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1.
Article in Chinese | WPRIM | ID: wpr-927900

ABSTRACT

Objective: To investigate the mechanism that hypoxia promotes the migration of lung adenocarcinoma A549 cells. Methods: A549 cells were cultured and cells that knockdown of acetyl-CoA carboxylase 1 (ACC1) were obtained by transfection with lentivirus, and cells that knockdown of sterol regulatory element-binding proteins-1 (SREBP-1) were obtained by treated with si-RNA. A549 cells were treated with hypoxia combined with hypoxia inducible factor-1α (HIF-1α) inhibitor PX-478 (25 μmol); Hypoxia combined with linoleic acid (LA) (20 μmol) treated A549 cells with ACC1 knockdown, and A549 cells with SREBP-1 knockdown were treated by hypoxia. Transwell migration assay was used to detect cell migration. Western blot was conducted to detect HIF-1α, ACC1 and epithelial mesenchymal transition (EMT) related proteins, Vimentin, E-Cadherin and SREBP-1; Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was performed to detect the changes of ACC1 and SREBP-1 mRNA in A549 cells after hypoxia and HIF-1α inhibitor PX-478 (25 μmol) treatment. Each experiment was repeated three times. Results: Compared with the normoxic control group, hypoxia promoted the migration of A549 cells (P<0.01), and up-regulated the expressions of ACC1, HIF-1α (all P<0.01) and SREBP-1 (P<0.05). PX-478 (25 μmol) inhibited the migration of A549 cells induced by hypoxia and down-regulated the expression of SREBP-1 (all P<0.05). ACC1 mRNA and SREBP-1 mRNA levels were increased after hypoxia treatment of A549 cells (all P<0.05). The levels of ACC1 mRNA and SREBP-1 mRNA were decreased after A549 cells treated with hypoxia combined with PX-478 (25 μmol) for 24 h (P<0.05, P<0.01). Knockdown of SREBP-1 in A549 cells was obtained by transfection with si-RNA. Transwell migration assay showed the number of cell migration in si-SREBP-1 group was less than that in normoxia control group (P<0.01). The si-SREBP-1 group and the si-NC group were treated with hypoxia. Compared with the control group, the number of cell migration in the si-SREBP-1 group was decreased (P<0.01), however, the difference was not statistically significant compared with the normoxia si-SREBP-1 group (P>0.05). Western blot showed that the expression of ACC1 in the si-SREBP-1 group was lower than that in the control group (P<0.01). Compared with the control group, the expression of ACC1 was decreased after si-SREBP-1 group treated with hypoxia (P<0.01). Knockdown of ACC1 inhibited the migration of A549 cells (P<0.05). After knockdown of ACC1, the migration number of A549 cells under normoxia and 5% O2 conditions had no significant difference (P>0.05). Application of LA under hypoxia condition rescued ACC1-knockdown induced inhibitory effect on hypoxia-promoted A549 cell migration (P<0.05). Conclusion: Hypoxia promotes migration of lung adenocarcinoma A549 cells by regulating fatty acid metabolism through HIF-1α/SREBP-1/ACC1 pathway.


Subject(s)
A549 Cells , Acetyl-CoA Carboxylase , Adenocarcinoma of Lung , Cell Hypoxia/physiology , Cell Line, Tumor , Humans , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit , Lung Neoplasms , RNA/metabolism , RNA, Messenger/metabolism , Sterol Regulatory Element Binding Protein 1/metabolism
2.
Journal of Experimental Hematology ; (6): 1972-1976, 2021.
Article in Chinese | WPRIM | ID: wpr-922234

ABSTRACT

There are more than 150 types of chemical modifications in RNA, mainly methylation, which are widely distributed in all kinds of RNA, including messenger RNA, transfer RNA, ribosomal RNA, non-coding small RNA and long non-coding RNA. In recent years, the identification of RNA methylation modification enzymes and the development of high-throughput sequencing technology at transcriptome level laid a foundation for revealing the expression and function of genes regulated by chemical modification of RNA. In this review, the most recent advances of RNA methylation, especially N6-methyladenosine (m


