1.A Novel Type of Non-coding RNA, nc886, Implicated in Tumor Sensing and Suppression.
Genomics & Informatics 2015;13(2):26-30
nc886 (=vtRNA2-1, pre-miR-886, or CBL3) is a newly identified non-coding RNA (ncRNA) that represses the activity of protein kinase R (PKR). nc886 is transcribed by RNA polymerase III (Pol III) and is intriguingly the first case of a Pol III gene whose expression is silenced by CpG DNA hypermethylation in several types of cancer. PKR is a sensor protein that recognizes evading viruses and induces apoptosis to eliminate infected cells. Like viral infection, nc886 silencing activates PKR and induces apoptosis. Thus, the significance of the nc886:PKR pathway in cancer is to sense and eliminate pre-malignant cells, which is analogous to PKR's role in cellular innate immunity. Beyond this tumor sensing role, nc886 plays a putative tumor suppressor role as supported by experimental evidence. Collectively, nc886 provides a novel example how epigenetic silencing of a ncRNA contributes to tumorigenesis by controlling the activity of its protein ligand.
Apoptosis
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Carcinogenesis
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DNA
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Epigenomics
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Immunity, Innate
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Protein Kinases
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RNA Polymerase III
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RNA, Untranslated*
2.A Case of Renal Crisis in a Korean Scleroderma Patient with Anti-RNA polymerase I and III Antibodies.
Eun Ha KANG ; Churl Hyun IM ; Su Ho KIM ; Jae Rak CHUNG ; Eun Young LEE ; Dong Jo KIM ; Eun Bong LEE ; Yeong Wook SONG
Journal of Korean Medical Science 2006;21(6):1121-1123
Scleroderma (SSc) renal crisis has been reported to be associated with anti-RNA polymerase I and III (RNAP I/III) antibodies in Caucasians and the Japanese. However, no report is available for Korean SSc patients. Here, we describe the case of a 65-yr-old female SSc patient who developed renal crisis and whose serum contained anti-RNAP I/III antibodies. She was finally diagnosed as having diffuse cutaneous SSc based on skin thickening proximal to the elbows and knees. Sudden hypertension, oliguria, and pulmonary edema were features of her renal crisis. Despite the use of captopril and adequate blood pressure control, her renal function deteriorated. Subsequent renal biopsy findings showed severe fibrinoid necrosis with luminal obliteration in interlobar arteries and arterioles consistent with SSc renal crisis. Serum anti-RNAP I/III antibodies were detected by radioimmunoprecipitation. This is the first report of a renal crisis in a Korean SSc patient with RNAP I/III antibodies.
Scleroderma, Systemic/*complications/enzymology/*immunology
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RNA Polymerase III/*immunology
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RNA Polymerase I/*immunology
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Korea
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Kidney Failure, Acute/*complications/enzymology/*immunology
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Humans
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Female
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Critical Care
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Autoantibodies/*blood
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Aged
3.Inhibition of COL1A1 and COL3A1 expression by RNA interference in human skin fibroblasts.
Qiong WANG ; Zhen-Hui PENG ; Wan-Juan WANG
Journal of Southern Medical University 2008;28(1):1-6
OBJECTIVETo suppress COL1A1 and COL3A1 gene expressions in human skin fibroblasts (HSFs) by means of RNA interference (RNAi).
METHODSThree small interfering RNA (siRNA) expression cassette (SEC) sequences were designed for each of the COL1A1 and COL3A1 gene sequences available in GenBank. The synthesized SECs capable of effective gene suppression were transfected into cultured HSFs, either after cloning into the expression vector or mediated by Lipofectamine 2000, and the suppression of the target genes at both mRNA and protein levels was determined by quantitative fluorescence RT-PCR and Western blotting, respectively.
RESULTSTransfection of the SECs into HSFs resulted in specific depression of COL1A1 and COL3A1 expressions (down to 5.00% and 6.48%, respectively). The expression vector-mediated RNAi established a HSF cell line with persistent gene knockdown for over 30 days (to 25.21% and 22.12%, respectively).
CONCLUSIONCOL1A1 and COL3A1 gene expressions can be specifically and efficiently inhibited in HSFs by either liposome- or vector-mediated SEC transfection.
