• Title, Summary, Keyword: gene experession

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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.

Selection of Constitutive Promoter for Exoinulinase Production in Fed-Batch Culture of Recombinant Yeast (재조합 효모의 유가배양에서 Exoinulinase생산을 위한 Promoter의 선별)

  • 김이경;고지현;김연희;김성구;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.206-211
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    • 2001
  • In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

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Sex Steroids Regulate Expression of Genes Containing Long Interspersed Elements-1s in Breast Cancer Cells

  • Chaiwongwatanakul, Saichon;Yanatatsaneejit, Pattamawadee;Tongsima, Sissades;Mutirangura, Apiwat;Boonyaratanakornkit, Viroj
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.4003-4007
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    • 2016
  • Long interspersed elements-1s (LINE-1s) are dispersed all over the human genome. There is evidence that hypomethylation of LINE-1s and levels of sex steroids regulate gene expression leading to cancer development. Here, we compared mRNA levels of genes containing an intragenic LINE-1 in breast cancer cells treated with various sex steroids from Gene Expression Omnibus (GEO), with the gene expression database using chi-square analysis (http://www.ncbi.nlm.nih.gov/geo). We evaluated whether sex steroids influence expression of genes containing an intragenic LINE-1. Three sex steroids at various concentrations, 1 and 10 nM estradiol (E2), 10 nM progesterone (PG) and 10 nM androgen (AN), were assessed. In breast cancer cells treated with 1 or 10 nM E2, a significant percentage of genes containing an intragenic LINE-1 were down-regulated. A highly significant percentage of E2-regulated genes containing an intragenic LINE-1 was down-regulated in cells treated with 1 nM E2 for 3 hours (p<3.70E-25; OR=1.91; 95% CI=2.16-1.69). Similarly, high percentages of PG or AN-regulated genes containing an intragenic LINE-1 wwere also down-regulated in cells treated with 10 nM PG or 10 nM AN for 16 hr (p=9.53E-06; OR=1.65; 95% CI=2.06-1.32 and p=3.81E-14; OR=2.01; 95% CI=2.42-1.67). Interestingly, a significant percentage of AN-regulated genes containing an intragenic LINE-1 was up-regulated in cells treated with 10 nM AN for 16 hr (p=4.03E-02; OR=1.40; 95% CI=1.95-1.01). These findings suggest that intragenic LINE-1s may play roles in sex steroid mediated gene expression in breast cancer cells, which could have significant implications for the development and progression of sex steroid-dependent cancers.

The Analysis of the Cytokine Expression in Musculoskeletal Tumors (근골격계 종양에서 사이토카인 발현의 분석)

  • Lee, Joon-Han;Kwak, Eun-Seok;Shon, Oog-Jin;Kim, Hee-Sun;Shin, Duk-Seop
    • Yeungnam University Journal of Medicine
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    • v.20 no.2
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    • pp.187-196
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    • 2003
  • The cytokines are the hormone-like proteins, which are produced in the mononuclear cells. They have many roles, such as immune mediators, cell differentiations, angiogenesis. The chemokines have chemotactic effects which control the host immune response. There were few reports about the cytokines associated with musculoskeletal tumors. From late 1980s, the cytokine studies of bone tumors such as osteosarcoma were started, but most studies for benign and malignant musculoskeletal tumors were left to be explored. To evaluate the characteristics of the cytokines in variable musculoskeletal tumors, tissues were obtained from the seven patients who visited the Yeungnam University hospital from February to July 2000. They were lipoma (1 case), parosteal osteoma (1 case), enchondroma (2 cases), pigmented villonodular synovitis (1 case), ganglion (1 case), and metastaic squamous cell carcinoma (1 case). The gene experession of the cytokines were analyzed by RNase protection assay (RPA) and reverse transcription-polymerase chain reaction (RT-PCR). The lipoma and parosteal osteoma expressed MIP-$1{\beta}$, and IP-10 genes. The two enchondromas showed different results, one expressed all of MIP-$1{\alpha}$, MIP-$1{\beta}$ and IP-10 genes but the other expressed none of above. The pigmented villonodular synovitis strongly expressed MIP-$1{\alpha}$ and IP-10 when compared with the other cases. The ganglion did not express all of the chemokines mentioned above. And the metastatic squamous cell carcinoma expressed all of the chemokines and especially IP-10 was highly expressed. Even though this study has only a few cases, these results provide a basis for the cytokine mediating network study in musculoskeletal tumors.

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Estrogenic Potentials of Organochlorine Pesticides which Detected in Korean Adipose Tissue (한국인의 지방조직에서 검출되는 유기염소제 농약의 에스트로겐 활성)

  • Oh Seung-Min;Kim Mi-Young;Lee Hee-Sung;Lee Do-Han;Lee Sang-Ki;Chung Kyu-Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.19 no.2
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    • pp.207-216
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    • 2004
  • 유기염소계 농약은 화학적으로 안정하고, 지용성이 크며 체내 대사에 대해 저항성을 가지고 있어서 인체 및 생태계의 생물체 중 축적성이 매우 크다. 또한 대부분 내분비계 장애물질로 분류되어 있어 관심의 대상이 되고 있다. 유기염소계 농약은 잔류성으로 인하여 사용이 금지되었음에도 불구하고 인체에서 검출되고 있으며 한국인의 지방조직을 분석한 결과 9종의 유기염소계 농약이 주로 축적되어 있음이 보고된 바 있다. 본 연구에서는 이들 유기염소계 농약의 에스트로겐 활성을 MCF-7 BUS cell을 이용한 E-screen assay competitive binding assay 및 pS$_2$ gene experession assay에 의해 조사하였다. o,p'-DDT,p, p'-DDT,p,p'-DDD,p,p'-DDE등 4종의 유기염소계 농약은 에스트로겐 수용체에 대한 ligand의존적인 작용기전에 의해 에스트로겐 활성을 가지며, $\alpha$-, $\beta$-, ${\gamma}$-, $\delta$-BHC, dieldrin등 5종의 유기염소계 농약은 lingand-비의존적 작용기전에 의해 에스트로겐 활성을 보였다. 또한 이들 유기염소계 농약을 혼합 투여하여 에스트로겐 활성을 관찰한 결과 DDT류의 경우에는 단독투여시 보다 그 대사체와 혼합 투여할 때 에스트로겐 활성에 상승적 효과가 나타났으며 o,p'-와 p,p'-DDT의 두 이성질체를 혼합 투여할 경우가 단독 투여시 보다 상승적 효과가 나타났다. 따라서 지방조직에서 검출되는 유기염소계농약은 상호작용에 의해 개별 물질이 나타내는 내분비계장애작용 보다 실제로는 강한 효과를 나타낼 것으로 추정되었다.