Expression quantitative trait loci in long non‐coding RNA ZNRD1‐AS1 influence both HBV infection and hepatocellular carcinoma development

Abstract

Zinc ribbon domain containing 1 (ZNRD1), cloned from human leukocyte antigen (HLA) region, may play integral roles in diverse processes including immune response against HBV infection and hepatocarcinogenesis. ZNRD1-AS1 (ZNRD1 antisense RNA 1) may be an important regulator of ZNRD1. By bioinformatics analyses, we identified that several single nucleotide polymorphisms (SNPs) in ZNRD1-AS1 may be expression quantitative trait loci (eQTLs) for ZNRD1. In this study, we hypothesized that these eQTLs SNPs in ZNRD1-AS1 may influence both chronic HBV infection and hepatocellular carcinoma (HCC) development. We designed a case-control study of 1300 HBV-positive HCC patients, 1344 HBV persistent carriers and, 1344 HBV natural clearance subjects to test the associations of three ZNRD1 eQTLs SNPs (rs3757328, rs6940552 and, rs9261204) in ZNRD1-AS1 with the risk of both chronic HBV infection and HCC. Logistic regression analyses in additive genetic model showed that variant alleles of all the three SNPs increased host HCC risk, whereas variant allele of rs3757328 was associated with HBV clearance. Moreover, the haplotype containing variant alleles of the three SNPs was significantly associated with both HCC development (adjusted OR = 1.18, 95% CI = 1.01-1.38, P = 0.035) and HBV clearance (adjusted OR = 0.83, 95% CI = 0.71-0.96, P = 0.013), when compared with the most frequent haplotype. In vitro experiments showed that ZNRD1 knockdown inhibited the expression of HBV mRNA and promoted proliferation of HepG2.2.15 cells. These findings suggest that ZNRD1 regulatory SNPs may be susceptibility makers for risk of both chronic HBV infection and HCC.

Strong evidence for LncRNA ZNRD1-AS1, and its functional Cis- eQTL locus contributing more to the susceptibility of lung cancer

“…Wen et al [7] first reported that rs9261204 and rs6940552 in lncRNA ZNRD1-AS1 may influence not only chronic HBV infection, but also hepatocellular carcinoma (HCC) development. The results of genetic associated study showed that rs3757328 were statistically associated with the lower gene expression of lncRNA ZNRD1 and high gene expression of ZNRD1-AS1 [7]. In current study, SNP rs9261204 was significantly associated with lung cancer and bladder cancer risk.…”

“…Using database RegulomeDB [19], which guides interpretation of regulatory variants in the human genome, we selected rs3757328, rs6940552, and rs9261204 as the tagSNPs of the Cis- eQTL for ZNRD1 in ZNRD1-AS1 gene region as previously [6, 7]. The genotyping were determined by using a polymerase chain reaction–restriction fragment length polymorphism method (PCR-RFLP).…”

“…Long non-coding RNAs (LncRNAs) has been confirmed to be associated with susceptibility, invasion, metastasis, and overall and disease-free survival of many cancers [9, 10]. Recently, two studies also evaluated the association of expression quantitative trait loci (eQTL) in LncRNA ZNRD1-AS1, a LncRNA in the upper region of ZNRD1 gene which could down-regulate the gene expression of ZNRD1, with the susceptibility and development of cervical cancer and hepatocellular carcinoma [6, 7]. However, no association study has conducted to evaluate the relationship between gene expression of ZNRD1 and lung cancer up to present, either in vivo or in vitro studies.…”

Long noncoding RNAs in hepatocellular carcinoma: Novel insights into their mechanism

“…By viewing dozens of research articles, we found that HULC can accelerate the growth of HCC via downregulating its neighbor gene p18 (known as eukaryotic translation elongation factor 1, EEF1E1 or AIMP3) and can disturb the circadian rhythm of HCC via upregulating oscillator CLOCK [47] . 1300 HBV-positive HCC patients was done [93] . The study found out that ZNRD1 antisense RNA 1 (ZNRD1-AS1) is a crucial regulator of ZNRD1 (human zinc ribbon domain containing 1).…”

“…ZNRD1 is involved in DNA damage and repair via regulating the expression of excision repair cross-complementing 1 (ERCC1) [96] , to restrain cell proliferation and to regulate the expression of miRNAs in cancers [97,98] . Furthermore, ZNRD1 eQTLs SNPs in lncRNA ZNRD1-AS1 have an increased risk for persistent HBV-carriers HCC but a protective influence against chronic HBV infection [93] . As a result, the different roles which ZNRD1-AS1 play make a difference in the treatment of HBV-positive HCC patients and HBVnegative HCC patients.…”