Blotting, Western ; Cells, Cultured ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Fibroblasts ; cytology ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Skin ; cytology ; Transfection ; methods
4.Comparative Study of the Effects of Trabecular Meshwork Outflow Drugs on the Permeability and Nitric Oxide Production in Trabecular Meshwork Cells.
Korean Journal of Ophthalmology 2017;31(5):452-459
PURPOSE: To compare the effects of the barrier function in human trabecular meshwork (TM) cells monolayer and the production of nitric oxide (NO) between trabecular outflow drugs, Rho-associated kinase (ROCK) inhibitors, adenosine, and statin. METHODS: Primary cultured TM cells were exposed to 10 or 25 µM Y-27632, 0.1 or 1 µM N6-cyclohexyladenosine (CHA), or 15 or 30 µM simvastatin for 24 hours. NO production and expression of endothelial nitric oxide synthase mRNA were measured by Griess assay and reverse transcription polymerase chain reaction, respectively. Barrier functions of the TM cell monolayer were measured by carboxyfluorescein and trans-endothelial electrical resistance. The expression of matrix metalloproteinase-2 mRNA was assessed with reverse transcription polymerase chain reaction. RESULTS: In TM cells, treatment with each drug increased endothelial nitric oxide synthase mRNA expression. Treatment with 25 µM Y-27632 and 1.0 µM CHA increased NO production significantly (p = 0.035 and p = 0.043, respectively). Treatment with each drug increased the permeability (all p = 0.001) and decreased the trans-endothelial electron resistance of the TM cell monolayer. Treatment with 0.1 µM and 1.0 µM CHA significantly increased matrix metalloproteinase-2 mRNA expression, but simvastatin inhibited its expression. CONCLUSIONS: Since treatment with ROCK inhibitor more greatly increased NO production and permeability than did adenosine or statin, ROCK inhibitor seems to be more effective for lowering intraocular pressure.
Adenosine
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Electric Impedance
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Humans
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Hydroxymethylglutaryl-CoA Reductase Inhibitors
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Intraocular Pressure
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Matrix Metalloproteinase 2
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Nitric Oxide Synthase Type III
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Nitric Oxide*
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Permeability*
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Polymerase Chain Reaction
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Reverse Transcription
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rho-Associated Kinases
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RNA, Messenger
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Simvastatin
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Trabecular Meshwork*
5.Evaluation of the Virus-elimination Efficacy of Nanofiltration (Viresolve NFP) for the Parvovirus B19 and Hepatitis A Virus.
Deok Ja OH ; Yoo La LEE ; Jae Won KANG ; So Yong KWON ; Nam Sun CHO ; In Seop KIM
The Korean Journal of Laboratory Medicine 2010;30(1):45-50
BACKGROUND: The safety of plasma derivatives has been reinforced since 1980s by variable pathogen inactivation or elimination techniques. Nucleic acid amplification test (NAT) for the source plasma has also been implemented worldwide. Recently nanofiltration has been used in some country for ensuring safety of plasma derivatives to eliminate non-enveloped viruses such as parvovirus B19 (B19V) and hepatitis A virus (HAV). We evaluated the efficacy of nanofiltration for the elimination of B19V and HAV. METHODS: To verify the efficacy of nanofiltration, we adopted a 20 nm Viresolve NFP (Millipore, USA) in the scaling down (1:1,370) model of the antithrombin III production. As virus stock solutions, we used B19V reactive plasma and porcine parvovirus (PPV) and HAV obtained from cell culture. And 50% tissue culture infectious dose was consumed as infectious dose. The methods used to evaluate the virus-elimination efficacy were reverse-transcriptase polymerase chain reaction for B19V and the cytopathic effect calculation after filtration for PPV and HAV. RESULTS: B19V was not detected by RT-PCR in the filtered antithrombin III solutions with initial viral load of 6.42x10(5) IU/mL and 1.42x10(5) IU/mL before filtration. The virus-elimination efficacy of nanofiltration for PPV and HAV were > or =10(3.32) and > or =10(3.31), respectively. CONCLUSIONS: Nanofiltration would be an effective method for the elimination of B19V and HAV. It may be used as a substitute for NAT screening of these viruses in source plasma to ensure safety of plasma derivatives in Korea.
