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Journal of Medical Virology 85:808814 (2013)

Association of Polymorphism in MicroRNA 219-1


With Clearance of Hepatitis B Virus Infection
Jae Youn Cheong,1 Hyoung Doo Shin,2 Yoon Jun Kim,3* and Sung Won Cho1
1

Department of Gastroenterology, Ajou University School of Medicine, Suwon, Republic of Korea


Department of Life Science, Sogang University, Seoul, Republic of Korea
3
Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine,
Seoul, Republic of Korea
2

Polymorphisms in the primary microRNA region may be associated with natural course of
hepatitis B virus (HBV) infection. This study
evaluated if the mircoRNA 219-1 (miR-219-1)
polymorphism can inuence the susceptibility
towards persistence of HBV infection and the
progression to hepatocellular carcinoma
(HCC) in patients with chronic HBV infection.
A total of 1,439 individuals having either past
or present evidence of HBV infection were enrolled for the study. The subjects were divided into four groups; (1) spontaneous recovery
(n 404), (2) chronic HBV carrier (n 313),
(3) chronic HBV carrier with cirrhosis
(n 305), and (4) hepatocellular carcinoma
(n 417). Genotyping was performed at three
polymorphic variants (rs421446, rs107822, and
rs213210) in the pri-miRNA region of miR-219-1.
The rs421446 T allele was found to be strongly
associated with HBV clearance (OR 0.73,
P 0.0005 in a codominant model and
OR 0.67, P 0.0009 in a dominant model,
OR 0.69, P 0.04 in a recessive model, respectively). The rs107822 G allele was also
found to be associated with HBV clearance
(OR 0.79, P 0.008 in a codominant model
and OR 0.72, P 0.01 in a dominant model,
respectively). In haplotype analysis, ht2 (T-G-T)
and ht1 (C-A-C) were found to be in signicant
association with the clearance of HBV. However,
no signicant association was observed between miR-219-1 polymorphism and the risk of
HCC occurrence. This result suggests that polymorphisms in the pri-miRNA region of miR-2191 might be a genetic factor for HBV clearance
after infection. J. Med. Virol. 85:808814,
2013. 2013 Wiley Periodicals, Inc.
KEY WORDS: polymorphism;
microRNA219; hepatitis B virus; hepatocellular carcinoma
2013 WILEY PERIODICALS, INC.

INTRODUCTION
Hepatitis B virus (HBV) infection is a major global
health problem for more than 350 million HBV carriers around the world [Iino, 2002; Lok and McMahon,
2007; Suk et al., 2012]. The clinical outcome of the
infection varies widely and may include spontaneous
recovery, the presence of inactive hepatitis B surface
antigen (HBsAg) carriers, chronic hepatitis, liver cirrhosis, or hepatocellular carcinoma (HCC). Perinatal
transmission of HBV is the major mode of infection in
HBV-endemic areas, including Korea. The most signicant factor affecting the chronicity of HBV infection is the age at time of infection; the chronicity in
Korean patients who are vertically infected seems to
be determined by host factors or genetic differences
rather than viral factors such as variations in virulence of the viral strains [Stevens et al., 1975; Coursaget et al., 1987; Tassopoulos et al., 1987]. Genetic
factors are also likely to modify the risk of HCC development among patients with HBV infection in cases
where family history of HCC is a well-known risk factor [Yu et al., 2000]. Recent studies by the authors
reported that genetic variations such as single nucleotide polymorphism (SNP) of transforming growth factor beta receptor III gene and tumor necrosis factor
alpha gene are signicantly associated with the risk

Grant sponsor: SNUH Research Fund; Grant number:


03-2012-021; Grant sponsor: Basic Science Research Program;
Grant number: 2010-0009943; Grant sponsor: Basic Research
Laboratory Program, National Research Foundation of Korea;
Grant sponsor: Ministry of Education, Science and Technology;
Grant number: 2010-0001200.
*Correspondence to: Yoon Jun Kim, M.D, Department of
Internal Medicine, Seoul National University College of Medicine, 103 Daehak-ro, Jongno-gu, Seoul 110-799, Republic of
Korea. E-mail: yoonjun@snu.ac.kr
Accepted 14 January 2013
DOI 10.1002/jmv.23551
Published online in Wiley Online Library
(wileyonlinelibrary.com).

