Primary Study Of The Expression Relationship Between Humanβ1,3-N-acetylglucosaminyltransferase Gene And Transcription Factor Ets-1

Object:β1,3-N-acetylglucosaminyltransferase 8 is a new member of glycosyltransferases we gained by electric cloning. We considered it belongs to beta-3 glycosyltransferase family .It is reported thatβ3GnT2 andβ3GnT8 can form a complex with enhanced enzymatic activity. It is also reported that there have been some relationship between glycosyltransferases and the transcription factor Ets-1. This thesis aims to base on studying the expression discrepancy ofβ3GnT2,β3GnT4,β3GnT8 gene and transcription factor Ets-1 in several different leukemia cells that be induced by ATRA,TPA, thereby study on the function relationship of them in leukemia cells.Methods: 1. RT-PCR and Real time-PCR was used to detect the mRNA expression discrepancy ofβ3GnT2,β3GnT4,β3GnT8 and transcription factor Ets-1;2.Chromatin Immunoprecipitation was used to detect the relationship ofβ3GnT2,β3GnT4,β3GnT8 gene and transcription factor Ets-1;3. Immunoprecipitation was used to detect the orientational expression of transcription factor Ets-1,β3GnT2 andβ3GnT8 in leukemia cells.Results: 1. The expression ofβ3GnT2,β3GnT4,β3GnT8 gene increased in several different leukemia cells that be induced by ATRA,TPA compared to control group. The expression discrepancy tendency ofβ3GnT2 was similar withβ3GnT8; 2.β3GnT2 andβ3GnT8 gene can be amplified in the product of Chromatin Immunoprecipitation, andβ3GnT4 gene can not be amplified; 3. As immunofluorescence assay indicated,β3Gnt2 andβ3GnT8 are expressed in the cytoplasma and around the nuclear, while transcription factor Ets-1 was only detected in the nuclear. The overlapped part of red CY3 and green FITC yielded yellow fluorescence.Conclusion: Compared to control group, the expression variation tendency ofβ3GnT2 was similar withβ3GnT8 and also similar with Ets-1, we suggested transcription factor Ets-1 may have influence on the expression ofβ3GnT2 andβ3GnT8, and may upregulated the expression ofβ3GnT2 andβ3GnT8.Transcription factor Ets-1 may have influence on the expression ofβ3GnT2 andβ3GnT8 after the Chromatin Immunoprecipitation and Immunofluorescence detected.