WT1 gene promoter and enhancer in different cell lines in the transcriptional activity of.docVIP

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2017-05-05 发布于浙江
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WT1 gene promoter and enhancer in different cell lines in the transcriptional activity of.doc

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WT1 gene promoter and enhancer in different cell lines in the transcriptional activity of

WT1 gene promoter and enhancer in different cell lines in the transcriptional activity of Authors: Shao-Yan Hu, Chen Xing, Zhao Ye, Jian-Nong Cen, Gu Min, Fu Zheng-zheng, He Jun, Gu Weiying Abstract This study investigated the WT1 gene promoter and enhancer in different cell lines in the transcriptional activity of WT1 on gene regulation for the conduct of gene therapy of leukemia foundation. In order to make EGFP reporter gene construct containing the WT1 gene promoter and enhancer of the recombinant expression vector; the use of liposome and electroporation technology recombinant plasmid transfected cell lines 13, including high expression of WT1 gene leukemia cell lines (K562 , NB4, THP  1, SHI  1), WT1 gene expression in leukemia cell lines with low (U937 and Jurkat); non-hematopoietic cell lines of high WT1 gene expression in MCF  7, T47D, 293 cell lines, and lower expression of WT1 gene The ECV304, SMMC7721, HT  29, SHG44 cell line; the use of flow cytometry detection of transgenic cells stably expressing EGFP mean fluorescence intensity. The average fluorescence intensity on behalf of promoter and (or) to enhance the transcriptional activity of the child. The results showed that: The recombinant construct containing WT1 gene promoter expression vector pEWP, as well as containing the WT1 gene promoter and enhancer of the expression vector pEWPA. Transcriptional activity of the promoter in terms of non-leukemia cell lines, ECV304 cells in EGFP mean fluorescence intensity of the highest, is an empty vector (pEGFP  1) of 16.54 ± 2.45-fold higher than that of leukemia cell lines (p lt;0.05) ; MCF  7 and followed SHG44 cells were 9.46 ± 1.10 and 7.29 ± 0.73-fold, HT  29 cells expressed the lowest, only 0.99 ± 0.02-fold. In human leukemia cell lines, K562 cells, the average EGFP fluorescence intensity of the highest, is the empty vector pEGFP  1 of 2.93 ± 0.27-fold, significantly higher than Jurkat and SHI  1 cell lines (p lt;0.05). The latte