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1.
Braz. j. med. biol. res ; 54(10): e11156, 2021. graf
Article in English | LILACS | ID: biblio-1285646

ABSTRACT

The objective of this study was to investigate the effect of human esophageal fibroblast-derived exosomal miR-21 on cisplatin sensitivity against esophageal squamous EC9706 cells. EC9706 cells were co-cultured indirectly with human esophageal fibroblasts (HEF) or miR-21 mimics transfected-HEF in the transwell system. The exosomes in HEF-culture conditioned medium were extracted by differential ultracentrifugation. EC9706 cells were co-cultured with HEF-derived exosomes directly. The cisplatin sensitivity against EC9706 cells was revealed via half maximal inhibitory concentration (IC50) values using MTT assay. The expressions of miR-21, programmed cell death 4 (PDCD4) mRNA, and gene of phosphate and tension homology deleted on chromosome ten (PTEN) mRNA were determined by qRT-PCR. The changes of the protein level were detected using western blot assay. IC50 values of cisplatin against EC9706 cells were increased after EC9706 cells were co-cultured with either HEF or exosomes derived from miR-21 mimics-transfected HEF. Following the increased level of miR-21, the mRNA expression and protein levels of PTEN and PDCD4 were decreased in EC9706 cells. The cisplatin sensitivity to EC9706 cells was reduced by HEF-derived exosomal miR-21 through targeting PTEN and PDCD4. This study suggested that non-tumor cells in the tumor micro-environment increased the tumor anti-chemotherapy effects through their exosomes.


Subject(s)
Humans , Esophageal Neoplasms/genetics , Esophageal Neoplasms/drug therapy , Carcinoma , MicroRNAs/genetics , Cisplatin/pharmacology , RNA-Binding Proteins , Apoptosis , Cell Line, Tumor , Cell Proliferation , Apoptosis Regulatory Proteins/metabolism , Tumor Microenvironment , Fibroblasts/metabolism
2.
Article in Chinese | WPRIM | ID: wpr-880170

ABSTRACT

OBJECTIVE@#To analyze the dynamic molecular expression characteristics of single cell RNA binding proteins (RBPs) in the development of mouse embryonic hematopoitic stem cells (HSCs), and obtain the functional research target RNA splicing factor--Mbnl1, to clarify the function of Mbnl1 involved in regulating mouse embryonic HSC development.@*METHODS@#Bioinformatics was used to analyze the single-cell transcriptome data of mouse embryos during HSC development, and the single-cell RBP dynamic molecular expression maps in HSC development was obtained. Mbnl1 was obtained by combining differential analysis and literature research screening. The Mbnl1-knockout mouse model was constructed by the CRISPER/Cas9 technology. Aorta-gonad-mesonephros (AGM) and yolk sac (YS) tissue in two genotype embryos of Mbnl1@*RESULTS@#The in vitro CFU-C experiment of hematopoietic cells preliminarily indicated that there was no significant difference in the number of cell colonies in AGM region and YS transformed by the two genotypes of Mbnl1@*CONCLUSION@#Through functional experiments in vivo and in vitro, it has been confirmed that knockout of the RNA splicing factor--Mbnl1 does not affect the development of HSPC in AGM region of mouse embryo.


Subject(s)
Animals , DNA-Binding Proteins , Gonads , Hematopoiesis , Hematopoietic Stem Cells , Mesonephros , Mice , RNA-Binding Proteins/genetics , Yolk Sac
3.
Article in Chinese | WPRIM | ID: wpr-880136

ABSTRACT

OBJECTIVE@#To investigate the regulatory effects of RBM47 on HMGA2 and the function of RBM47 in human chronic myeloid leukemia cell K562.@*METHODS@#K562 cells were transduction by the overexpressed and knockdown RBM47 lentiviral vector. CCK-8 assay was used to detect the effect of RBM47 on the proliferation of K562 cells. Flow cytometry assay was used to detect the effect of RBM47 on the cell cycle progression of K562 cells. RNA immunoprecipitation assay was used to detect the association between RBM47 and HMGA2 mRNA. RT-qPCR was used to detect the effects of RBM47 on the stability of HMGA2 mRNA. Western blot was used to evaluate the effect of RBM47 on HMGA2 protein expression.@*RESULTS@#The overexpressed RBM47 could inhibit the proliferation and cell cycle progression of K562 cells. However, the inhibitation of RBM47 could improve the proliferation and cell cycle progression of K562 cells. RBM47 combined with HMGA2 mRNA could promote the degradation of HMGA2 mRNA. Thus, the overexpressed RBM47 could decrease the expression of HMGA2 protein in K562 cells.@*CONCLUSION@#RNA binding protein RBM47 can inhibit the proliferation of K562 cells by regulating HMGA2 expression.


