• Title/Summary/Keyword: gene effect

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Effect of ganglioside GD3 synthase gene expression on VSMC proliferation via ERK1/2 pathway, cell cycle progression and MMP-9 expression

  • Lee, Young-Choon;Kim, Cheorl-Ho
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.81-90
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    • 2004
  • Sialic acid containing glycosphingolipids (gangliosides) have been implicated in the regulation of various biological phenomena such as atherosclerosis. Recent report suggeststhat exogenously supplied disialoganglioside (GD3) serves a dual role in vascular smooth muscle cells (VSMC) proliferation and apoptosis. However, the role of the GD3 synthase gene in VSMC responses has not yet been elucidated. To determine whether a ganglioside is able to modulate VSMC growth. the effect of overexpression of the GD3 synthase gene on DNA synthesis was examined. The results show that the overexpression of this gene has a potent inhibitory effect on DNA synthesis and ERK phosphorylation in cultured VSMC in the presence of PDGF. The suppression of the GD3 synthase gene was correlated with the down-regulation of cyclinE/CDK2. the up-regulation of the CDK inhibitor p21 and blocking of the p27 inhibition,whereas up-regulation of p53 as the result of GD3 synthase gene expression was not observed. Consistently, blockade of GD3 function with anti-GD3 antibody reversed VSMC proliferation and cell cycle proteins. The expression of the CD3 synthase gene also led to the inhibition of TNF--induced matrix metalloproteinase-9 (MMP-9) expression in VSMC as determined by zymography and immunoblot. Furthermore, GD3 synthase gene expression strongly decreased MMP-9 promoteractivlty in response to TNF-. This inhibition was characterized by the down-regulation of MMP-9,which was Iranscriptionally regulated at NF-B and activation protein-1 (AP-1) sites in the MMP-9promoter Finally, the overexpression of MMP-9 in GD3 synthase transfectant cells rescued VSMC proliferation. However MMP-2 overexpression was not affected the cell proliferation. These findings suggest that the fl13 synthase gene represents a physiological modulator of VSMC responses that may contribute to plaque instability in atherosclerosis.

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The Effect of $17{\beta}-Estradiol$ on the Gene Expression of IGF-I and Bone Matrix Protein in the Osteoblast-Like Cell (골아세포의 IGF-I 유전자 발현 및 골기질 단백질에 대한 $17{\beta}-estradiol$의 영향)

  • Yang, Won-Suk;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.375-390
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    • 2000
  • The purpose of this study is to evaluate the expression ofIGF-I, considered as the mediator of action of estrogen, and IGF-IA and IGF-IB, alternative slicing form of IGF-I, using $17{\beta}-estradiol$ in MC3T3-E1 cells. We observed the effect on type I collagen and osteopontin gene expression and DNA synthetic activity of MC3T3-E1 cells, added by estrogen, IGF-I and combination and the interactionon proliferation and differentiation of MC3T3-E1 cells. The results were as follows :RT-PCR experiment for observing timedependantIGF-I gene expression patternshowed IGF-IA and IB gene expression in both of control and test group. In these IGF-IA gene expression was appeared predominantly. In control, IGF-I geneexpression level was maintained until 24hr and then decreased gradually. In testgroup, IGF-I gene expression level increased as time goes by. Experiment measuring DNA synthetic activity, as it is added by $17{\beta}-estradiol$, IGF-I and combination, showed that first day , there was the tendency of more increase of synthetic activity in all test group than control but no statical significance(P>0.05), and third day, there was more increase of DNA synthetic activity in $17{\beta}-estradiol$ group and combination group and it was statically significant. (P<0.005) Experiment for observing type I collagen gene expression pattern showed more increase of expression in $17{\beta}-estradiol$ group than control and no significant difference in IGF-I group and combination group. Experiment for observing osteopontin gene expression pattern showed no significant difference in control and test group. In conclusion, $17{\beta}-estradiol$ in MC3T3- E1 cells increased IGF-I gene and DNA synthetic activity simultaneously, therefore it appeared that IGF-I is related to the action of estrogen. Combination treatment of IGF-I and $17{\beta}-estradiol$ has effect on cell proliferation but this effect is lower than IGF-I or $17{\beta}-estradiol$ alone. However, combination treatment has not great effect on type I collagen or osteopontin gene expression thus little effect of cell differentiation.