Subject(s)
Adenosine/metabolism , Hematopoiesis , Humans , Methylation , RNA/metabolism
3.
Protein & Cell ; (12): 792-808, 2020.
Article in English | WPRIM | ID: wpr-880882

ABSTRACT

Over 17 and 160 types of chemical modifications have been identified in DNA and RNA, respectively. The interest in understanding the various biological functions of DNA and RNA modifications has lead to the cutting-edged fields of epigenomics and epitranscriptomics. Developing chemical and biological tools to detect specific modifications in the genome or transcriptome has greatly facilitated their study. Here, we review the recent technological advances in this rapidly evolving field. We focus on high-throughput detection methods and biological findings for these modifications, and discuss questions to be addressed as well. We also summarize third-generation sequencing methods, which enable long-read and single-molecule sequencing of DNA and RNA modification.


Subject(s)
Animals , DNA/metabolism , DNA Methylation , Epigenesis, Genetic , Epigenomics , Humans , RNA/metabolism , Transcriptome
4.
Biol. Res ; 51: 43, 2018. tab, graf
Article in English | LILACS | ID: biblio-983944

ABSTRACT

BACKGROUND: CircRNAs are widespread in plants and play important roles in response to abiotic stresses. Low nitrogen (LN) promotes the growth of plant root system, allowing it to explore more nitrogen. However, whether circRNAs involved in the response to LN stress and the regulation of LN-promoted root growth in wheat remains unclear. METHODS: Two wheat varieties (LH9 and XN979) with contrasting root phenotypes to LN stress were used as materials to identify circRNAs under control and LN conditions by using high-throughput sequencing technology. RESULTS: Six differentially expressed circRNAs (DECs) involved in the common response to LN stress and 23 DECs involved in the regulation of LN-promoted root growth were successfully identified. GO analysis of the DEC-host genes involved in the regulation of LN-promoted root growth showed that GO terms related to biological regulation, responses to stimuli and signalling were significantly enriched. Moreover, seven DECs were predicted to have miRNA binding sites and may serve as miRNA sponges to capture miRNAs from their target genes. CONCLUSIONS: LN stress altered the expression profiles of circRNAs in wheat. This is the first report of LN stress responsive circRNAs in plants. Our results provided new clues for investigating the functions of circRNAs in response to LN stress and in the regulation of LN-promoted wheat root growth.


Subject(s)
Stress, Physiological/physiology , Triticum/growth & development , RNA/isolation & purification , Plant Roots/growth & development , Gene Expression Regulation, Plant/physiology , Nitrogen/metabolism , Triticum/physiology , RNA/metabolism , RNA, Circular
5.
Article in English | WPRIM | ID: wpr-160904

ABSTRACT

DEAD/DExH-box RNA helicases catalyze the folding and remodeling of RNA molecules in prokaryotic and eukaryotic cells, as well as in many viruses. They are characterized by the presence of the helicase domain with conserved motifs that are essential for ATP binding and hydrolysis, RNA interaction, and unwinding activities. Large families of DEAD/DExH-box proteins have been described in different organisms, and their role in all molecular processes involving RNA, from transcriptional regulation to mRNA decay, have been described. This review aims to summarize the current knowledge about DEAD/DExH-box proteins in selected protozoan and nematode parasites of medical importance worldwide, such as Plasmodium falciparum, Leishmania spp., Trypanosoma spp., Giardia lamblia, Entamoeba histolytica, and Brugia malayi. We discuss the functional characterization of several proteins in an attempt to understand better the molecular mechanisms involving RNA in these pathogens. The current data also highlight that DEAD/DExH-box RNA helicases might represent feasible drug targets due to their vital role in parasite growth and development.