Subject(s)
Apoptosis , Cell Proliferation , HMGA2 Protein/genetics , Humans , K562 Cells , RNA, Messenger/genetics , RNA-Binding Proteins/genetics
4.
Electron. j. biotechnol ; 46: 8-13, jul. 2020. tab, graf
Article in English | LILACS | ID: biblio-1223212

ABSTRACT

BACKGROUND: Poly-3-hydroxybutyrate (PHB) can be efficiently produced in recombinant Escherichia coli by the overexpression of an operon (NphaCAB) encoding PHB synthetase. Strain improvement is considered to be one of critical factors to lower the production cost of PHB in recombinant system. In this study, one of key regulators that affect the cell growth and PHB content was confirmed and analyzed. RESULT: S17-3, a mutant E. coli strain derived from S17-1, was found to be able to achieve high cell density when expressing NphaCAB with the plasmid pBhya-CAB. Whole genome sequencing of S17-3 revealed genetic alternations on the upstream regions of csrA, encoding a global regulator cross-talking between stress response, catabolite repression and other metabolic activities. Deletion of csrA or expression of mutant csrA resulted in improved cell density and PHB content. CONCLUSION: The impact of gene deletion of csrA was determined, dysfunction of the regulators improved the cell density of recombinant E. coli and PHB production, however, the detail mechanism needs to be further clarified.


Subject(s)
Escherichia coli/metabolism , Hydroxybutyrates/metabolism , Repressor Proteins/genetics , Biopolymers/genetics , Recombinant Proteins , RNA-Binding Proteins/genetics , Gene Deletion , Escherichia coli Proteins/genetics , Escherichia coli/genetics , Metabolic Engineering , Ligases/metabolism
5.
Biol. Res ; 53: 42, 2020. tab, graf
Article in English | LILACS | ID: biblio-1131886

ABSTRACT

BACKGROUND: Basal-like breast cancer (BLBC) or triple-negative breast cancer (TNBC) is an aggressive and highly metastatic subtype of human breast cancer. The present study aimed to elucidate the potential tumor-suppressive function of MATR3, an abundant nuclear protein, in BLBC/TNBC, whose cancer-relevance has not been characterized. METHODS: We analyzed in vitro tumorigenecity by cell proliferation and soft agar colony formation assays, apoptotic cell death by flow cytometry and Poly (ADP-ribose) polymerase (PARP) cleavage, epithelial-mesenchymal transition (EMT) by checking specific EMT markers with real-time quantitative PCR and in vitro migration and invasion by Boyden Chamber assays. To elucidate the underlying mechanism by which MATR3 functions as a tumor suppressor, we performed Tandem affinity purification followed by mass spectrometry (TAP-MS) and pathway analysis. We also scrutinized MATR3 expression levels in the different subtypes of human breast cancer and the correlation between MATR3 expression and patient survival by bioinformatic analyses of publicly available transcriptome datasets. RESULTS: MATR3 suppressed in vitro tumorigenecity, promoted apoptotic cell death and inhibited EMT, migration, and invasion in BLBC/TNBC cells. Various proteins regulating apoptosis were identified as MATR3-binding proteins, and YAP/TAZ pathway was suppressed by MATR3. MATR3 expression was inversely correlated with the aggressive and metastatic nature of breast cancer. Moreover, high expression levels of MATR3 were associated with a good prognosis of breast cancer patients. CONCLUSIONS: Our data demonstrate that MATR3 functions as a putative tumor suppressor in BLBC/TNBC cells. Also, MATR3 potentially plays a role as a biomarker in predicting chemotherapy-sensitivity and patient survival in breast cancer patients.