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Effect of Corydalis Tuber on the inhibition of proliferation of human uterine leiomyoma cell and apoptotic gene expression (현호색(玄胡索)이 자궁근종세포의 증식 억제와 Apoptosis 관련 유전자 발현에 미치는 영향)

  • Lee, Hee-Jae;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.214-225
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    • 2006
  • Purpose : This study was aimed to investigate the inhibitory effect of Corydalis Tuber on the proliferation of human uterine leiomyoma cell and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of suvival cells treated with indicated concentration of Corydalis Tuber and investigated cell viability by MTS assay. Furthermore, flow cytometric analyis were used to dissect between necrosis and apoptosis related with cell cycle and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the proliferation of uterine leiomyoma cell treated with Corydalis Tuber was increased in a concentration and time proportional. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was not investigated in uterine leiomyoma cell treated Corydalis Tuber but showed G2/M phase prolongation. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing concentration, but p53 was not exchanged. 4) The dephosphorylation of pRb gene were increased dependent on treatment concentration and pro-caspase 3, CDK4 were not exchanged. Conclusion : This study showed that Corydalis Tuber have the inhibitory effect on the proliferation of human uterine leiomyoma cell but the effect was thoughted no relationship with apoptosis. The inhibitory effect was suggested that dephosphorylation of pRb gene induced with increasing p21, p27 prolonged cell division in G2/M phase.

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Gene Expression of Heart and Adipocyte Fatty Acid-binding Protein in Chickens by FQ-RT-PCR

  • Tu, Yunjie;Su, Yijun;Wang, Kehua;Zhang, Xueyu;Tong, Haibing;Gao, Yushi
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.8
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    • pp.987-992
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    • 2010
  • This study was to detect the expression of heart fatty acid-binding protein (H-FABP) and adipocyte fatty acid-binding protein (A-FABP) gene mRNA in different tissues of Rugao and Luyuan chickens at 56 d and 120 d by real-time fluorescence quantitative reverse transcription polymerase-chain reaction (FQ-RT-PCR). The primers were designed according to the sequences of HFABP, A-FABP and GAPDH genes in Gallus gallus, which were used as target genes and internal reference gene, respectively. The levels of H-FABP and A-FABP gene expression were detected by SYBR Green I FQ-RT-PCR. The relative H-FABP and A-FABP gene mRNA expression level was calculated with 2-$^{{\Delta}Ct}$. Melting curve analysis showed a single peak of three genes. Intramuscular fat (IMF) content in breast muscle and leg muscle of the two chicken breeds at 120 d was higher than at 56 d. IMF content in breast muscle and leg muscle at 56 d and 120 d in Luyuan was significantly higher than in Rugao, however, abdominal fat of Luyuan was significantly lower than that of Rugao. The relative H-FABP gene mRNA expression level in cardiac muscle was the highest in both chicken breeds. The relative H-FABP and A-FABP gene expression of different tissues in Luyuan was higher than in Rugao. H-FABP gene mRNA expression had a negative effect on IMF of leg and breast muscles, and was significantly negatively correlated with IMF content. The relative A-FABP gene mRNA level in abdominal fat was higher than in liver. The A-FABP gene mRNA was not expressed in leg, breast and cardiac muscles. A-FABP gene mRNA expression level was significantly positively correlated with abdominal fat and had a significant effect on abdominal fat but not IMF content.

Effect of Single Nucleotide Polymorphisms of Acetyl-CoA Carboxylase α(ACACA) Gene on Carcass Traits in Hanwoo (Korean Cattle)

