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Primary Study of the Expression Relationship between Humanβ1,3-N-acetylglucosaminyltransferase Gene and Transcription Factor Ets-1

Author: XuZhengRong
Tutor: WuShiLiang；ChouZuo
School: Suzhou University
Course: Biochemistry and Molecular Biology
Keywords: β3GnT Ets-1 Leukemia Chromatin immunoprecipitation
CLC: Q75
Type: Master's thesis
Year: 2010
Downloads: 13
Quote: 0
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Abstract


Purpose: beta1 ,3-N-acetyl glucosamine glycosyltransferase 8 (β3GnT8) in our laboratory by in silico cloning of a novel human glycosyltransferase homologous sequence comparison and belongs to the the beta3 sugar glycosyltransferase big family. , Scholars reported, β3GnT2 and β3GnT8, forming a heterodimer, the catalytic activity of this compound in vitro than the two respective enzyme activity was significantly enhanced, Further research showed that the transcription factor Ets-1 can regulate the expression of several glycosyltransferases This article aims by to study four leukemia cell lines β3GnT2, β3GnT4, β3GnT8 transcription factor Ets-1 in ATRA, TPA-induced differentiation of expression, and then explore their functional relevance in leukemia. Method: 1. Using RT-PCR and real-time quantitative PCR from mRNA level research β3GnT2, β3GnT4, β3GnT8 and transcription factor Ets-1 expression; chromatin immunoprecipitation (ChIP) was used to detect transcription factor Ets-1 to β3GnT2 β3GnT4 β3GnT8 gene expression and regulation; 3. immunofluorescence β3GnT2 the β3GnT8 with transcription factor Ets-1 in leukemia cell line K562 positioning expression. Results: Compared with control group, four cell lines after ATRA, TPA-induced differentiation β3GnT2 β3GnT4 β3GnT8 of Ets-1 expression were improved, β3GnT2 β3GnT8 changes in gene expression consistent and Ets 1 expression variation and β3GnT2 and β3GnT8 of gene expression variation; 2 from ChIP products for the purpose of gene amplification, the amplified ChIP products in K562 cells could be amplified β3GnT2 the β3GnT8 gene, but not amplified gene β3GnT4; immunofluorescence detection β3GnT2 and the perinuclear cytoplasm β3GnT8 have expression, transcription factor Ets-1 only in the nuclear and perinuclear expression, CY3 marked red fluorescence FITC-labeled green fluorescent overlap, the overlapping part of the yellow fluorescence. Conclusion: Compared with the control group, dosing β3GnT2 β3GnT8 gene expression trends and ETS-1 expression variation, and β3GnT2 β3GnT8 gene expression variation. Chromatin immunoprecipitation and immunofluorescence confirmed the transcription factor Ets-1 with glycosyltransferase β3GnT2, β3GnT8 combination. Therefore, the transcription factor Ets-1 may be involved in mediated regulation of gene expression β3GnT2 β3GnT8 raised β3GnT2 and β3GnT8 of gene expression of the transcription factor.

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