MicroRNA 219-1 and Hepatitis B Virus Infection

of HCC occurrence or the clearance of HBV [Kim


et al., 2003, 2011].
MicroRNAs (miRNAs) are small, single-stranded,
2123 nucleotide-long non-coding RNA that functions
to modulate protein expression [Lagos-Quintana
et al., 2001]. Recent estimates indicate that miRNAs
regulate at least 30% of all protein-coding genes,
building complex regulatory networks that control
almost every cellular process [Filipowicz et al., 2008].
Association studies using SNPs in miRNA genomic
regions might be of help to evaluate the involvement
of miRNAs in disease. Sequence variations in miRNA
genes including pri-miRNAs, pre-miRNAs, could
potentially inuence the processing and/or target
selection of miRNAs [Duan et al., 2007]. In addition,
SNPs located within miRNA target sites have been
shown to affect the expression of the target gene and
contribute to susceptibility to human disease [Borel
and Antonarakis, 2008]. Xu et al. [2008] have
reported that polymorphisms in miRNA and their
targets are useful in predicting the risk of HCC occurrence. A G > C polymorphism in miR-146a precursor
(rs2910164) has been reported to predict HCC
development.
The function of mircoRNA 219-1 (miR-219-1) has
not been studied extensively. Recent report showed
that miR-219 modulates N-methyl-D-aspartate receptor mediated neurobehavioral dysfunction [Kocerha
et al., 2009]. Another investigation indicates that
miR-219 plays an important role in oligodendrocyte
differentiation [Dugas et al., 2010]. However, the biological roles of miR-219 in disease progression in
patients with HBV infection are not elucidated.
This study evaluated if the miR-219-1 polymorphisms can inuence the susceptibility towards persistence of HBV infection and the progression to HCC
in patients with HBV infection.
MATERIALS AND METHODS
Study Patients
To serve as primary patients for the casecontrol
study, a total of 1,439 individuals having either past
or present evidence of HBV infection were recruited
for the present study. The selected subjects were the
ones, who had enrolled in the outpatient clinic of the
liver unit and the Center for Health Promotion Center
at Seoul National University Hospital and Ajou University Hospital. Diagnoses of chronic carriers and
spontaneously recovered individuals were established
by repeated seropositivity for the hepatitis B surface
antigen (HBsAg; Enzygnost1 HBsAg 5.0; Dade
Behring, Marburg, Germany) over a 6-month period,
and for both anti-HBs (antibody to hepatitis B surface
antigen; Enzygnost Anti-HBs II; Dade Behring) and
anti-HBc (antibody to hepatitis B core antigen; ABCorek; DiaSorin s.r.l., Saluggia, Italy) of the IgG type
without HBsAg, respectively. The chronic carrier
group was assessed further for disease progression to
cirrhosis or HCC. All of the patients in the chronic

809

carrier group had undergone regular medical followups and had been evaluated with serum alphafetoprotein level, abdominal ultrasonography, and/or
a four-phase spiral liver CT scan more than twice a
year to detect early stages of HCC. Based on the clinical decision, dynamic contrast enhanced abdominal
MRI, bone scans, and chest CT, brain MRI, brain CT,
hepatic angiography, or PET scan was also carried
out in some of the patients.
Diagnosis of HCC was based on imaging ndings of
nodules larger than 1 cm showing intense arterial uptake followed by washout of contrast in the venousdelayed phases in four-phase multi-detector CT scan
or dynamic contrast enhanced MRI and/or biopsy
[Bruix and Sherman, 2011]. Cirrhosis of the liver, on
the other hand, was diagnosed pathologically or the
diagnosis was based on the clinical evidence of portal
hypertension such as the presence of visible collateral
vessels on the abdominal wall, esophageal varies on
esophagogastroscopy, palpable splenomegaly, and sonographically denite ndings of cirrhotic liver or ascites.
Exclusion criteria for the study patients included
the following: (i) tested positive for anti-HBs but not
for anti-HBc; (ii) tested positive for anti-HCV or antiHIV (Genedia1; Greencross Life Science, Yongin-shi,
Korea, HCV13.2; Dong-A Pharmaceutical, Seoul,
Korea); (iii) average alcohol consumption of 10 g/day
or an average number of 1 cigarette pack/s smoked
daily assessed through individual interviews; and (iv)
incurrence of other types of liver diseases such as
autoimmune hepatitis, toxic hepatitis, hemochromatosis, Wilsons disease, non-alcoholic steatohepatitis,
primary biliary cirrhosis, or BuddChiari syndrome.
None of the patients had a previous history of immunosuppressant or anti-viral treatment. Finally, written consent was secured from the patients prior to
conducting the study and ethical approval was
obtained from the Institutional Review Board for Human Research at Seoul National University Hospital
and Ajou University Hospital.
Genotyping of miR-219-1 Genome Region
Polymorphisms
Using the Wizard genomic DNA purication kit
(Promega, Madison, WI), genomic DNA was extracted
from patients peripheral blood samples. The SNP
genotyping was performed using the TaqMan1[Livak,
1999] assay in the ABI prism 7900HT sequence detection system (Applied Biosystems, Foster City, CA).
Genotyping quality control was performed in 10% of
the samples by duplicate checking (rate of concordance in duplicates 100%). Assay IDs of rs421446,
rs107822, and rs213210 were C__27015692_10,
C___2215075_20 and C___2215074_10, respectively
(Applied Biosystems).
Statistical Analyses
To determine the association of rs421446, rs107822,
and rs213210 with HBV clearance and HCC
J. Med. Virol. DOI 10.1002/jmv

