Effects of rs35100176 polymorphism in IPO8 promoter on gene expression / 中国病理生理杂志
Chinese Journal of Pathophysiology
;
(12): 1640-1644, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-456850
ABSTRACT
AIM:
To investigate the effect of rs 35100176 CCT insertion/deletion polymorphism in the promoter region of importin 8 ( IPO8) gene on its mRNA expression .METHODS:
A 342-bp fragment of IPO8 gene promoter con-taining the rs35100176 polymorphism was amplified from 49 DNA samples and sequenced .The IPO8 promoter fragments containing CCT 3-nucleotide insertion or deletion were amplified using the corresponding homozygote DNA samples .The PCR products were sequenced and inserted into the luciferase reporter vector pGL 3-Basic.Recombinant vectors were trans-fected into the cells by Fugene 6.0 and the expression of the reporter gene was detected by a dual-luciferase reporter assay system.The mRNA expression level of IPO8 was detected by real-time PCR in 3-nucleotide insertion or deletion homozygote cells.RESULTS:
The sequencing results showed that there were 3 kinds of genotypes in the rs35100176 polymorphism, CCT/CCT, CCT/-and -/-, and the gene frequencies were 18.37%, 55.10% and 26.53%, respectively.The re-combinant expression vectors pGL 3-3N Insertion and pGL3-3N Deletion were successfully constructed .The luciferase assay showed that pGL3-3N Insertion produced significantly lower luciferase activity than that by pGL 3-3N Deletion.Real-time PCR showed that HEK293 cells with 3-nucleotide insertion homozygote expressed relative lower IPO 8 mRNA than Saos-2 cells with 3-nucleotide deletion homozygote .CONCLUSION:
The CCT 3-nucleotide insertion variant decreases the pro-moter activity of IPO8, thus affecting the gene expression .
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Pathophysiology
Year:
2014
Type:
Article
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