Subject(s)
Humans , Female , Genes, Tumor Suppressor , RNA-Binding Proteins/genetics , Nuclear Matrix-Associated Proteins/genetics , Triple Negative Breast Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Cell Movement , Apoptosis , Cell Line, Tumor , Cell Proliferation , Epithelial-Mesenchymal Transition
6.
Biol. Res ; 53: 43, 2020. tab, graf
Article in English | LILACS | ID: biblio-1131887

ABSTRACT

BACKGROUND: Breast cancer, the most common cancer in women worldwide, causes the vast majority of cancer-related deaths. Undoubtedly, tumor metastasis and recurrence are responsible for more than 90 percent of these deaths. MicroRNAs are endogenous noncoding RNAs that have been integrated into almost all the physiological and pathological processes, including metastasis. In the present study, the role of miR-128 in breast cancer was investigated. RESULTS: Compared to the corresponding adjacent normal tissue, the expression of miR-128 was significantly suppressed in human breast cancer specimens. More importantly, its expression level was reversely correlated to histological grade of the cancer. Ectopic expression of miR-128 in the aggressive breast cancer cell line MDA-MB-231 could inhibit cell motility and invasive capacity remarkably. Afterwards, Metadherin (MTDH), also known as AEG-1 (Astrocyte Elevated Gene 1) and Lyric that implicated in various aspects of cancer progression and metastasis, was further identified as a direct target gene of miR-128 and its expression level was up-regulated in clinical samples as expected. Moreover, knockdown of MTDH in MDA-MB-231 cells obviously impaired the migration and invasion capabilities, whereas re-expression of MTDH abrogated the suppressive effect caused by miR-128. CONCLUSIONS: Overall, these findings demonstrate that miR-128 could serve as a novel biomarker for breast cancer metastasis and a potent target for treatment in the future.


Subject(s)
Humans , Female , Breast Neoplasms/genetics , MicroRNAs/physiology , MicroRNAs/genetics , Neoplasm Invasiveness/genetics , Cell Adhesion Molecules/genetics , Cell Adhesion Molecules/metabolism , Gene Expression Regulation, Neoplastic , RNA-Binding Proteins , Cell Line, Tumor , Membrane Proteins , Neoplasm Recurrence, Local
7.
Braz. j. med. biol. res ; 53(4): e9290, 2020. tab, graf
Article in English | LILACS | ID: biblio-1089356

ABSTRACT

This study was designed to investigate the expression of RBM8A protein in patients with gastric cancer (GC) and to explore its correlation with clinical pathological features as well as prognosis. One hundred pairs of gastric carcinoma tissues and adjacent tissues from patients undergoing gastrectomy for GC were included in this study. The protein expression level of RBM8A was determined by immunohistochemistry using tissue microarrays. We also detected the mRNA expression level of RBM8A in 16 pairs of gastric carcinoma tissues and adjacent tissues. Meanwhile, we predicted the potential correlation between RBM8A and tumor stages as well as survival condition in patents with GC based on The Cancer Genome Atlas (TCGA) database. The correlation of RBM8A with the clinical pathological features and prognosis of the 100 patients with GC was also elucidated. The expression level of RBM8A was significantly higher in gastric carcinoma tissues compared to the adjacent tissues. The protein level of RBM8A was correlated with tumor size (P=0.031), depth of invasion (P<0.001), lymph node metastasis (P<0.001), TNM stage (<0.001), and distant metastasis (P=0.001). Patients with increased RBM8A expression (P<0.0018, 95%CI=0.322−0.871), higher TNM stage (P<0.001, 95%CI=4.990−11.283), and lymph node metastasis (P<0.001, 95%CI=2.873−4.002) had a lower overall survival. Taken together, our study demonstrated that RBM8A may act as a proto-oncogene, which could be a promising biomarker and therapeutic target in the diagnosis and treatment of GC.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Stomach Neoplasms/metabolism , RNA-Binding Proteins/metabolism , Prognosis , Stomach Neoplasms/genetics , Stomach Neoplasms/pathology , RNA, Messenger/metabolism , Immunohistochemistry , Biomarkers, Tumor/genetics , Gene Expression Regulation, Neoplastic , Survival Analysis , Proto-Oncogene Proteins/metabolism , RNA-Binding Proteins/genetics , Gastric Mucosa/pathology , Lymphatic Metastasis/pathology , Neoplasm Metastasis , Neoplasm Staging
8.
Journal of Experimental Hematology ; (6): 1137-1143, 2020.
Article in Chinese | WPRIM | ID: wpr-827150