Subject(s)
Animals , Eukaryota/enzymology , Gene Expression Regulation , Parasites/enzymology , RNA/metabolism , RNA Helicases/metabolism
6.
Indian J Biochem Biophys ; 2012 Feb; 49(1): 18-24
Article in English | IMSEAR | ID: sea-140214

ABSTRACT

Diabetes is associated with increased formation of advanced glycation end products (AGEs), which have been implicated in micro and macrovascular complications of diabetes. Our earlier reports showed proangiogenic effect of AGE-bovine serum albumin (BSA). In order to understand the mechanism of AGE-mediated angiogenesis, the possibility of involvement of peroxisome prolifeator activated receptor (PPAR) , a ligand activated transcription factor was examined. The angiogenic effect was studied in chick chorio allantoic membrane (CAM) and by analyzing angiogenic markers in human umbilical vein endothelial cells (HUVECs) in culture. The involvement of PPAR was investigated using synthetic PPAR agonist GW 1929 and antagonist GW 9662 and by RT-PCR. In CAM assay, PPAR antagonist GW 9662 reversed the AGE-induced effect on vascularity. In HUVECs in culture, GW 9662 reversed the effect of AGE-BSA and decreased the expression of CD 31, E-Selectin and VEGF. RT-PCR analysis showed that treatment with AGE-BSA caused upregulation of PPAR mRNA levels. The reversal of the effect of AGE on angiogenesis by treatment with PPAR antagonists and up-regulation of PPAR gene in HUVECs treated with AGE-BSA suggested the possible involvement of PPAR -dependent downstream pathway in mediating the angiogenic effect of AGE.


Subject(s)
Angiogenesis Inducing Agents/metabolism , Anilides/pharmacology , Animals , Platelet Endothelial Cell Adhesion Molecule-1/drug effects , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Benzophenones/pharmacology , Cells, Cultured , Chick Embryo , Chorioallantoic Membrane/drug effects , Chorioallantoic Membrane/metabolism , Diabetes Mellitus/metabolism , E-Selectin/metabolism , Glycation End Products, Advanced/pharmacology , Human Umbilical Vein Endothelial Cells/metabolism , PPAR gamma/antagonists & inhibitors , PPAR gamma/antagonists & inhibitors , PPAR gamma/drug effects , PPAR gamma/metabolism , RNA/drug effects , RNA/metabolism , Tyrosine/analogs & derivatives , Tyrosine/pharmacology , Vascular Endothelial Growth Factor A/drug effects , Vascular Endothelial Growth Factor A/metabolism
7.
Article in English | WPRIM | ID: wpr-171961

ABSTRACT

PURPOSE: Retinopathy of prematurity (ROP) is one of the leading causes of blindness, with retinal detachment occurring due to oxygen toxicity in preterm infants. Recently, advances in neonatal care have led to improved survival rates for preterm infants, and ROP has increased in incidence. In the present study, we aimed to determine whether or not resveratrol exhibits protective effects in an animal model of ROP and in primary retinal cell cultures of neonatal rat via nitric oxide (NO)-modulating actions using western blotting and real-time PCR with inducible nitric oxide synthase (iNOS), endothelial NOS (eNOS) and neuronal NOS (nNOS) antibodies and mRNAs. METHODS: In an in vivo oxygen-induced retinopathy (OIR) model, cyclic hyperoxia was induced with 80% O2 for one day and 21% O2 for one day from P1 to P14 in newborn Sprague-Dawley (SD) rats. Resveratrol was injected intravitreally for seven days and rats were sacrificed at P21. In vitro OIR primary retinal cell culture was performed using P0-2 SD rats. Hyperoxia injuries were induced through 100% O2 exposure for six hours. Western blotting and real-time PCR using iNOS, eNOS, nNOS antibodies and primers were performed in the rat model of ROP and the dispersed retinal cell culture. RESULTS: In both in vivo and in vitro OIR, the expression of iNOS antibody and mRNA was increased and of eNOS and nNOS were reduced in the resveratrol-treated group. CONCLUSIONS: In conclusion, resveratrol appeared to exert retinal protective effects via modulation of NO-mediated mechanism in in vivo and in vitro OIR models.


