• Title/Summary/Keyword: Gene Medicine

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Association analysis of a polymorphism of the angiotensin I-converting enzyme gene and angiotensin II Type 1 receptor gene in Korean population

  • Yang, Young-Mok;Park, Jong-Hwan;Lee, Hyun-Young;Moon, Eon-Soo
    • Journal of Genetic Medicine
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    • v.2 no.1
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    • pp.27-30
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    • 1998
  • Previously, we made a study report on the genotype distribution and the gene frequency of angiotesin I-converting enzyme (ACE) in Korean population, and on the association between hypertension and genetic variance of ACE. This time, we have investigated a rapid mismatch-PCR/RFLP assays for the variant of the angiotesin II type 1 receptor ($AT_1R$) gene (an $A{\rightarrow}C$ transversion at position 1166 of $AT_1R$ gene), a mutation which may interact with the ACE polymorphism in the determining of risk of myocardial infarction. The genotype distributions of Koreans' angiotensin II type 1 receptor gene were AA (66.3%):AC (28.1%):CC (5.6%), thus the AA genotype was most numerous, and the allele frequency was A:C = 0.803:0.197. Genotype distributions were shown as AA (76.8%):AC (20.9%):CC (2.3%), the allele frequency was A:C = 0.872:0.128 in the male group, and AA (47.4%):AC (41.0%):CC (11.6%), A:C = 0.679:0.321 in the female group. Differences were highly significant between the male and female groups (p<0.0001). Genotype distributions between angiotensin II type 1 receptor gene and angiotensin converting enzyme gene showed that there is no significance between $AT_1R$ genotypes and ACE genotypes in total subjects (p>0.05).

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The Effect of Galkunhwanglyeonhwanggum-tang (GGT) and Sopunghwalhyeol-tang (SPT) on Gene Expression Levels of MCP-1, ICAM-1, VCAM-1, and eNOS in HUVECs (갈근황연황금탕(葛根黃蓮黃芩湯)과 소풍활혈탕(疎風活血湯)이 HUVEC 내에 MCP-1, ICAM-1, VCAM-1 and eNOS의 유전자 발현량에 대해 미치는 영향)

  • Jeong, Hyun-jin;Jeon, Sang-yoon;Jang, Hye-yeon;Kim, Min-wook
    • The Journal of Internal Korean Medicine
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    • v.41 no.4
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    • pp.668-675
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    • 2020
  • Objectives: The aim of this study was to compare the effects of Galkunhwanglyeonhwanggum-tang (GGT), and Sopunghwalhyeol-tang (SPT) on gene expression of MCP-1, ICAM-1, VCAM-1, and eNOS in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were treated with GGT and SPT at concentrations of 50, 100, and 200 ㎍/mL. Gene expression of MCP-1, ICAM-1, VCAM-1, and eNOS in HUVECs was analyzed by the polymerase chain reaction (PCR), and electrophoresis was performed to verify the gene expression level. Results: 1. MCP-1 gene expression was more strongly decreased by SPT than by GGT. 2. ICAM-1 and VCAM-1 gene expressions were more strongly decreased by SPT than by GGT 3. GGT significantly increased eNOS gene expression, but SPT did not. Conclusions: These findings suggest that GGT and SPT regulate gene expression related to anti-inflammatory effects in HUVECs. Clinical application of these Korean medicines to diseases related to dyslipidemia, such as cardiovascular disease, will require additional in vivo experiments to verify the anti-inflammatory effects of GGT and SPT.

Cloning of Mouse AQP-CD Gene

  • Jung, Jin-Sup;Kim, Joo-In;Oh, Sae-Ok;Park, Mi-Young;Bae, Hae-Rhan;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.2
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    • pp.195-200
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    • 1997
  • Water transport in highly-permeable membranes is facilitated by some specialized pathways, which are called aquaporins (AQP). AQP1 (AQP-CHIP) is the first recognized aquaporin identified from red cells and renal proximal tubules. Up until now 4 other aquaporin homologs have been reported. Each aquaporin has its unique tissue distribution and regulatory mechanims. To elucidate molecular mechanisms for their transcription regulation and tissue-specific expression isolation of aquaporin genes is required. To clone promoters of the AQP family mouse genomic library was screened by the 1st exon-specific probe of AQP4, and 5 different plaques were positively hybridized. Phage DNAs were purified and characterized by restriction mapping and sequencing. One of them is the mouse AQP-CD gene. The gene was consisted of 4 exons and the exon-intron boundaries of mouse AQP-CD gene were identified at identical positions in other related genes. The 5'-flanking region of AQP-CD gene contains one classic TATA box, a GATA consensus sequence, an E-box and a cyclic AMP-responsive element. The cloning of the mouse AQP-CD gene, of which product is expressed in the collecting duct and is responsible for antidiuresis by vasopressin, will contribute to understand the molecular mechanisms of tissue-specific expression and regulation of AQP-CD gene under various conditions.