ABSTRACT

OBJECTIVE@#To investigate the effects of CPEB4 on the migration and cycle of K562 cells and the changes of protein molecules that may be involved in the regulatory mechanism.@*METHODS@#Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells. The overexpression plasmid pcDNA3.1(+)-His-CPEB4, silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells by electroporation so as to change CPEB4. The transfection efficiency was detected by Western blot. Finally, the migration and cycle of different cells were detected by Transwell chamber and flow cytometry.Western blot was used to detect the expression changes of MMP2, MMP9, CDK4, CyclinD1 and P21 proteins.@*RESULTS@#Compared with normal white blood cells, the expression of CPEB4 protein in K562 cells was significantly enhanced (P<0.01); Compared with the control group, CPEB4-silenced K562 cells showed that the cell migration ability was significantly enhanced (P<0.01); G/G phase cell ratio reduced, G/M phase cell ratio increased, and cell cycle progression accelerated(P<0.01), The expression levels of MMP2 (P<0.05), MMP9 (P<0.05), CDK4 (P<0.01), CyclinD1 (P<0.01) proteins increased significantly. The expression level of P21 protein significantly decreased (P<0.01). The migration ability of K562 cells after CPEB4 overexpression was decreased (P<0.01), the cell ratio of G/G phase in the cell cycle increased, the cell proportion of S phase decreased and the cell cycle progression was arrested at G/G phase (P<0.01). The expression of P21 protein increased, MMP2 , MMP9, CDK4, CyclinD1 protein expression decreased significantly(P<0.05-0.01).@*CONCLUSION@#CPEB4 can inhibit the migration of K562 cells and arrest cell cycle progression at G/G phase. Its mechanism may be related with regulating the exprossion of MMP2, MMP9, CDK4, CyclinD1 and P21 proteins.


Subject(s)
Apoptosis , Cell Cycle , Cell Proliferation , Humans , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , RNA-Binding Proteins
9.
Acta Physiologica Sinica ; (6): 325-335, 2020.
Article in English | WPRIM | ID: wpr-827055

ABSTRACT

The mammalian intestinal epithelium is a rapidly self-renewing tissue in the body and directly interfaces with a wide array of luminal noxious contents and microorganisms. Homeostasis of the intestinal epithelium is preserved through well-controlled mechanisms including posttranscriptional regulation. RNA-binding protein (RBP) HuR regulates the stability and translation of target mRNAs and is intimately involved in many aspects of gut mucosal pathophysiology. Here we highlight the biological roles of HuR in maintaining the integrity of the intestinal epithelium, with particular focus on the emerging evidence of HuR in the regulation of intestinal epithelial renewal, mucosal repair, defense, and gut permeability. We also further analyze the mechanisms through which HuR and its interactions with other RBPs and noncoding RNAs (ncRNAs) such as microRNAs and long ncRNAs modulate the intestinal epithelial homeostasis. With rapidly advancing knowledge of RBPs and ncRNAs, there is growing recognition that posttranscriptional control of the intestinal epithelium homeostasis might be promising therapeutic targets in our efforts to protect the integrity of the intestinal epithelium under critical pathological conditions.


Subject(s)
Animals , Gene Expression Regulation , Homeostasis , Intestinal Mucosa , RNA, Long Noncoding , RNA-Binding Proteins
10.
Article in Chinese | WPRIM | ID: wpr-879473

ABSTRACT

OBJECTIVE@#To detect variants of ADAR1 gene in two Chinese pedigrees affected with dyschromatosis symmetrica hereditaria (DSH).@*METHODS@#Clinical data and peripheral blood samples of the pedigrees were collected. All exons of the ADAR1 gene were amplified by PCR and subjected to Sanger sequencing. Suspected pathogenic variants were validated among other members of the pedigrees and 100 unrelated healthy controls.@*RESULTS@#For pedigree 1, Sanger sequencing has identified a heterozygous missense variant c.3002G>C (p.Asp968His) in exon 11 of the ADAR1 gene in the proband and his father. For pedigree 2, a novel nonsense variant c.3145C>T (p.Gln1049Ter) was identified in exon 12 of the ADAR1 gene in the proband and his son, which were previously unreported and absent among the healthy controls.@*CONCLUSION@#The c.3002G>C (p.Asp968His) and c.3145C>T (p.Gln1049Ter)variants of the ADAR1 gene probably underlay the DSH in the two pedigrees.