Subject(s)
Analysis of Variance , Animals , Animals, Newborn , Blotting, Western , Disease Models, Animal , Electrophoresis, Polyacrylamide Gel , Humans , Infant, Newborn , Nitric Oxide Synthase/metabolism , Oxygen/toxicity , RNA/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Retina/drug effects , Retinopathy of Prematurity/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Stilbenes/pharmacology
8.
Indian J Biochem Biophys ; 2009 Dec; 46(6): 461-466
Article in English | IMSEAR | ID: sea-135229

ABSTRACT

In the mid-eighties of the last century, extracellular-proteolipid complexes have been identified in tumor patients and circulating RNA was suggested to represent a specific secretory product of cancer cells. The presence of specific types of RNA in a variety of cancer types proved to be useful in cancer diagnosis. It has been suggested that extracellular RNA and DNA are not inert molecules, but contain biological activities. Recent data have demonstrated that extracellular RNA is likely to present the up to now undefined “natural foreign surface”, serving as an initiating factor in blood coagulation in vivo. Yet, extracellular RNA seems to have even more functions. Investigations on blood-brain-barrier have shown that extracellular RNA mediates endothelial permeability. Ample success has been achieved in administrating RNase in different animal models of vascular diseases, thereby significantly delaying thrombus formation and reducing cerebral edema formation with neuroprotection in acute stroke models. Furthermore, extracellular mammalian RNA was found to decrease tumor yield in a murine model system, suggesting that extracellular RNA might trigger immune response. Finally, extracellular nucleic acids were identified as danger signals involved in innate immunity related to neutrophil-mediated bacterial killing and haemocyte activation and coagulation in the insects. Thus, a new area of research on extracellular RNA functions with promising future perspectives just started in the field of inflammation and immunity.


Subject(s)
Animals , Blood Coagulation , Extracellular Space/enzymology , Extracellular Space/metabolism , Humans , Immunity, Innate , Inflammation/blood , Inflammation/enzymology , Inflammation/immunology , Inflammation/pathology , RNA/metabolism , Ribonucleases/metabolism
9.
Article in English | IMSEAR | ID: sea-34847

ABSTRACT

Human telomerase consisting of telomerase RNA template (hTR) and telomerase reverse transcriptase (hTERT) provides a mechanism for synthesis of telomere repeats that prolongs life span of cells. Telomerase activity is present in germ-line and malignant tumor cells but not in most normal human somatic cells. This study determined hTERT mRNA level in tissue samples from patients with gastrointestinal tract (GI) cancers. Tissue samples were obtained from 22 GI cancer patients, 3 gastrointestinal stomal tumors (GIST) and 25 corresponding non-cancerous tissues. hTERT expression was determined by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) using Taqman probe, hTERT mRNA was detected in 12 of 22 cancerous tissue samples. Six of 8 tissue samples obtained from patients with hepatocellular carcinoma and cholangiocarcinoma were positive for hTERT. However, hTERT mRNA was not detected in GIST and non-cancerous tissues. These results suggest that hTERT may be an effective target for cancer therapies to treat many type of GI cancers including cholangiocarcinoma and hepatocellular carcinoma.


Subject(s)
Carcinoma, Hepatocellular/enzymology , Cholangiocarcinoma/enzymology , Gastrointestinal Neoplasms/enzymology , Humans , RNA/metabolism , Telomerase/metabolism
10.
Article in English | WPRIM | ID: wpr-634590

ABSTRACT

In order to investigate the role of Twist gene in the metastasis of hepatocellular carcinoma (HCC), total RNA was respectively extracted from three HCC cell strains with different metastatic potentials, HepG2, MHCC-97L and MHCC-97H. The first strand cDNA was synthesized by reverse transcription, which was then used as template to perform fluorescent quantitative polymerase chain reaction (FQ-PCR). The quantity of Twist gene expression was normalized by that of the housekeeping gene, GAPDH for each sample. ANOVA was used to estimate the relationship between Twist gene and metastasis potential of HCC. The results showed that the normalized initial cDNA concentrations of Twist gene in HepG2, MHCC-97L and MHCC-97H were (9.45+/-0.25)x10(-4), (1.82+/-0.41)x10(-3), (3.06+/-0.62)x10(-3), respectively. FQ-PCR revealed significant differences in the expression level of Twist among HCC cell strains with different metastatic potentials. It was concluded that high expression level of Twist was closely associated with more aggressive behaviors of HCC. Twist provides a novel indicator for HCC metastasis.


