Transactivation of the human NME5 gene by Sp1 in pancreatic cancer cells
- 作者：Fu Li<sup>asup><sup> ; sup><sup>1sup> ; Zhenzhou Jiang<sup>asup><sup> ; sup><sup>1sup> ; Ke Wang<sup>bsup> ; Jingjing Guo<sup>asup> ; Gang Hu<sup>asup><sup> ; sup><sup>bsup> ; Lixin Sun<sup>asup> ; Tao Wang<sup>asup> ; Xuzhen Tang<sup>bsup><sup> ; sup><sup>csup> ; Ling He<sup>csup> ; Jincheng Yao<sup>dsup> ; Danyi Wen<sup>bsup> ; Xiaoran Qin<sup>bsup><sup> ; sup><sup>xiaoran_qin@yahoo.comsup> ; Luyong Zhang<sup>asup><sup> ; sup><sup>lyzhangchina@hotmail.comsup>
- 关键词：NME5 ; non-metastatic cells 5 ; Sp1 ; specificity protein 1 ; Sp3 ; specificity protein 3 ; AP2 ; activating protein 2 ; T-Ag ; T-antigen ; wt ; wildtype ; mut ; mutated ; RACE ; rapid amplification of cDNA ends ; EMSA ; Electrophoretic Mobility Shift Assay ; ChIP ; chromat
- 刊名：Gene
- 出版年：2012
- 期刊代码：73_03781119
- 类别：bio
- 出版时间：25 July, 2012
- 卷：503
- 期：2
- 页码：200-207
- 文件大小：887 K
摘要
Non-metastatic cells 5 (NME5), a recently found gene belonging to the NDPK-like molecules gene family, is highly expressed in testis and some types of human cancer. Current studies have revealed diverse potential functions of NME5 and we have reported that NME5 is associated with innate resistance to gemcitabine in human pancreatic cancer cells in previous study. However, the mechanism underlying the transcriptional regulation of NME5 has not been elucidated yet. In this study, we analyzed the 5鈥?flanking region of the human NME5 gene and revealed its transcription start site (TSS) at 鈭?#xA0;35 bp relative to its translation start codon ATG. Using 5鈥?unidirectional deletion analysis, we demonstrated that the proximal promoter of NME5 is located within 鈭?#xA0;1051 bp to + 35 bp. Two functional GC-boxes (鈭?#xA0;300 bp and 鈭?#xA0;323 bp) were identified within the promoter region. Mutation of either GC-box led to significant reduction in NME5 promoter activity, whereas overexpression of Sp1 activated NME5 promoter activity in MIA PaCa-2 and 293T cells. In silico analysis predicted that transcription factor Sp1 binds to both GC-boxes, which were confirmed by EMSA and ChIP. In addition, we found that compared with MIA PaCa-2, Sp1 was highly expressed in PAXC002, a well characterized human pancreatic cancer cell line with innate gemcitabine resistance where NME5 was reported to be highly expressed, indicating that Sp1 induces NEM5 expression in PAXC002 cells. In conclusion, our study characterized for the first time the human NME5 promoter which is controlled by Sp1 transcription factor in pancreatic cancer.