Subject(s)
Adenosine Deaminase/genetics , Humans , Mutation , Pedigree , Pigmentation Disorders/genetics , RNA-Binding Proteins/genetics
11.
Electron. j. biotechnol ; 39: 74-81, may. 2019. tab, ilus, graf
Article in English | LILACS | ID: biblio-1052041

ABSTRACT

Background: CPEB is considered as an RNA-binding protein first identified in Xenopus oocytes. Although CPEB1 was involved in the growth of oocyte, its role in goat follicular granulosa cell has not been fully elucidated. To clarify the functions of this gene in goat follicular granulosa cells, CPEB1-overexpressing vector and interference vector were structured and transfected into follicular granulosa cells from Jiangsu native white goats of Nantong city, Jiangsu Province, China. The expression levels of differentiation-related genes including CDK1, Cyclin B1, and C-mos were determined 24 h after administration of CPEB1 by quantitative real-time polymerase chain reaction and Western blotting methods. Results: The expression levels of CDK1, Cyclin B1, and C-mos were significantly upregulated after overexpression and significantly downregulated after interference with CPEB1. Conclusions: The CPEB1 gene expression could affect the transcription of genes related to early cleavage divisions, which provided a reference for further research on its role in the growth and maturation of oocytes.


Subject(s)
Animals , Female , Oocytes , Transcription Factors/genetics , Goats/genetics , Transfection , Fertilization in Vitro , Gene Expression , Blotting, Western , Polymerase Chain Reaction/methods , RNA-Binding Proteins , Embryo Transfer , Livestock , Fluorescence , Granulosa Cells
12.
Journal of Experimental Hematology ; (6): 1779-1785, 2019.
Article in Chinese | WPRIM | ID: wpr-781397

ABSTRACT

OBJECTIVE@#To explore the expression of CPEB4 in K562 cells, the biological activity and its possible molecular mechanisms.@*METHODS@#Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells. The overexpression plasmid pcDNA3.1(+)-His-CPEB4 and the silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells to change the expression of CPEB4 in K562 cells, and the transfection efficiency was detected by Western blot. Finally, CCK-8 and flow cytometry were used to detect the proliferation and apoptosis of differently treated cells, and the expression changes of proliferation and apoptosis marker proteins (AKT, p-AKT, caspase-3, BCL-2) were detected by Western blot.@*RESULTS@#Compared with normal leukocytes, the expression of CPEB4 protein in K562 cells was higher (P<0.01). Compared with the control group, the proliferation of CPEB4-silenced K562 cells significantly increased (P<0.01), the number of apoptotic cell significantly decreased, and expression of AKT, p-AKT and BCL-2 was significantly increased, the protein expression of caspase-3 was significantly reduced. The proliferation of K562 cells after CPEB4 overexpression was slowed down (P<0.05), the number of apoptotic cells significantly increased,the expressions of AKT, p-AKT and BCL-2 were significantly down-regulated, and the expression of caspase-3 was up-regulated.@*CONCLUSION@#CPEB4 can inhibit the proliferation and promote the apoptosis of K562 cells, the AKT, p-AKT, BCL-2 and caspase-3 are involved in the regulation mechanism.


Subject(s)
Apoptosis , Cell Proliferation , Humans , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , RNA-Binding Proteins , Metabolism , Transfection
13.
Article in Chinese | WPRIM | ID: wpr-781260

ABSTRACT

OBJECTIVE@#To investigate the regulatory role of Musashi-1 (MSI1) in the proliferation and growth of hepatocellular carcinoma (HCC) cells.@*METHODS@#We examined the expression of MSI1 in HCC and paired adjacent tissues from 24 patients using immunohistochemistry and Western blotting. A MSI1-expressing vector was constructed and stably transfected into HepG2 cells, and short hairpin RNAs (shRNAs) that targeted MSI1 mRNA were ligated into the vector and stably transfected in Huh7 cells. The effects of MSI1 overexpression and silencing on the proliferation, viability and cell cycle of HepG2 cells were investigated using flow cytometry or MTT assay. The expressions of PCNA, cyclin D1, APC and β-catenin in the HCC cells were detected with Western blotting.@*RESULTS@#MSI1 expression was significantly up-regulated in HCC tissues as compared with that in the adjacent tissues. Overexpression of MSI1 in HepG2 cells resulted in significantly enhanced cell growth ( < 0.01) and significantly reduced G0/G1 phase cells from (58.42±3.18)% to (40.67±1.22)% and increased S phase cells from (28.51± 1.93)% to (40.06±1.92)% ( < 0.01), causing also increases in the expressions of PCNA and Cyclin D1. Knockdown of MSI1 in Huh7 cells obviously inhibited the cell growth and caused cell cycle arrest at the G1/S phase ( < 0.01) with reduced protein expressions of PCNA and cyclin D1. Overexpression of MSI1 in HepG2 cells also down-regulated the expression of APC and up-regulated the expression of β-catenin protein, while MSI1 knockdown caused reverse changes in Huh7 cells.@*CONCLUSIONS@#MSI1 promotes the progression of HCC through positive modulation of cell growth and cell cycle the Wnt/β-catenin pathway.