Subject(s)
Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor , DNA, Complementary/metabolism , Dose-Response Relationship, Drug , Gene Expression Regulation, Neoplastic , Liver Neoplasms/metabolism , Neoplasm Metastasis , Nuclear Proteins/biosynthesis , Polymerase Chain Reaction , RNA/metabolism , Twist-Related Protein 1/biosynthesis
11.
Indian J Biochem Biophys ; 2007 Jun; 44(3): 152-6
Article in English | IMSEAR | ID: sea-28486

ABSTRACT

The effect of inositol supplementation on glucose derepression, invertase secretion and SUC2 gene expression in Saccharomyces sp. W4 was studied. Invertase secretion was repressed, when the yeast cells, grown the synthetic medium without inositol (I(-) medium) contained more than 0.2% (w/v) initial concentration of glucose. However, in the same medium plus inositol (I(+) medium, inositol conc. 100 microg/100 ml), invertase secretion was repressed only at glucose concentrations higher than 2.0% (w/v). Results showed that secreted invertase activity increased only in the I+ medium, whereas intracellular invertase activity remained constant in both media during the cell, growth. The mRNA encoding secreted invertase was higher in the glucose-derepressed cells grown in the I(+) medium than in the glucose-repressed cells grown in the I(-) medium. Similarly, phosphatidylinositol (PI) content was significantly higher in the cells grown in the I(+) medium than in the I(-) medium. These results indicated that PI might be involved in the glucose derepression, invertase secretion and SUC2 gene expression at the transcriptional level in the yeast.


Subject(s)
Cell Culture Techniques , Culture Media , Dose-Response Relationship, Drug , Gene Expression Regulation, Fungal , Glucose/metabolism , Inositol/metabolism , Phospholipids/metabolism , RNA/metabolism , RNA, Fungal/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Saccharomyces/metabolism , Saccharomyces cerevisiae Proteins/biosynthesis , Time Factors , beta-Fructofuranosidase/biosynthesis
12.
Article in English | WPRIM | ID: wpr-110322

ABSTRACT

Barrett's esophagus is a premalignant condition of esophageal adenocarcinoma. Inducible nitric oxide synthase (iNOS) is induced by cytokines and can generate locally high concentrations of nitric oxide (NO), whose metabolites can mediate genotoxicity and influence multistage carcinogenesis by causing DNA damage. Therefore, we evaluated the immunolocalization and expression of iNOS in surgically induced rat Barrett's esophagus. Esophagoduodenal anastomosis was performed in rats for inducing reflux of duodenal contents. Rats were killed at postoperative 10, 20, 30 and 40 weeks. We examined histologic changes and iNOS expression in esophagus by immunohistochemistry and reverse transcription-poly-merase chain reaction. Eighty six percent of experimental rats showed Barrett's esophagus above esophagoduodenal junction. iNOS immunoreactivity was clearly observed in the epithelial cells of Barrett's esophagus, predominantly at the apical surface of epithelial cells. Cytoplasmic staining was also seen only in atypical Barrett's esophagus. iNOS mRNA was detected only in the lower esophagus of experimental group. In conclusion, this study suggests that iNOS has some roles on Barrett's esophagus formation.


