The NFKB1 polymorphism (rs4648068) is associated with the cell proliferation and motility in gastric cancer

BMC Gastroenterology

Research Article

Xiang Gao¹  Ran Xiao¹  Hui Zheng¹  Jingjing Feng¹  Lin Guo²  Renquan Lu²  Ying Chen²  Hongling Wang³ 

[1] Department of Clinical Laboratory, Shanghai Cancer Center, Fudan University, 200032, Shanghai, China;Department of Clinical Laboratory, Shanghai Cancer Center, Fudan University, 200032, Shanghai, China;Department of Oncology, Shanghai Medical College, Fudan University, 200032, Shanghai, China;Department of Oncology, Shanghai Medical College, Fudan University, 200032, Shanghai, China;

关键词: NFKB1;    Polymorphism;    Gastric cancer;    Susceptibility;    Single nucleotide polymorphism;   

DOI  :  10.1186/s12876-015-0243-0

 received in 2014-09-27, accepted in 2015-01-29,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundWe have demonstrated previously that NFKB1 single nucleotide polymorphism (SNP) rs4648068 GG homozygote was associated with the increased risk of gastric cancer in Chinese Han population. In this study, we constructed the recombinant plasmid pGL3-AA, pGL3-GG, pGL3-AA-NFKB and pGL3-GG-NFKB to investigate the function of rs4648068 by cell biology experiments.MethodsQuantitative real-time PCR was used to detect NFKB1 SNP rs4648068 genotype in the patients with gastric cancer. Anti-NF-κB1 p50 polyclonal antibodies were used for immunohistochemical analysis of the tissue specimens. The subsection of NFKB1 containing the promoter site and adjacent three consecutive exons were obtained by PCR technique and subcloned into the vector pGL3-Basic. Dual-Luciferase reporter assay was used to detect the transcriptional activity of the constructed promoter. Effects of transcription factor NFKB1 on C/EBPβ expression were determined by chromatin immunoprecipitation and Western analysis. Furthermore, proliferation and invasion ability of the transduced cell were also measured and compared.ResultsIntensive staining for p50 expression was observed in the tissues of GG genotype patients, compared with those of GA group and AA genotype patients. The transcriptional activity of rs4648068 (A > G) by dual-Luciferase reporter assay suggested that the luciferase activity of homozygote group (pGL3-GG) was greater than that of the control (pGL3-AA), especially at the stimulation of LPS. We found that the luciferase activity was also influenced by pGL3-GG levels. The effects of NFKB1 rs4648068 were enhanced by rs4648065 on the transduced cells. The interaction between NFKB1 promoter nucleotide sequence and C/EBPβ was regulated by the functional SNP rs4648068 in SGC-7901 cells. Our data indicated that the transduction of pGL3 expression plasmid pGL3-GG-NFKB improved the proliferation and motility of gastric cancer cells. Correspondingly, the homozygote GG of SNP rs4648068 strengthened the transcriptional activity of NFKB1 and influenced the cell biological activity.ConclusionThe transcriptional activity of NFKB1 was associated with SNP rs4648068, and this functional SNP site has the important effects on cell proliferation and motility.

【 授权许可】

Unknown   
© Chen et al.; licensee BioMed Central. 2015. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.