Single nucleotide polymorphism in the microRNA-199a binding site of HIF1A gene is associated with pancreatic ductal adenocarcinoma risk and worse clinical outcomes - PubMed (original) (raw)

Single nucleotide polymorphism in the microRNA-199a binding site of HIF1A gene is associated with pancreatic ductal adenocarcinoma risk and worse clinical outcomes

Xiuchao Wang et al. Oncotarget. 2016.

Abstract

Hypoxia-inducible factor-1 alpha (HIF-1α) is over-expressed in many cancers including pancreatic ductal adenocarcinoma (PDAC) and correlated with poor prognosis. We aim to determine the effect of germline genetic variants on the regulation of the homeostasis of the miRNA-gene regulatory loop in HIF1A gene and PDAC risk. HIF1A rs2057482 single nucleotide polymorphism (SNP) was genotyped in 410 PDAC cases and 490 healthy controls. The CC genotype SNP HIF1A is significantly correlated with PDAC risk (OR = 1.719, 95% CI: 1.293-2.286) and shorter overall survival (OS, P<0.0001) compared with the CT/TT alleles group. The C/T variants of rs2057482, a SNP located near the miR-199a binding site in HIF1A, could lead to differential regulation of HIF1A by miR-199a. Specifically, the C allele of rs2057482 weakened miR-199a-induced repression of HIF-1α expression on both mRNA and protein levels. In the PDAC tissue, individuals with the rs2057482-CC genotype expressed significantly higher levels of HIF-1α protein than those with the rs2057482-CT/TT genotype (P<0.0001). Both the CC genotype of SNP HIF1A and increased HIF-1α expression are significantly associated with shorter OS of patients with PDAC. After adjusted by TNM staging, differentiation grade, and the levels of CA19-9, both SNP HIF1A and HIF-1α expression retained highly significance on OS (P<0.0001). Taken together, our study demonstrates that host genetic variants could disturb the regulation of the miR-199a/HIF1A regulatory loop and alter PDAC risk and poor prognosis. In conclusion, the rs2057482-CC genotype increases the susceptibility to PDAC and associated with cancer progression.

Keywords: hypoxia-inducible factor-1 alpha (HIF-1α); microRNA-199a (miR-199a); pancreatic ductal adenocarcinoma (PDAC); single nucleotide polymorphism (SNP).

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Conflict of interest statement

CONFLICTS OF INTEREST

No potential conflicts of interest were disclosed.

Figures

Figure 1

Figure 1. The association between SNP of HIF1A and survival rate in patients with PDAC

A. Kaplan–Meier survival curves of overall survival (OS) of 269 PDAC patients with the CC genotype verses the CT/TT phenotypes. B. Kaplan–Meier curves of OS for 141 advanced PDAC patients with the CC genotype verses the CT/TT phenotypes. C, D. OS (C) and relapse-free survival (RFS) (D) of 128 PDAC patients who undergoing surgical resection, presented as CC verses CT/TT genotype groups. Data were analyzed by the log-rank test and the Kaplan–Meier curves were generated by using GraphPad Prism software.

Figure 2

Figure 2. Relative expression levels of miR-199a and HIF-1α in PDAC tissues

A. Immunohistochemical staining of HIF-1α in PDAC with low-, medium- and high-levels of HIF-1α expression; B. The frequency distribution of levels of HIF-1α expression between CC and CT/TT genotype groups (n = 60, *P < 0.05); C. Levels of miR-199a expression in PDAC tissues of CC or CT/TT genotype groups (n=60); D. Comparison of HIF-1α protein expression in samples with the CC genotype and CT/TT genotypes (n = 60, **P< 0.01).

Figure 3

Figure 3. miR-199a differential regulation of the SNP rs2057482

A. The predicted secondary structure of the HIF-1α mRNA. The secondary structures of the 3′UTR of HIF1A were predicted by inputting two 302-nt long DNA sequences centering rs2057482 into RNA fold, with either the rs2057482-C (left) or rs2057482-T (right) allele. The figures and the values of minimum free energy (MFE) were generated by RNAfold (

http://rna.tbi.univie.ac.at

); B. Schematic representation of the sequences of human miR-199a and its target site the 3′ UTR of HIF1A. The rs2057482 SNP is located 8 nucleotides (nt) downstream of the “seed complementary sequence” of miR-199a in HIF1A. C, D. Effects of rs2057482 genotypes on the expression of HIF1A gene in HEK-293T and Panc-1 cell lines. Forty-eight hours after transfection of the reporter gene and miR-199a mimic, the roles of construct with C or T allele on relative luciferase activity were compared in HEK-293T and Panc-1 cell lines (*P <0.05, **P <0.01). Data shown are mean ± SD of three independent experiments.

Figure 4

Figure 4. The role of miR-199a in the regulation of HIF1A transcription

The variant rs2057482 is a change of C-to-T located at 8bp downstream of the binding site of miR-199a in the 3 UTR of the HIF1A gene. MiR-199a is processed in pancreatic ductal epithelial cells and binds to the 3′UTR of the HIF1A gene. The rs2057482 C allele affects the miR-199a binding site at HIF1A 3′UTR, which damages the binding ability of miR-199a and results in the reduced degradation of HIF-1αmRNA and subsequently increases its translation, which may enhances PDAC risk and the development of PDAC.

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References

    1. Hidalgo M. Pancreatic cancer. N Engl J Med. 2010;362:1605–1617. - PubMed
    1. Song F, He M, Li H, Qian B, Wei Q, Zhang W, Chen K, Hao X. A cancer incidence survey in Tianjin: the third largest city in China-between 1981 and 2000. Cancer Causes Control. 2008;19:443–450. - PubMed
    1. Daemen A, Peterson D, Sahu N, McCord R, Du X, Liu B, Kowanetz K, Hong R, Moffat J, Gao M, Boudreau A, Mroue R, Corson L, et al. Metabolite profiling stratifies pancreatic ductal adenocarcinomas into subtypes with distinct sensitivities to metabolic inhibitors. Proc Natl Acad Sci U S A. 2015;112:E4410–4417. - PMC - PubMed
    1. Hollingsworth MA, Yeh JJ, Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur BK, Becker A, Hoshino A, Mark MT, Molina H, Xiang J, et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat Genet. 2015;17:816–826. - PMC - PubMed
    1. Wang X, Liu Y, Ren H, Yuan Z, Li S, Sheng J, Zhao T, Chen Y, Liu F, Wang F, Huang H, Hao J. Polymorphisms in the hypoxia-inducible factor-1alpha gene confer susceptibility to pancreatic cancer. Cancer Biol Ther. 2011;12:383–387. - PubMed

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