Antithrombin III/isolation & purification
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DNA, Viral/analysis
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Filtration/*methods
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Hepatitis A virus/genetics/*isolation & purification
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Humans
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Nanotechnology/*methods
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Parvovirus B19, Human/genetics/*isolation & purification
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RNA, Viral/analysis
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Reverse Transcriptase Polymerase Chain Reaction
6.Effects of Metformin on Penile Expression of Nitric Oxide Synthase in OLETF Rats.
Phil Hyun SONG ; In Soo BAE ; Yong Woon KIM ; Ki Hak MOON
Korean Journal of Urology 2009;50(2):179-185
PURPOSE: It is well known that metformin, an oral biguanide insulinsensitizing agent, targets AMP-activated protein kinase (AMPK), which activates endothelial nitric oxide synthase (eNOS), which may be associated with erectile dysfunction. The aim of this study was to evaluate whether metformin activates eNOS in the penile tissue of a genetically obese rat model. MATERIALS AND METHODS: We measured phospho-eNOS levels and eNOS expression in the penile tissue of Otsuka Long Evans Tokushima Fatty (OLETF) rats after 3 days of treatment with metformin (150 or 300 mg) to evaluate whether metformin activates eNOS in the penile tissue of this genetically obese rat model. Seven-month-old OLETF rats were used, and eNOS expression was analyzed by real-time polymerase chain reaction (PCR). The experimental groups were compared with use of the Kruskal-Wallis or Mann-Whitney test. RESULTS: Seven-month-old OLETF rats had severe visceral fat deposition and elevated serum leptin concentrations. eNOS expression analyzed by real-time PCR was lower in the penile tissue of OLETF rats than in Long Evans Tokushima Otsuka (LETO) rats. Short-term treatment with metformin did not change visceral fat mass or serum leptin levels. However, metformin treatment increased eNOS mRNA expression in the penile tissue as determined by real-time PCR. The levels of phospho-eNOS and phospho-AMPK (pAMPK) in penile tissue revealed a dose-dependent tendency to increase with metformin treatment; however, there was no statistical difference by Kruskal-Wallis test among the experimental groups. The pAMPK level was dose-dependently elevated in the soleus by metformin treatment. There was no significant change in pSTAT3 by metformin treatment in the soleus. CONCLUSIONS: Metformin activates eNOS mRNA expression in the penile tissue of OLETF rats. Further study on the relation between erectile function and eNOS levels is needed for clinical application.
AMP-Activated Protein Kinases
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Animals
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Erectile Dysfunction
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Intra-Abdominal Fat
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Leptin
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Male
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Metformin
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Nitric Oxide
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type III
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Penis
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Rats
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Rats, Inbred OLETF
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Real-Time Polymerase Chain Reaction
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RNA, Messenger
7.Protein and mRNA expression of nitric oxide synthase isoforms I and III in rat penile spongium tissues.
Jun-Ping SHI ; Yong-Mei ZHAO ; Gang QI
National Journal of Andrology 2003;9(9):663-668
OBJECTIVETo investigate protein and mRNA expression of nitric oxide synthase (NOS) isoforms I and III in rat penile tissues.
METHODSPenile tissues from male SD rats were investigated. The protein expression of NOS I and III in rat penis was confirmed by immunoprecipitation and Western blot. Gene expression of both NOS I and III was identified by reverse transcription-polymerase chain reaction.
RESULTSThe mRNA and protein products of NOS I and III are both expressed in normal rat penile spongium tissues. But, NOS III mRNA and protein products are more highly expressed than those of NOS.
CONCLUSIONSThese findings support the possibility that penile erection is regulated by different NOS isoforms in rat penile tissues.
Animals ; Blotting, Western ; Male ; Nitric Oxide Synthase ; analysis ; genetics ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type III ; Penis ; enzymology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction
8.Expression of COL1a1 and COL3a1 in the capsule of children with developmental dislocation of the hip.