Subject(s)
Anastomosis, Surgical , Animals , Barrett Esophagus/enzymology , Cytoplasm/metabolism , DNA Damage , Disease Models, Animal , Duodenum/enzymology , Esophagus/metabolism , Immunohistochemistry , Male , Models, Anatomic , Neoplasms, Experimental/pathology , Nitric Oxide/metabolism , Nitric Oxide Synthase/biosynthesis , RNA/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
14.
Article in English | IMSEAR | ID: sea-21445

ABSTRACT

BACKGROUND & OBJECTIVES: Echovirus 11 (ECV11) is one of the most frequent non-polio enteroviruses isolated from stool samples of children with acute flaccid paralysis in north India. The present work was undertaken to study the sequence variability in the 440 bp of 5'-non-translated region of ECV11 genome using heteroduplex mobility assay (HMA). METHODS: Twelve ECV11 isolates were studied for sequence variability in the 5'-non-translated region (5'NTR) using the HMA followed by nucleotide sequencing. HMA was used to determine sequence diversity between Indian ECV11 isolates and prototype Gregory strain. HMA results were confirmed by 5'NTR nucleotide sequencing of five Indian ECV11 isolates. RESULTS: HMA results showed high genomic diversity between the prototype Gregory strain and Indian ECV11 isolates. All isolates were grouped into five different types of heteroduplex mobility patterns with respect to Gregory strain. A 440 bp 5'NTR fragment of five ECV11 isolates representing different heteroduplex patterns, was sequenced. The sequence alignment showed that 5'NTR of Indian isolates was different from prototype Gregory strain and identical to the ECV11 isolates of Finland and Hungary. Phylogenetic analysis including ECV11 isolate sequences from different parts of the world showed that Indian ECV11 isolates represented a different subgroup. INTERPRETATION & CONCLUSION: The results of the present study suggested that the HMA could be successfully used as a preliminary screening method for sequence variability determination of enterovirus field isolates. The sequence data generated on ECV11 isolates from India will be useful for future studies of endemic genotypes of echovirus.


Subject(s)
5' Untranslated Regions , Child, Preschool , DNA/genetics , DNA, Complementary/metabolism , Enterovirus B, Human/genetics , Female , Genetic Techniques , Genome, Viral , Humans , India , Infant , Male , Nucleic Acid Heteroduplexes/genetics , Phylogeny , Poliovirus Vaccine, Oral/pharmacology , Polymerase Chain Reaction , RNA/metabolism , RNA, Viral , Reverse Transcriptase Polymerase Chain Reaction
15.
Article in English | IMSEAR | ID: sea-20363

ABSTRACT

BACKGROUND & OBJECTIVES: Hepatitis C virus (HCV), an important cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma, shows a considerable genetic heterogeneity among hepatitis C virus isolates from all over the world. At least six main groups of sequence variants are recognized. The natural history of disease and response to treatment may be related to the genotype of HCV in a particular patient. Antigenic differences between genotypes also have implications for optimal design of serological sequencing and confirmatory assays for HCV. The present study was undertaken with the objective to find out various genotypes of hepatitis C virus prevalent in Indian patients with chronic hepatitis C infection. METHODS: Thirty six consecutive newly diagnosed patients with chronic hepatitis C infection were included in the study. HCV RNA was extracted from the serum by standard guanidinium thiocyanate method. Following reverse transcription and amplification, the HCV genotypes were determined by line probe assay (INNO-LiPA HCV II). RESULTS: Of the 36 patients, genotype 3 was found in 24 (66.6%). Of these 24 patients, 3a was seen in 5 patients (13.8%), 3b in two (5.5%) and mixed subtype 3a and 3b in 17 patients (47.2%). Genotype 1 was found in 5 patients (13.8%), with 1b in 1 and 1a in rest four cases. Two patients (5.5%) were infected with genotype 2 (subtype 2a and mixed subtype 2a, 2b respectively). One (2.7%) was infected with genotype 4 (4a). Mixed genotype infection was found in 4 patients (11.1%). INTERPRETATION & CONCLUSION: The present findings showed that genotype 3 of hepatitis C virus was the most prevalent genotype in patients with chronic hepatitis C in this part of India.