  • Shin, Sung-Chul;Heo, Jae-Pil;Chung, Eui-Ryong
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.6
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    • pp.744-751
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    • 2011
  • Meat production and quality traits in beef cattle are largely affected by genetic factors. Acetyl-Coenzyme A carboxylase-${\alpha}$ (ACACA) plays a key role in the regulation and metabolism of fatty acid biosynthesis in mammalian animals. The gene encoding ACACA enzyme was chosen as a candidate gene for carcass and meat traits. In this study, we investigated effects of single nucleotide polymorphisms (SNPs) in the ACACA gene on beef carcass and meat traits in Hanwoo (Korean cattle) populations. We have sequenced a fragment of intron I region of the Hanwoo ACACA gene and identified two SNPs. Genotyping of the two SNP markers (g.2344T>C and g.2447C>A) was carried out using PCR-SSCP analysis in 309 Hanwoo steers to evaluate their association with carcass and meat production traits. The g.2344C SNP marker showed a significant increasing effect on LW (p = 0.009) and CW (p = 0.017). Animals with the CC genotype had higher CW and LW compared with TT and TC genotypes (p<0.05). The g.2447A SNP marker was associated with higher MC (p = 0.019). Animals with the AA genotype had higher MC than animals with CC and CA genotypes (p<0.05). Although the degree of linkage disequilibrium (LD) was not strong between g.2344T>C and g.2447C>A in the LD analysis, four major haplotype classes were formed with two SNP information within the ACACA gene. We constructed haplotypes using the HaploView software package program and analyzed association between haplotypes and carcass traits. The haplotype of ACACA gene significantly affected the LW (p = 0.027), CW (p = 0.041) and MC (p = 0.036). The effect of h1 haplotype on LW and CW was larger than that of h3 haplotype. Animals with the h1 haplotype also had greater MC than did animals with h2 haplotype. Consequently, the ACACA gene could be useful as a DNA marker for meat production traits such as carcass yield and meat contents in Hanwoo.

Gadobutrol-dendrimer effects on metastatic and apoptotic gene expression

  • Kebriaezadeh, Abbas;Ashrafi, Sepehr;Rasouli, Rahimeh;Ebrahimi, Seyed Esmaeil Sadat;Hamedani, Morteza Pirali;Assadi, Artin;Saffari, Mostafa;Ardestani, Mehdi Shafiee
    • Advances in nano research
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    • v.4 no.2
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    • pp.145-156
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    • 2016
  • Dendrimers are one of the most appropriate nanocaries for imaging moieties in imaging applications.The purpose of this study was the evalution of cytotoxicity and inducing apoptosis of dendrimers. This study was conducted in order to investigate the metastasis suppression effect of dendrimer in human breast MCF-7 cell line and finding the nanoparticle protein corona in biological enviromental. Dendrimer cytotoxicity effect was assessed by MTT assay. The mRNA experession level of KAI1 as a metastasis suppressor gene, Bax as Pro- apoptotic gene, Bcl-2 as an anti-apoptotic gene and GAPDH as a housekepping gene were determined by real-time PCR assays.concentration-dependent nanoparticle cytotoxicity effect was proofed at range of 1-2 mg/mL in 24 hours, significant upregulation of mRNA expression of Bax, was observed whereas expression of anti-apoptotic Bcl-2 was down-regulated, also expression of metastasis suppressor gene KAI1 was up-regulated. So far a few studies confirmed apoptosis enhancement effect of dendrimers in MCF-7 cell line via bax/bcl-2 pathways. dendrimer nanoparticles was able to act as metastase inhibitor via upregulation of KAI1 gene.

Genetic Analysis for Agronomic, Chemical , and Leaf Chararters According to Stalk Position in Flue-Cured Tobacco (Nicotiana tabacum L. ) II. Analysis of Gene Action by Diallel Crosses (황색종 연초(Nicotiana tabacum L.)의 주요형질과 엽위별 엽형질에 대한 유전분석 II. 이면교배에 의한 유전자 작용분석)

  • 조수헌
    • Journal of the Korean Society of Tobacco Science
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    • v.9 no.2
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    • pp.37-47
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    • 1987
  • This experiment was conducted to obtain basic information on breeding of flue-cured tabacco varieties . Nine cultivars and partial diallel set of 36 Fl hybrids were grown at Daegu Experiment Station, Korea Ginseng & Tobacco Research Institute in 1983. Partial dominance with high additive gene effect was observed for yield, leaves per plant , days to flower, leaf weight, nicotine and reducing sugar content. partial dominance with additive and dominant gene effect was observed for stalk height , leaf width midrib weight Overdominance with high dominant gene effect was observed for leaf length and midrib width. The directions of dominance were positive for yield, stalk height, leaf weight, leaf length, leaf width, midrib weight, midrib width and reducing sugar content, and the negative was days to flower. The estimates of effective genes were 1 for leaves per plant, 2 for stalk height, days to flower and leaf shape. 3 to 6 for leaf length, leaf width, leaf weight and midrib weight.

