• Title/Summary/Keyword: KIT gene

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Strong concordance between RNA structural and single nucleotide variants identified via next generation sequencing techniques in primary pediatric leukemia and patient-derived xenograft samples

  • Barwe, Sonali P.;Gopalakrisnapillai, Anilkumar;Mahajan, Nitin;Druley, Todd E.;Kolb, E. Anders;Crowgey, Erin L.
    • Genomics & Informatics
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    • v.18 no.1
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    • pp.6.1-6.9
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    • 2020
  • Acute leukemia represents the most common pediatric malignancy comprising diverse subtypes with varying prognosis and treatment outcomes. New and targeted treatment options are warranted for this disease. Patient-derived xenograft (PDX) models are increasingly being used for preclinical testing of novel treatment modalities. A novel approach involving targeted error-corrected RNA sequencing using ArcherDX HemeV2 kit was employed to compare 25 primary pediatric acute leukemia samples and their corresponding PDX samples. A comparison of the primary samples and PDX samples revealed a high concordance between single nucleotide variants and gene fusions whereas other complex structural variants were not as consistent. The presence of gene fusions representing the major driver mutations at similar allelic frequencies in PDX samples compared to primary samples and over multiple passages confirms the utility of PDX models for preclinical drug testing. Characterization and tracking of these novel cryptic fusions and exonal variants in PDX models is critical in assessing response to potential new therapies.

Immunomodulatory effects of fermented Platycodon grandiflorum extract through NF-κB signaling in RAW 264.7 cells

  • Park, Eun-Jung;Lee, Hae-Jeung
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.453-462
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    • 2020
  • BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

Effects of Juakium-derivative on Experimental Aplastic Anemia (좌귀음가미방이 실험적 재생불량성빈혈에 미치는 영향)

  • 박신명;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.23 no.1
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    • pp.133-144
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    • 2002
  • Objectives : Korean traditional medicinal herbs have been used to improve the function of the hematopoietic system. The purpose of this study is to examine the hematopoietic effects of Juakium-derivative on aplastic anemia. Methods : In vitro, after bone marrow cells of mice with aplastic anemia induced by benzene were cultured with Juakium-derivative, we measured the gene expressions of hematopoietic cytokines, the intracellular TPO and SCF expression and the colony number of CFU-GEMM and BFU-E. Results : The results are summarized as follows : 1. The gene expressions of hematopoietic cytokines (TPO, c-mpl, SCF, c-kit, IL-3, EPO, EpoR, GM-CSF) and the TPO and SCF expression of the group treated with Juakium-derivative increased significantly more than those of the control group. 2. The colony number of CFU-GEMM and BFU-E in the rEPO plus rIL-3 plus Juakium-derivative group increased significantly compared with the only rEPO plus rIL-3 group. Conclusion : It was acknowledged that Juakium-derivative has hemato-potentiating effects on aplastic anemia induced by benzene and it is expected that Juakium-derivative can be used clinically for the patient with hematopoietic system disorder. This study could also be used further as important research data for Korean oriental medicine about hemato-potentiating. The exact mechanism explaining our study would need to be studied further.

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MiRPI: Portable Software to Identify Conserved miRNAs, Targets and to Calculate Precursor Statistics

  • Vignesh, Dhandapani;Parameswari, Paul;Im, Su-Bin;Kim, Hae-Jin;Lim, Yong-Pyo
    • Genomics & Informatics
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    • v.9 no.1
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    • pp.39-43
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    • 2011
  • MicroRNAs (miRNAs) are recently discovered small RNA molecules usually resulting in translational repression and gene silencing. Despite the fact that specific cloning of small RNA's is a method in practice, computational identification of miRNA's has been a major focus recent days, since is a rapid process following AB initio and sequence alignment methods. Here we developed new software called MiRPI that aims to identify the highly conserved miRNAs without any mismatches from given fasta formatted gene sequences by using non-repeated miRNA dataset of the user's interest. The new window embedded with the software is used to identify the targets for inputted mature miRNAs in the mRNA sequences. Also MiRPI is designed to measure the precursor miRNA statistics, majorly focusing the Adjusted Minimum Folding free Energy (AMFE) and Minimum Folding free Energy Index (MFEI), the most important parameters in miRNA confirmation. MiRPI is developed by PERL (Practical Extraction and Report Language) and Tk (Tool kit widgets) scripting languages. It is user friendly, portable offline software that works in all windows OS, sized to 3 MB.

SETDB1 regulates SMAD7 expression for breast cancer metastasis

  • Ryu, Tae Young;Kim, Kwangho;Kim, Seon-Kyu;Oh, Jung-Hwa;Min, Jeong-Ki;Jung, Cho-Rok;Son, Mi-Young;Kim, Dae-Soo;Cho, Hyun-Soo
    • BMB Reports
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    • v.52 no.2
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    • pp.139-144
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    • 2019
  • Breast cancer (BRC) is the most invasive cancer in women. Although the survival rate of BRC is gradually increasing due to improved screening systems, development of novel therapeutic targets for inhibition of BRC proliferation, metastasis and recurrence have been constantly needed. Thus, in this study, we identified overexpression of SETDB1 (SET Domain Bifurcated 1), a histone methyltransferase, in RNA-seq data of BRC derived from TCGA portal. In Gene Ontology (GO) analysis, cell migration-related GO terms were enriched, and we confirmed down-regulation of cell migration/invasion and alteration of EMT /MET markers after knockdown of SETDB1. Moreover, gene network analysis showed that SMAD7 expression is regulated by SETDB1 levels, indicating that up-regulation of SMAD7 by SETDB1 knockdown inhibited BRC metastasis. Therefore, development of SETDB1 inhibitors and functional studies may help develop more effective clinical guidelines for BRC treatment.