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Lack of Effects of Peroxisome Proliferator-Activated Receptor Gamma Genetic Polymorphisms on Breast Cancer Risk: a Case-Control Study and Pooled Analysis

  • Park, Boyoung;Shin, Aesun;Kim, Kyee-Zu;Lee, Yeon-Su;Hwang, Jung-Ah;Kim, Yeonju;Sung, Joohon;Yoo, Keun-Young;Lee, Eun-Sook
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9093-9099
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    • 2014
  • A growing body of evidence suggests that the peroxisome proliferator-activated receptor-gamma ($PPAR{\gamma}$) gene may harbor targets for the chemoprevention of breast cancer. However, it is unclear whether polymorphisms in the $PPAR{\gamma}$ gene are associated with the susceptibility of breast cancer. We performed a candidate gene association study between $PPAR{\gamma}$ polymorphisms and breast cancer and a meta-analysis on the association of breast cancer with selected $PPAR{\gamma}$ variants. Six single nucleotide polymorphisms (SNPs) in the $PPAR{\gamma}$ gene were analyzed among 456 breast cancer patients and 461 controls from the National Cancer Center in Korea. Association between the polymorphisms and breast cancer risk were assessed using the Cochrane-Armitage test for trend and a multivariate logistic regression model. Two SNPs, rs3856806 and rs1801282, had been previously analyzed, thus enabling us to perform pooled analyses on their associations with breast cancer susceptibility. Our findings from the candidate gene association study showed no association between the $PPAR{\gamma}$ gene polymorphisms and breast cancer risk. A meta-analysis combining existing studies and our current study also refuted an association of the $PPAR{\gamma}$ gene with breast cancer. Our findings suggest that the $PPAR{\gamma}$ gene may not harbor variants that alter breast cancer susceptibility, although a moderate sample size might have precluded a decisive conclusion.

Association between Serotonin 2A(T102C) and 1B(G861C) Receptor Gene Polymorphism and Suicidal Attempt with Drug Intoxication in Korean Populations (음독 자살 시도와 세로토닌 수용체 2A(T102C) 및 1B(G861C) 유전자 다형성에 관한 연합연구)

  • Kim, Dong-Hyeon;Kwon, Young-Joon;Kim, Jae-Woo;Shim, Se-Hoon;Jung, Hee-Yeon
    • Korean Journal of Biological Psychiatry
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    • v.11 no.2
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    • pp.110-116
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    • 2004
  • Objectives:Recently, polymorphisms of several serotonin genes have been suggested to be associated with suicide, but the results are still unclear. We examined whether the T102C polymorphisms of the serotonin 2A receptor gene and the G861C polymorphisms of the serotonin 1B receptor gene were associated with suicidal behavior using drug intoxication. Methods:The subjects were 52 patients who visited emergency room with suicidal behaviors. Fifty controls were selected from healthy volunteers matched for sex and age to the suicide subjects. The polymorphisms were analyzed with TaqMan$^{(R)}$ assay using primers based on previous studies. Results:The T102C polymorphism of the serotonin 2A receptor gene showed no significant difference between the suicidal attempters and controls in both genotype and allele frequency analyses(p=0.179 and p=0.422, respectively). There was no statistically significant difference between the suicidal attempters and the controls in the G861C polymorphism of the serotonin 1B receptor gene and any significant effect of the genotype distributions or the allele frequencies was not observed(p=0.092 and p=0.987, respectively). Conclusion:These findings suggest that the T102C polymorphism in serotonin 2A receptor gene and the G861C polymorphism in serotonin 1B receptor gene are not related to the susceptibility to suicide attempts using drugs. To clarify the genetic influences of the serotonergic system on suicidal behavior, the polymorphisms of other candidate genes in the serotonergic system should be studied with larger numbers of subjects.

