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

BMC Gastroenterology

Lin Guo²  Xiang Gao¹  Hongling Wang²  Jingjing Feng¹  Ran Xiao¹  Hui Zheng¹  Renquan Lu²  Ying Chen² 

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

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

Others  :  1137277
DOI  :  10.1186/s12876-015-0243-0

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

【 摘 要 】

Background

We 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.

Methods

Quantitative 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.

Results

Intensive 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.

Conclusion

The 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】

   
2015 Chen et al.; licensee BioMed Central.

【 图 表 】

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