sample. This could be changed by using circulating cell-free DNA from liquid biopsy, which also gives the possibility of repeated examination. Here, we focus on the options of isolating circulating cell-free DNA from plasma samples using two isolation techniques: precision manual QIAamp Circulating Nucleic Acid Kit and automatic MagNA Pure Compact (MPC) using Nucleic Acid Isolation Kit I. Manual extraction gave significantly better yields of circulating tumour DNA (P < 0.05). This DNA also had less contaminants (organic compounds or proteins). DNA obtained by both tested methods of isolation is suitable for subsequent molecular genetic methods.
- MeSH
- cirkulující nádorová DNA * MeSH
- lidé MeSH
- nádory * MeSH
- tekutá biopsie MeSH
- volné cirkulující nukleové kyseliny * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Diarrhoea is a common clinical condition; its pathogenesis is strongly associated with gut microbiota dysbiosis. Limonitum is a well-known traditional Chinese medicine that exerts appreciable benefits regarding the amelioration of diarrhoea. However, the mechanism through which Limonitum ameliorates diarrhoea remains unclear. Here, the efficacy and underlying mechanism of Limonitum decoction (LD) regarding diarrhoea were explored from the aspect of gut microbiota. Castor oil (CO) was used to induce diarrhoea in mice, which were then used to evaluate the effects of LD regarding the timing of the first defecation, diarrhoea stool rate, degree of diarrhoea, diarrhoea score, intestinal propulsive rate, and weight of intestinal contents. The concentrations of short-chain fatty acids (SCFAs), including acetic, propionic, isobutyric, butyric and valeric acids, were analysed by gas chromatography-mass spectrometry (GC-MS). The 16S rRNA high-throughput sequencing technology was applied to evaluate changes in the gut microbiota under exposure to LD. LD was found to effectively ameliorate the symptoms of diarrhoea, and the diversity and relative abundance of gut microbiota were restored to normal levels following LD treatment. Additionally, LD significantly restored the observed reductions in SCFAs. These results provide strong evidence that LD can sufficiently ameliorate diarrhoea in mice by regulating their gut microbiota. The findings presented here highlight that Limonitum may constitute a prospective remedy for diarrhoea.
- MeSH
- myši MeSH
- prospektivní studie MeSH
- průjem MeSH
- ricinový olej MeSH
- RNA ribozomální 16S MeSH
- střevní mikroflóra * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Transient receptor potential cation channel subfamily V member 1 (TRPV1) has been found over-expressed in low back pain (LBP) patients with neuropathic pain (NP), but the underlying mechanism is still unclear. In the present study, the up-regulation of the TRPV1 protein level in sinuvertebral nerve biopsies from patients with NP was verified by immunoblotting, but the TRPV1 mRNA level was not significantly changed. MiRNAs targeting TRPV1 mRNA were predicted by a bioinformatic tool, and the interactions between the miRNAs and TRPV1 were confirmed by dual luciferase assay. The correlation between NFKB1 signalling and TRPV1 expression was analysed and confirmed by using sNF96.2 cells after lipopolysaccharide stimulation. We found that five out of 18 miRNAs repressed TRPV1 expression, and the levels of miR-375 and miR-455 were negatively correlated with the protein level of TRPV1 in patients with NP. MiR-375 and miR-455 were identified to repress TRPV1 expression via targeting the 3'UTR of TRPV1 mRNA. NFKB1 signalling activation down-regulated the expression of miR-375 and miR-455, and thus up-regulated the TRPV1 protein level. In conclusion, we partially unveiled the mechanism of how TRPV1 is over-expressed in chronic LBP patients with NP and provided two potential candidate miRNAs for NP treatment.