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The Effect of Gamidohongsamul-tang (GDT) on the Gene Expression Levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC Cells (가미도홍사물탕(加味桃紅四物湯)이 HUVEC cell 내에 eNOS, KLF2, ICAM-1, VCAM-1의 유전자 발현양에 대해 미치는 영향)

  • Lim, Hyun-chan;Jeon, Sang-yoon
    • The Journal of Internal Korean Medicine
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    • v.39 no.1
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    • pp.1-8
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    • 2018
  • Objectives: This study seeks to investigate the effects of Gamidohongsamul-tang (GDT) on the Gene expression levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC cells. Methods: HUVEC cells were treated at a concentration of 1, 10, 100 (${\mu}g/ml$) of Gamidohongsamul-tang (GDT). To measure the NOS, KLF2, ICAM-1 and VCAM-1 gene expression in HUVEC cells, the synthesized cDNA was subjected to polymerase chain reaction (PCR) and electrophoresis was performed to verify gene expression level. Results: 1. GDT significantly increased eNOS and KLF2 gene expression. 2. GDT significantly reduced ICAM-1 and VCAM-1 gene expression. Conclusions: These experiments suggest that Gamidohongsamul-tang (GDT) regulates gene expression related with anti-dyslipidemic effects in HUVEC cells. In order to clinically apply this to diseases related to dyslipidemia, such as cardiovascular disease, additional in vivo experiments are needed to verify the anti-dyslipidemic effects of GDT.

Development of the Mammalian Expression Vector System that can be Induced by IPTG and/or Lactose

  • Myung, Seung-Hyun;Park, Junghee;Han, Ji-Hye;Kim, Tae-Hyoung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1124-1131
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    • 2020
  • Techniques used for the regulation of gene expression facilitate studies of gene function and treatment of diseases via gene therapy. Many tools have been developed for the regulation of gene expression in mammalian cells. The Lac operon system induced with isopropyl β-D-1-thiogalactopyranoside (IPTG) is one of the employed inducible systems. IPTG mimics the molecular structure of allolactose and has a strong affinity for the corresponding repressor. IPTG is known to rapidly penetrate into mammalian cells and exhibits low toxicity. In the present study, we developed a new inducible expression system that could regulate the expression of genes in mammalian cells using IPTG. Here we confirm that unlike other vector systems based on the Lac operon, this expression system allows regulation of gene expression with lactose in the mammalian cells upon transfection. The co-treatment with IPTG and lactose could improve the regulatory efficiency of the specific target gene expression. The regulation of gene expression with lactose has several benefits. Lactose is safe in humans as compared to other chemical substances and is easily available, making this technique very cost-effective.

Effects of Woohwangcheongsim-won on Gene Expression in a Hypoxic Model of Cultured Rat Cortical Cells (배양한 흰쥐 대뇌세포의 저산소증 모델에서 우황청심원이 유전자 표현에 미치는 영향)

  • Park Dong-Wan;Kim Wan-Sik;Bae Cheol-hwan;Jeong Sung-Hyun;Shin Gil-Cho;Lee Won-Chul
    • The Journal of Korean Medicine
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    • v.25 no.3
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    • pp.123-136
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    • 2004
  • Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won (WC) on the in vitro neuronal development and alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E/sub 18/ rat cortical cells were grown in a neurobasal medium containing B27 supplement and various concentration of WC. Initial development of growth cone was investigated by phase-contrast microscopy, while dendritic spine formation and synaptogenesis were investigated by immunocytochemistry with SynGAPα(a postsynaptic marker) and synaptophysin (presynaptic marker) antibodies. Alteration in gene expression was analyses by microarray using rat 5K-TwinChips. Results : WC suppressed the development of growth cones and WC increased the number of dendritic spines at 20 and 50㎍/mL concentration but there was no statistical significance. Instead, it significantly decreased the number at 100㎍/mL. The expression of anti-apoptosis gene Bcl2-like 1 (Bcl211) increased (Global M=0.46), while Akt1 decreased. Proapoptosis genes Bad and PDCD2 increased. The expression of hemoglobin alpha 1 (probably neuroglobin) increased (Global M=0.93). The expression of antioxidants such as catalase, heme oxygenase (HO), and PRKAG2 gene increased. The expression PKC gene increased. The expression of retinoic acid receptor alpha (RARα) increased significantly (Global M=1.0). Conclusions : These data suggest that WC trends to suppress cellular activity slightly in normoxia and increases the expression of apoptosis-, antioxidation-, oxygen capture-related genes in hypoxia, but increases Bcl111 that anti-apoptosis gene, on the other hand increases Bad, PDCD2 that pro-apoptosis genes, too..