Subject(s)
DNA, Viral/genetics , Disease Progression , Genotype , Guanidines/pharmacology , Hepacivirus/genetics , Hepatitis Antibodies , Hepatitis C/genetics , Hepatitis C, Chronic/genetics , Humans , India , Polymerase Chain Reaction , RNA/metabolism , RNA, Viral , Thiocyanates/pharmacology
16.
Article in English | WPRIM | ID: wpr-124477

ABSTRACT

Transforming growth factor (TGF)-beta1 is an important fibrogenic factor that is involved in the pathogenesis of diabetic nephropathy. We evaluated the effect of circular antisense TGF-beta1 oligodeoxynucleotides (ODNs) on the TGF-beta1 expression in the rat mesangial cell culture and in streptozotocin (STZ)-induced diabetic rats. Circular antisense TGF-beta1 ODNs were found to be stable in rat serum, significantly decreasing TGF-beta1 mRNA expression compared with linear antisense ODNs in the rat mesangial cell culture. Circular antisense TGF-beta1 ODNs were introduced into the tail vein of normal rats using hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer method and were confirmed to be delivered effectively into the kidney, liver, lungs, and spleen. To inhibit the overexpression of TGF-beta1 in diabetic kidneys, we introduced circular antisense TGF-beta1 ODNs into the STZ-induced diabetic rats. On day 13 after circular antisense TGF-beta1 ODNs injection, TGF-beta1 mRNA and protein expression markedly decreased and urinary TGF-beta1 excretion rate also dropped in the circular antisense TGF-beta1 ODNs-treated diabetic rats. These results suggest that circular antisense TGF-beta1 ODNs may be a useful tool for developing new therapeutic application for progressive diabetic nephropathy.


Subject(s)
Animals , Blood Glucose/metabolism , Cells, Cultured , Diabetes Mellitus, Experimental/therapy , Enzyme-Linked Immunosorbent Assay , Gene Transfer Techniques , Immunohistochemistry , Liposomes/chemistry , Male , Microscopy, Confocal , Oligonucleotides, Antisense/metabolism , RNA/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Streptozocin , Time Factors , Transfection , Transforming Growth Factor beta/genetics
17.
Article in English | WPRIM | ID: wpr-634202

ABSTRACT

Telomerase activity and the expression of telomerase subunits (for example, telomerase reverse transcriptase and telomerase associated protein 1 and telomerase RNA component) of peripheral white blood cells were detected in the patients with acute myelogenous leukaemia (AML) and the correlation between telomerase activity and the expression of telomerase subunits was observed. In 94 peripheral white blood cells from 18 healthy volunteers and 76 patients with AML, including 31 AML at initial presentation, 24 at relapse and 21 at complete remission, the telomerase activity and telomerase subunits mRNA or RNA were detected by PCR-ELISA and RT-PCR respectively. The results showed that the positive rate of telomerase from patients with AML at initial presentation, at relapse and at complete remission was 74.1%, 79.2% and 4.8% respectively. The positive rate of telomerase reverse transcriptase mRNA from healthy volunteers, AML at initial presentation, AML at relapse and AML at complete remission was 5.6%, 80.6%, 83.3% and 9.5% respectively. The positive rate of telomerase associated protein 1 mRNA and telomerase RNA component in all samples were 100%. It was suggested that the up-regulation of telomerase activity and the expression of telomerase reverse transcriptase is correlated closely with the occurrence and relapse of AML, so telomerase activity and the expression of telomerase reverse transcriptase may be used to estimate the curative effect and predict relapse of AML. Moreover, the up-regulation of telomerase activity is correlated with the expression of telomerase reverse transcriptase significantly.