Subject(s)
Carcinoma, Hepatocellular , Cell Cycle , Cell Proliferation , Hep G2 Cells , Humans , Liver Neoplasms , Nerve Tissue Proteins , Metabolism , RNA-Binding Proteins , Metabolism
14.
Article in English | WPRIM | ID: wpr-739534

ABSTRACT

5-HT₆ receptor (5-HT₆R) is implicated in cognitive dysfunction, mood disorder, psychosis, and eating disorders. However, despite its significant role in regulating the brain functions, regulation of 5-HT₆R at the molecular level is poorly understood. Here, using yeast two-hybrid assay, we found that human 5-HT₆R directly binds to neuro-oncological ventral antigen 1 (Nova-1), a brain-enriched splicing regulator. The interaction between 5-HT₆R and Nova-1 was confirmed using GST pull-down and co-immunoprecipitation assays in cell lines and rat brain. The splicing activity of Nova-1 was decreased upon overexpression of 5-HT₆R, which was examined by detecting the spliced intermediates of gonadotropin-releasing hormone (GnRH), a known pre-mRNA target of Nova-1, using RT-PCR. In addition, overexpression of 5-HT₆R induced the translocation of Nova-1 from the nucleus to cytoplasm, resulting in the reduced splicing activity of Nova-1. In contrast, overexpression of Nova-1 reduced the activity and the total protein levels of 5-HT₆R. Taken together, these results indicate that when the expression levels of 5-HT₆R or Nova-1 protein are not properly regulated, it may also deteriorate the function of the other.


Subject(s)
Animals , Brain , Cell Line , Cytoplasm , Eating , Gonadotropin-Releasing Hormone , Humans , Immunoprecipitation , Mood Disorders , Psychotic Disorders , Rats , RNA Precursors , RNA-Binding Proteins , Serotonin , Two-Hybrid System Techniques
15.
Annals of Dermatology ; : 387-392, 2019.
Article in English | WPRIM | ID: wpr-762360

ABSTRACT

BACKGROUND: Alopecia areata (AA), a chronic, relapsing hair-loss disorder, is considered to be a T-cell-mediated autoimmune disease. Cold-inducible RNA-binding protein (CIRP) belongs to a family of cold-shock proteins that respond to cold stress, and has been identified as a damage-associated molecular pattern (DAMP) molecule that triggers the inflammatory response. Recent studies have shown that high-mobility group box 1, another DAMP molecule, is elevated in serum and scalp tissue of AA patients, suggesting a relationship between DAMP molecules and the pathogenesis of AA. OBJECTIVE: To investigate the clinical significance of serum CIRP levels in AA. METHODS: The serum levels of CIRP were compared between 68 patients with AA and 20 healthy controls. Additionally, the correlation between CIRP level and various clinical parameters was evaluated. RESULTS: The serum CIRP levels were significantly higher in AA patients compared to healthy subjects. Moreover, there was an association between the serum CIRP level and clinical characteristics, such as disease duration and disease activity. However, there was no significant difference in the serum CIRP level among the clinical types of AA (AA multiplex, alopecia totalis, and alopecia universalis). CONCLUSION: These results suggest that CIRP may play a significant role in the pathogenesis of AA and could be a potential biologic marker for monitoring the disease activity of AA.


Subject(s)
Alopecia Areata , Alopecia , Autoimmune Diseases , Biomarkers , Healthy Volunteers , Humans , Inflammation , RNA-Binding Proteins , Scalp
16.
Article in English | WPRIM | ID: wpr-761785

ABSTRACT

Curcumin, an active ingredient of Curcuma longa L., can reduce the concentration of low-density lipoproteins in plasma, in different ways. We had first reported that curcumin exhibits hypocholesterolemic properties by improving the apolipoprotein B (apoB) mRNA editing in primary rat hepatocytes. However, the role of curcumin in the regulation of apoB mRNA editing is not clear. Thus, we investigated the effect of curcumin on the expression of multiple editing components of apoB mRNA cytidine deamination to uridine (C-to-U) editosome. Our results demonstrated that treatment with 50 µM curcumin markedly increased the amount of edited apoB mRNA in primary mouse hepatocytes from 5.13%–8.05% to 27.63%–35.61%, and significantly elevated the levels of the core components apoB editing catalytic polypeptide-1 (APOBEC-1), apobec-1 complementation factor (ACF), and RNA-binding-motif-protein-47 (RBM47), as well as suppressed the level of the inhibitory component glycine-arginine-tyrosine-rich RNA binding protein. Moreover, the increased apoB RNA editing by 50 µM curcumin was significantly reduced by siRNA-mediated APOBEC-1, ACF, and RBM47 knockdown. These findings suggest that curcumin modulates apoB mRNA editing by coordinating the multiple editing components of the editosome in primary hepatocytes. Our data provided evidence for curcumin to be used therapeutically to prevent atherosclerosis.