Association of Cytochrome-17 (MspA1) Gene Polymorphism with Risk of Gall Bladder Stones and Cancer in North India

  • Dwivedi, Shipra;Agrawal, Sarita;Singh, Shraddha;Madeshiya, Amit Kumar;Singh, Devendra;Mahdi, Abbas Ali;Chandra, Abhjeet
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5557-5563
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    • 2015
  • Background: Cholelithiasis is associated in 54%-98% of patients with carcinoma of the gallbladder, and a high incidence among females suggests a role of female hormones in the etiology of the disease. Cytochrome $P450C17{\alpha}$ (CYP-17) is a key enzyme involved in estrogen metabolism and polymorphisms in CYP-17 are associated with altered serum levels of estrogens. Thus, we investigated whether the CYP-17 MspA1 gene polymorphism might impact on risk of gall bladder cancers or gallstones, as well as to determine if this gene polymorphism might be linked with estrogen serum levels and lipid profile among the North Indian gall bladder cancer or gallstone patients. Materials and Methods: CYP-17 gene polymorphisms (MspA1) were genotyped with PCR-RFLP in cancer patients (n=96), stone patients (n=102), cancer + stone patients (n=52) and age/sex matched control subjects (n= 256). Lipid profile was estimated using a commercial kit and serum estrogen was measured using ELISA. Results: The majority of the patients in all groups were females. The lipid profile and estrogen level were significantly higher among the study as compared to control groups. The frequency of mutant allele A2 of CYP17 MspA1 gene polymorphism was higher among cancer (OR=5.13, 95% CI+3.10-8.51, p=0.0001), stone (OR=5.69, 95%CI=3.46-9.37, p=0.0001) and cancer + stone (OR=3.54, 95%CI=1.90-6.60, p=0.0001) when compared with the control group. However there was no significant association between genotypes of CYP17 MspA1 gene polymorphism and circulating serum level of estrogen and lipid profile. Conclusions: A higher frequency of mutant genotype A1A2 as well as mutant allele A2 of CYP-17 gene polymorphism is significantly associated with risk of gallbladder cancer and stones. Elevated levels of estrogen and an altered lipid profile can be used as predictors ofgall bladder stones and cancer in post menopausal females in India.

INDUCTION OF APOPTOSIS IN ORAL CANCER CELL LINE THROUGH AN RECOMBINANT HCCS-1 ADENOVIRUS (재조합 HCCS-1 아데노바이러스를 이용한 구강암 세포주의 세포사멸 유발)

  • Kim, Chang-Hyen;Lee, Dong-Ju;Lee, ll-Kyu;Kim, Myung-Jin;Kim, Jin-Woo;Pyo, Sung-Woon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.4
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    • pp.306-311
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    • 2005
  • Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation. This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

Description of unrecorded wild yeasts from soil in Republic of Korea under cold conditions

  • Soohyun Maeng;Sathiyaraj Srinivasan
    • Journal of Species Research
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    • v.13 no.2
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    • pp.142-146
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    • 2024
  • The purpose of this study was to isolate and identify wild yeasts from soil collected in Daegu City and Cheongyang County, Republic of Korea. Among 11 strains isolated in this study, nine strains were previously reported and two strains were unreported in Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of the D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation test are done using API 20C AUX kit. All strains were assigned to the phylum Basidiomycota. Of the two unrecorded yeast strains, CY-9-10C belongs to the genus Mrakia (family Mrakiaceae, order Cystofilobasidiales, class Tremellomycetes) and PG3-4-10C belongs to the genus Slooffia (family Chrysozymaceae, order Microbotryomycetes incertae sedis, class Microbotryomycetes). Both strains had oval-shaped and polar budding cells. This research described the morphological and biochemical properties of the two unreported yeast species that had not officially reported in Korea.

Report of two unrecorded yeast species in the class Tremellomycetes

  • Seonjae Kim;Sathiyaraj Srinivasan
    • Journal of Species Research
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    • v.13 no.2
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    • pp.136-141
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    • 2024
  • The purpose of this study is to isolate and identify wild yeasts from the soil samples collected in Daegu and Daejeon City, Republic of Korea. Among 15 strains isolated in this study, 13 strains were previously reported and two strains had not been reported in Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation tests were done using API 20C AUX kit. All strains were assigned to the phylum Basidiomycota. The two unrecorded yeast strains, PG2-2-10C and DJ2-14-10C, belong to the genus Holtermanniella (family Holtermanniaceae, order Holtermanniales, class Tremellomycetes) and Goffeauzyma (family Filobasidiaceae, order Filobasidiales, class Tremellomycetes), respectively. The two unrecorded yeast strains had oval shape and polar budding cells. This research describers the morphological and biochemical properties of the two unreported yeast species that had not officially reported in Korea.

Isolation and characterization of two unrecorded yeast species in the order Filobasidiales

  • Inyoung Choi;Sathiyaraj Srinivasan
    • Journal of Species Research
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    • v.13 no.1
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    • pp.100-104
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    • 2024
  • The purpose of this study was to isolate and identify wild yeasts from soil samples collected in Daegu and Cheongju city, Republic of Korea. To identify the wild yeast strains, pairwise sequence comparisons of D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation test are done using API 20C AUX kit. All strains were assigned to the phylum Basidiomycota. Among 13 strains, 11 strains were previously reported, but two strains were unreported from the Republic of Korea. The two unrecorded yeast strains, GW1-3 and PG1-1-10C, belong to the genus Solicoccozyma (family Piskurozymaceae, order Filobasidiales, class Tremellomycetes). The two strains had oval-shaped and polar budding cells. This research showed the morphological and biochemical properties of the two unreported yeast species that had not officially reported in Korea.