En-Bo WANG ; Qun ZHAO ; Lian-Yong LI ; Li-Wei SHI ; Hong GAO
Chinese Journal of Contemporary Pediatrics 2008;10(4):493-496
OBJECTIVEThe etiology of developmental dislocation of the hip (DDH) remains uncertain, but some research has shown that this disorder is closely related to hip joint laxity. This study examined the expression of collagens type I and III mRNA and protein in the hip capsule of children with DDH in order to investigate the roles of collagens type I and III in hip joint laxity.
METHODSNine children with DDH and nine age and gender-matched normal children (control group) were enrolled. Semiquantitative RT-PCR method was used to detect mRNA expression of COL1a1 and COL3a1 in the hip capsule. Western-Blot method was used to detect protein expression of COL1a1 and COL3a1 in the hip capsule. The quantitative analysis of the COL1a1 and COL3a1 was performed by professional image software and the results were analyzed with standard statistical methods.
RESULTSmRNA and protein expression of COL1a1 in the DDH group was significantly lower than that in the control group (P<0.01). Compared with the control group, COL1a3 mRNA expression in the DDH group decreased significantly (P<0.01), but COL1a3 protein expression was not significantly different.
CONCLUSIONSThe decreased collagen I mRNA and protein expression in the hip capsule might contribute to hip joint laxity in children with DDH. Collagen type III may not be associated with hip joint laxity in DDH.
Blotting, Western ; Child ; Child Development ; Child, Preschool ; Collagen Type I ; analysis ; genetics ; Collagen Type III ; analysis ; genetics ; Female ; Hip Dislocation ; metabolism ; Humans ; Infant ; Male ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction
9.Decreased expression of DICER1 in gastric cancer.
Zhi-hong ZHENG ; Xiu-ju SUN ; Wei-neng FU ; Yi GUAN ; Feng GAO ; Ying WANG ; Kai-lai SUN
Chinese Medical Journal 2007;120(23):2099-2104
BACKGROUNDThe role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.
METHODSTo detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.
RESULTSIn 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).
CONCLUSIONSDICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.
Blotting, Western ; DEAD-box RNA Helicases ; analysis ; genetics ; physiology ; Endoribonucleases ; analysis ; genetics ; physiology ; Epigenesis, Genetic ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Ribonuclease III ; Stomach Neoplasms ; chemistry ; etiology ; genetics
10.Association of microRNA-related genes (DROSHA, DICER1 and GEMIN4) polymorphisms with T-cell lymphoma prognosis.
Xiaobo TIAN ; Bo ZHANG ; Xi LI ; Kan ZHAI ; Jian XU ; Jiang CHANG ; Yan QIAO ; Yuling ZHOU ; Liming HUANG ; Jieping CHEN
Chinese Journal of Hematology 2014;35(5):408-413
OBJECTIVETo analyze the association of micoRNA-related genes DROSHA single nucleotide polymorphisms (SNP) rs10719 and rs6877842, DICER1 rs3742330and GEMIN4 rs3744741 with prognosis of T-cell lymphoma.
METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to determine the genotypes of the above 4SNPs and their associations with complete remission (CR) rate and overall survival (OS) in 163 patients with TCL.
RESULTSPatients carrying the rs6877842 CG genotype had a significantly higher CR rate compared with those carrying the CC genotype (OR=0.07, 95% CI 0.01-0.72, P=0.026); the same for patients carrying the DICER1 rs3742330 GG genotype compared with those carrying the GA genotype (OR=0.15, 95% CI 0.02-0.97, P=0.047) or the AA genotype (OR=0.11, 95% CI 0.02-0.71, P=0.020). In addition, patients with the DICER1 rs3742330 GG genotype had a significantly improved OS compared with those carrying the GA (HR=9.02, 95% CI 1.22-66.92, P=0.031) or AA genotype (HR=8.77, 95% CI 1.19-64.67, P=0.033). The other two SNPs of rs10719 and rs3744741 had no significant association with CR or OS.
CONCLUSIONDROSHA rs6877842 and DICER1 rs3742330 were independent factors for TCL CR, and DICER1 rs3742330 was also an independent prognostic factor for TCL OS.
DEAD-box RNA Helicases ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lymphoma, T-Cell ; diagnosis ; genetics ; MicroRNAs ; genetics ; Minor Histocompatibility Antigens ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Prognosis ; Ribonuclease III ; genetics ; Ribonucleoproteins, Small Nuclear ; genetics