Subject(s)
Animals , Apolipoproteins B , Apolipoproteins , Atherosclerosis , Complement System Proteins , Curcuma , Curcumin , Cytidine , Deamination , Hepatocytes , Lipoproteins, LDL , Mice , Plasma , Rats , RNA Editing , RNA, Messenger , RNA-Binding Proteins , Uridine
17.
Article in Chinese | WPRIM | ID: wpr-772062

ABSTRACT

OBJECTIVE@#To explore the role of long non-coding RNA (lncRNA) X inactive specific transcript (XIST) in modulating cisplatin (DDP) resistance of human nasopharyngeal carcinoma cells and investigate the possible mechanism.@*METHODS@#Realtime PCR was performed to detect the expression of XIST in cisplatin-resistant human nasopharyngeal carcinoma cell line HNE1/DDP. The effects of up-regulation and down-regulation of XIST on DDP resistance, proliferation and apoptosis of HNE1/ DDP cells were assessed using MTT assay, EdU assay and flow cytometry. Western blotting was used to detect the changes in the expressions of programmed cell death 4 (PDCD4) and Fas ligand (Fas-L) proteins in the cells in response to up-regulation or down-regulation of XIST.@*RESULTS@#The expression of XIST was significantly up-regulated in HNE1/DDP cells in comparison with HNE1 cells (0.57±0.06 0.1±0.02, < 0.05). Down-regulation of XIST significantly decreased while up-regulation of XIST obviously increased DDP resistance of HNE1/DDP cells ( < 0.05). Down-regulation of XIST significantly reduced the proliferation (6.17 ± 1.93 16.59 ± 4.86, < 0.05) and enhanced apoptosis [(18.04 ± 4.72)% (4.22 ± 1.65)%, < 0.05], while upregulating XIST enhanced the proliferation (25.40±7.21 13.16±3.95, < 0.05) and inhibited apoptosis [(2.82±0.88)% (6.46± 1.75)%, < 0.05] in HNE1/DDP cells. Down-regulation of XIST significantly increased the protein expressions of PDCD4 and Fas-L ( < 0.05) in HNE1/DDP cells, and up-regulation of XIST resulted in reverse changes in PDCD4 and Fas-L expressions ( < 0.05).@*CONCLUSIONS@#XIST is up-regulated in HNE1/DDP cells, and down-regulation and up-regulation of XIST expression reduce and increase DDP resistance of the cells, respectively, possibly as a result of changes in the expressions of PDCD4 and Fas-L.


Subject(s)
Antineoplastic Agents , Apoptosis , Apoptosis Regulatory Proteins , Cell Line, Tumor , Cell Proliferation , Cisplatin , Drug Resistance, Neoplasm , Humans , Nasopharyngeal Carcinoma , RNA, Long Noncoding , RNA-Binding Proteins , fas Receptor
18.
Article in Chinese | WPRIM | ID: wpr-771966

ABSTRACT

OBJECTIVE@#To detect mutations of ADAR gene in two pedigrees affected with dyschromatosis symmetrica hereditaria (DSH).@*METHODS@#Potential mutations of the ADAR gene were analyzed by Sanger sequencing of the probands from both pedigrees. Suspected mutations were validated by Sanger sequencing of other patients from both pedigrees as well as unrelated healthy individuals.@*RESULTS@#A heterozygous nonsense mutation c.1325C>G (p.Ser442Ter) and a novel nonsense mutation c.1498C>T (p.Gln500Ter) were respectively identified in the ADAR gene among all patients from the two pedigrees but not among 200 healthy individuals.@*CONCLUSION@#Mutations of the ADAR gene probably underlie the DSH in the two pedigrees. Above findings have enriched the spectrum of ADAR gene mutation.