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RECK Gene Promoter rs10814325 Polymorphism in Egyptian Patients with Hepatocellular Carcinoma on Top of Chronic Hepatitis C Viral Infection

  • Fakhry, Amal Bahgat;Ahmed, Asmaa Ismail;AbdelAlim, Mahmoud Abdo;Ramadan, Dalia Ibrahim
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2383-2388
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    • 2016
  • Background: The reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) gene is a novel transformation suppressor gene linked to several malignancies. Objective: To analyze any association between RECK gene rs10814325 single-nucleotide polymorphism (SNP) and HCC susceptibility with various clinicopathological and laboratory data. Materials and Methods: RECK gene rs10814325 SNP was estimated, using real-time PCR, in 30 HCC patients on top of HCV infection, 30 HCV related cirrhotic patients and 30 healthy controls. Results: No special pattern of association could be detected on comparing the RECK gene rs10814325 genotypes(P=0.5), or alleles(P=0.49) among the studied groups. HCC patients with TT genotype had younger age (mean of $54.1{\pm}6.0$ years vs $60.6{\pm}10.2$ years for TC/CC genotypes, P=0.035). Abdominal distension was significantly greater in TT genotype patients (75% vs 30%for TC/CC genotypes, P=0.045). The TT genotype was present in 75% of patients with lymph node metastasis. Serum GGT levels were higher in TT genotype patients [80 (48.5-134.8) IU/L vs 40 (33-87.5) IU/L for TC/CCgenotypes], and lower limb edema was observed in 60% for TT vs 20% for TC/CCgenotypes, but both just failed to reach significance (p=0.05 and p=0.06 respectively). Conclusions: RECK gene rs10814325 T>C could not be considered a risk factor for HCC development on top of HCV, but may be related to the disease progression and metastasis.

Antitumor Effect of an Adenoviral Cytosine Deaminase/Thymidine Kinase Fusion Gene in C6 Glioma Cells (아데노 바이러스 Cytosine Deaminase/Thymidine Kinase 융합 유전자의 항 종양효과)

  • Kim, Young Woo;Choi, Jae Young;Chang, Jin Woo;Park, Yong Gou;Chung, Sang Sup
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup1
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    • pp.13-19
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    • 2001
  • Objective : We investigated the feasibility of a double suicide gene/prodrug therapy, involving direct introduction of the herpes simplex virus Type 1 thymidine kinase(TK) gene and the Escherichia coli cytosine deaminase(CD) gene, via a recombinant adenoviral vector and ganciclovir(GCV) and/or 5-fluorocytosine(5-FC) treatment, in C6 glioma cells. Methods : Efficient gene transfer and transduction of C6 glioma cells via a recombinant adenovirus were evaluated by infecting cells with adenovirus bearing the ${\beta}$-galactosidase gene and then staining cells with 5-bromo-4-chloro-3-indolyl-13-D-galactoside. CD/TK expression in cells infected with adenovirus bearing the CD/TK gene(ad-CD/TK) was examined by immunoblotting analysis. For in vitro cytotoxicity experiments, the cells were infected with ad-CD/TK or ad-${\Delta}E1$(as a control). After addition of a variety of concentrations of GCV and 5-FU, either separately or in combination, cell viability was determined by staining the cells with crystal violet solution 6 days after infection. Result : C6 glioma cells were efficiently transduced with recombinant adenoviral vector at multiplicities of infection of 200 or more. In vitro cytotoxicity of GCV and/or 5-FC, either alone or in combination, was exclusively observed in the cells transduced with ad-CD/TK. Obvious cytotoxicity(>50% inhibition) was observed in the presence of 5-FC at concentrations greater than 30ug/ml or GCV at concentrations greater than 0.3ug/ml at a multiplicity of infection of 100. Additionally, cytotoxicity in the presence of both GCV and 5-FC was greater than that after sinlge-prodrug treatments, indicating additive effects of the prodrug treatments. Conclusion : The administration of a double-suicide gene/prodrug therapy might have great potential in the treatment of brain tumors.

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