Subject(s)
Carrier Proteins/metabolism , DNA-Binding Proteins , Leukemia, Myeloid, Acute/enzymology , RNA/metabolism , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Telomerase/metabolism
18.
Article in English | WPRIM | ID: wpr-221862

ABSTRACT

The present study evaluated the importance of ovarian functions and the renin-angiotensin system in the progression of the right ventricular (RV) hypertrophy. Female Sprague-Dawley rats were bilaterally ovariectomized (Ovx) and injected with monocrotaline (MCT, 60 mg/kg, sc). Four weeks after MCT-treatment, only the male and Ovx female rats showed marked RV hypertrophy. The hypertrophied RV of the male-MCT and Ovx-MCT rats exhibited remarkably elevated renin mRNA levels. Gene expression levels of angiotensinogen, TGF-beta1, and endothelin-1 in the hypertrophied RV also increased, but to the less degree than did the renin mRNA. To investigate beneficial effects of estrogen or enalapril on progression of the pulmonary hypertension and RV hypertrophy, histological changes of the lung and heart were examined. Sham-MCT female rats showed histological changes indicating pulmonary hypertension without RV hypertrophy. In contrast, Ovx-MCT rats showed marked RV hypertrophy with pathological changes, denoting severe pulmonary and myocardial injuries. Estrogen-or enalapril-treated Ovx-MCT rats did not show RV hypertrophy, and showed remarkably ameliorated ultrastructural changes in the lung and RV. These results from this rat model suggest that both estrogen and inhibition of the renin-angiotensin system have protective functions against the development of the pulmonary hypertension and cardiac remodeling.


Subject(s)
Angiotensin-Converting Enzyme Inhibitors/pharmacology , Angiotensinogen/biosynthesis , Animals , Body Weight/drug effects , Densitometry , Disease Progression , Enalapril/pharmacology , Endothelin-1/biosynthesis , Estrogens/pharmacology , Female , Hypertrophy, Right Ventricular/chemically induced , Male , Microscopy, Electron , Monocrotaline/pharmacology , Ovariectomy , RNA/metabolism , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Renin/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Sex Factors , Transforming Growth Factor beta/biosynthesis , Ventricular Remodeling
19.
Article in English | WPRIM | ID: wpr-10318

ABSTRACT

Telomeres serve a critical role in maintenance of genomic stability in all eukaryotes, from yeast to human. The maintenance of telomeres is achieved by the telomerase complex, which is largely composed of telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC). A variety of mouse models have provided valuable insights into the relationship between the telomerase complex and telomere dysfunction at the organismal level and helped understand their biological significance in human. Recently, in addition to its role in maintenance of the telomeres, novel functions of the telomerase complex have been emerging. In this review, studies of all gene-targeted or transgenic mouse models so far generated for telomerase and telomere biology are comprehensively described, and potential novel functions of telomerase are briefly discussed


Subject(s)
Animals , Cellular Senescence/physiology , Mice , Mice, Knockout , Mice, Transgenic , Models, Animal , RNA/metabolism , Telomerase/metabolism , Telomere/metabolism
20.
Braz. j. med. biol. res ; 34(3): 295-300, Mar. 2001. ilus
Article in English | LILACS | ID: lil-281609

ABSTRACT

Recent research has shown that receptor-ligand interactions between surfaces of communicating cells are necessary prerequisites for cell proliferation, cell differentiation and immune defense. Cell-adhesion events have also been proposed for pathological conditions such as cancer growth, metastasis, and host-cell invasion by parasites such as Trypanosoma cruzi. RNA and DNA aptamers (aptus = Latin, fit) that have been selected from combinatorial nucleic acid libraries are capable of binding to cell-adhesion receptors leading to a halt in cellular processes induced by outside signals as a consequence of blockage of receptor-ligand interactions. We outline here a novel approach using RNA aptamers that bind to T. cruzi receptors and interrupt host-cell invasion in analogy to existing procedures of blocking selectin adhesion and function in vitro and in vivo


Subject(s)
Humans , Cell Adhesion Molecules/physiology , DNA/metabolism , RNA-Binding Proteins/metabolism , RNA/metabolism , Trypanosoma cruzi , Cell Adhesion , Chagas Disease/parasitology , DNA/chemistry , DNA/isolation & purification , Host-Parasite Interactions , Integrins/metabolism , L-Selectin/analysis , P-Selectin/analysis , Protozoan Proteins/chemistry , Protozoan Proteins/metabolism , RNA/chemistry , RNA/isolation & purification , Trypanosoma cruzi/metabolism
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