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Effect of Dexamethasone Preincubation on Polymer-Mediated Gene Delivery

  • Choi, Joon-Sig;Lee, Min-Hyung
    • Bulletin of the Korean Chemical Society
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    • v.26 no.8
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    • pp.1209-1213
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    • 2005
  • Nuclear membrane is one of the main barriers in intracellular delivery of genetic materials. The previous report showed that glucocorticoid receptor dilated the nuclear pore to 60 nm in the presence of a ligand. It was also suggested that the transport of genetic material to nucleus might be facilitated by glucocorticoid. In this study, the effect of glucocorticoid preincubation in the polymeric gene delivery was investigated. The cells were preincubated with dexamethasone, a potent glucocorticoid, and transfection assays were performed with polyethylenimine (PEI) and polyamidoamine (PAMAM) dendrimer. As a result, the transfection efficiency of PEI or PAMAM to the cells in the presence of dexamethasone was enhanced, compared to the cells without dexamethasone. This effect was not observed in the cells preincubated with cholesterol. The polymer/DNA complex was stable in the presence of dexamethasone. In addition, the cytotoxicities of the polymeric carriers to the cells were observed in the presence of dexamethasone. In conclusion, dexamethasone enhances the transfection efficiency of polymeric carriers and may be useful in the development of polymeric gene carriers.

Developmental Gene Expression of Antimicrobial Peptide PR-39 and Effect of Zinc Oxide on Gene Regulation of PR-39 in Piglets

  • Wang, Y.Z.;Xu, Z.R.;Lin, W.X.;Huang, H.Q.;Wang, Z.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1635-1640
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    • 2004
  • Two experiments were conducted to evaluate developmental gene expression of antimicrobial peptide PR-39 and effect of zinc oxide on gene regulation of PR-39 in piglets using semi-quantitative RT-PCR analysis. In experiment 1, fifteen female Tai-Hu pigs (a local breed in China) in five groups, each of three pigs at 1, 14, 28, 42 and 56 days of age were used to determine effect of age and weaning on mRNA expression of PR-39. In experiment 2, nine groups of pigs (total seventy-two female 36 days-age weanling Tai-Hu piglets) were assigned to three treatments (${ZnO}_0$, ${ZnO}_{100}$ and ${ZnO}_{3000}$). The feeding experimental period lasted 15 days. After feeding experiment, nine pigs with three animals in each treatment were chosen to determine the effect of ZnO on PR-39 mRNA expression of pigs. The results showed that PR-39 mRNA levels increased steadily in postnatal day 1-28 (preweaning), and weaning significantly decreased PR-39 mRNA expression of piglets (p<0.05). ${ZnO}_{3000}$ (3,000 mg zinc/kg diet) significantly increased PR-39 mRNA expression (p<0.05) when piglets were feed ${ZnO}_{3000}$ diet for 15 days. ${ZnO}_{100}$ (100 mg zinc/kg diet) also increased PR-39 gene expression, but the result was not statistically significant (p>0.05). The result was in accordance with the effect of ${ZnO}_{3000}$ and ${ZnO}_{100}$ on weight gain of piglets and prevention of diarrhea.

Effect of Chestnut Bark Extracts on Tyrosinase Gene Expression (율피 추출물이 티로시나아제 유전자 발현에 미치는 효과)

  • Chin Jong-Eon;Kim Kwan-Chun
    • Journal of environmental and Sanitary engineering
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    • v.20 no.3 s.57
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    • pp.10-16
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    • 2005
  • Chestnut bark extract by methanol repressed the expression of tyrosinase gene of B16 mouse melanoma cell containing tyrosinase promoter. $10{\mu}g/ml{\ell}\;100{\mu}g/m{\ell}$, $1mg/m{\ell}$ of the extract repressed expression of tyrosinase gene about $38\%,\;47\%,\;and\;78\%$, respectively, compared with control. In the MTT assay, the same extract exhibited very low cytotoxicity at $1{\mu}g/m{\ell},\;10{\mu}g/m{\ell},\;100{\mu}g/m{\ell},\;and\;1mg/m{\ell}$, respectively. The fractions of Methylene chloride and ethyl acetate did not showed the repressive effect on the expression of tyrosinase gene, but the fraction of butyl alcohol repressed highly at $10{\mu}g/m{\ell}\;and\;100{\mu}g/m{\ell}$.