Subject(s)
Adenosine Deaminase , Humans , Mutation , Pedigree , Pigmentation Disorders , Genetics , RNA-Binding Proteins
19.
Journal of Experimental Hematology ; (6): 1064-1070, 2019.
Article in Chinese | WPRIM | ID: wpr-771838

ABSTRACT

OBJECTIVE@#To construct a K562 and adriamycin-resistant K562 (KAR) cell line with stably down-regulation of NCL (nucleolin) expression, and to investigate the effect of NCL down-regulation on the drug resistance in K562 and KAR cells.@*METHODS@#K562 and KAR cells were infected with lentivirus, and stably transfected cell clones were obtained by puromycin screening. The cell proliferation was detected by MTS assay, the cell apoptosis was detected by flow cytometry, and the expression level of drug resistance related genes was detected by real-time PCR.@*RESULTS@#The K562 and KAR cells with stable down-regulation of NCL were successfully constructed. Compared with the control group, the proliferation of K562 and KAR cells with down-regulating NCL expression decreased significantly (P <0.05), the apoptosis of cells increased significantly (P <0.05), and cell resistance to adriamycin was down-regulated.@*CONCLUSION@#Inhibition of NCL expression may increase the sensitivity of cells to adriamycin, which may be related with the promotion of apoptosis of K562 and KAR cells.


Subject(s)
Apoptosis , Doxorubicin , Drug Resistance, Neoplasm , Humans , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Phosphoproteins , RNA-Binding Proteins
20.
Article in Chinese | WPRIM | ID: wpr-773221

ABSTRACT

To study the mechanism and action of Cinnamomi Ramulus in ameliorating intrahepatic cholestasis induced by α-isothiocyanate( ANIT) in rats by regulating FXR pathway. Forty SD rats were randomly divided into normal group,model group,positive control( ursodeoxycholic acid) group( 60 mg·kg~(-1)),Cinnamomi Ramulus treatment( 60 mg·kg~(-1)·d~(-1)) group,and Cinnamomi Ramulus treatment( 20 mg·kg~(-1)·d~(-1)) group,with 8 rats in each group. Except for the normal control group,the other groups were intragastrically administered with the corresponding concentrations of continuous aqueous solution( 0. 005 m L·g~(-1)),once a day,for 7 days.Except for the normal group,the other groups were treated with ANIT( 100 mg·kg~(-1)),once a day,for 3 days. Blood was taken from the abdominal aorta 24 hours after the last administration,and serum alanine aminotransferase( ALT),aspartate aminotransferase( AST),total bilirubin( TBi L),and total bile acid( TBA) were measured. 1. 5-2 cm of rat liver tissue was taken. After fixation with10% formaldehyde,paraffin-embedded sections were taken,HE staining was performed,and immunohistochemistry( IHC) was used to analyze the expression of FXR. RNA and protein were extracted from rat liver tissue to detect FXR mRNA expression,as well as bile acid synthesis and detoxification,transport related SHP,UGT2 B4,BSEP protein expressions at downstream of FXR. Compared with the normal group,serum ALT,AST,TBi L,and TBA levels were elevated in the model group( P<0. 01),liver damage was severe,FXR protein's optical density decreased,FXR mRNA expression decreased,and SHP,UGT2 B4,BSEP protein expressions were decreased( P<0. 05,P<0. 01). Compared with the model group,the drug group could reduce serum ALT,AST,TB,TBA levels to different degrees( P<0. 05,P<0. 01),alleviate liver tissue damage,increase the optical density of FXR protein,and promote the expressions of FXR mRNA and FXR,SHP,BSEP and UGT2 B4 proteins( P<0. 05,P<0. 01). Cinnamomi Ramulus can alleviate ANIT-induced intrahepatic cholestasis,and reduce hepatocyte injury and serum ALT,AST,TBi L and TBA levels. The mechanism may be through FXR-SHP,FXR-UGT2 B4,FXR-BSEP signaling pathways. Therefore,in the pathogenesis of intrahepatic cholestasis,we can try to further explore in alleviating intrahepatic cholestasis with Cinnamomi Ramulus,so as to provide effective drugs for clinical treatment of intrahepatic cholestasis.


Subject(s)
Alanine Transaminase , Blood , Animals , Aspartate Aminotransferases , Blood , Bile Acids and Salts , Blood , Bilirubin , Blood , Cholestasis, Intrahepatic , Drug Therapy , Cinnamomum , Chemistry , Isothiocyanates , Liver , Plant Extracts , Pharmacology , RNA-Binding Proteins , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley
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