810

Cheong et al.

occurrence, the odds ratio (95% Condence Interval)


was calculated using logistic analysis adjusted for age
(continuous value) and sex (male 0, female 1) as
covariates to eliminate or reduce any confounds that
might inuence the ndings. Data was managed and
analyzed using the Statistical Analysis System (SAS)
version 9.1 (SAS, Cary, NC). Using the PHASE
algorithm, version 2.0 [Stephens et al., 2001], haplotypes were then inferred from the genotyped SNPs.
Lewontins D0 (jD0 j) and the linkage disequilibrium
(LD) coefcient r2 were then examined using the
Haploview algorithm [Barrett et al., 2005] to measure
the LD between all pairs of Biallelic loci.

respectively (Table II). No signicant deviation from


the HardyWeinberg equilibrium was observed in all
the polymorphisms (P > 0.05, Table II).
Linkage disequilibrium coefcient (jD0 j and r2)
among the three SNPs was calculated based on the
genotypes of whole study subjects in this study. Pairwise comparisons between these three polymorphisms
revealed complete LD (jD0 j 1 and r2 6 1; Fig. 1C).
Using the PHASE algorithm, four common haplotypes
of rs421446, rs107822 and rs213210 were inferred
from those SNPs. They were ht1 (C-A-C), ht2 (T-G-T),
ht3 (C-A-T), and ht4 (C-G-T) and used for haplotype
association analysis.

RESULTS
Clinical Profiles of the Study Patients
The clinical proles of the subjects enrolled for the
study are listed in Table I. A total of 1,439 Korean
subjects, having either past or present evidence of
HBV infection, were enrolled in this study and classied into two groups: spontaneously recovered
subjects (n 404) and chronic HBV infected patients
(n 1,035). Chronic patients infected with HBV
comprised of 313 chronic hepatitis B patients, 305
liver cirrhosis patients, and 417 HCC patients
(Table I). In chronic HBV infection, subjects with
more progressive diseases tended to be older in age
and had higher male to female ratio, lower positive
rate of hepatitis B e antigen (HBeAg), and higher positive rate of anti-HBe.
Genotype and Minor Allele Frequency of
pri-miRNA Region of miR-219-1 SNPs in Korean
Population
By direct sequencing, three SNPs were identied in
the pri-miRNA region of miR-219-1. Figure 1A depicts
schematic gene map of rs421446, rs107822, and
rs213210 in pri-miRNA region of miR-219-1 on chromosome 6p21.32. Black blocks represent miR-219-1,
HSD17B8, and RING1 genes. The frequency in parentheses was based on the genotyping data (n 1,439).
The minor allele frequencies of the three genotyped
polymorphisms were 0.345 (rs421446 C > T), 0.387
(rs107822 A > G) and 0.470 (rs213210 C > T),

Association Analysis of pri-miRNA Region of


miR-219-1 SNPs With Risk of Liver Disease in
Korean Population
The genotype frequencies of both the chronic HBV
carriers and the spontaneous recovered groups for
each polymorphism were analyzed using a logistic
regression model, controlling for age and sex as covariates. The rs421446 T allele was found to be strongly
associated with HBV clearance (OR 0.73, P
0.0005 in a codominant model and OR 0.67,
P 0.0009 in a dominant model, OR 0.69, P 0.04
in a recessive model, respectively). The rs107822 G allele was also found to be associated with HBV clearance (OR 0.79, P 0.008 in a codominant model
and OR 0.72, P 0.01 in a dominant model, respectively). The rs213210 T allele was found to be in marginal association with HBV clearance (Table III).
In haplotype analysis, ht2 (T-G-T) and ht1 (C-A-C)
were found to be signicantly associated with the
clearance of HBV. The ht2 (T-G-T) showed the strongest genetic effects in codominant model (OR 0.73,
P 0.0004) (Table III).
Subsequently, the relationship between haplotypes
or SNPs of miR-219-1 and the development of HCC
were investigated in chronic HBV carriers. In the
analysis of the HCC occurrence in patients with
chronic hepatitis or liver cirrhosis, no signicant association was observed between haplotypes or SNPs of
pri-miRNA region of miR-219-1 and HCC occurrence
(Table III).

TABLE I. Clinical Prole of the Study Patients


Chronic carrier
Clinical profile
No. of patients
Age (mean (range))
Sex (male/female)
HBeAg (positive rate) (%)
Anti-HBe (positive rate) (%)
HBsAg (positive rate) (%)
Anti-HBs (positive rate) (%)
J. Med. Virol. DOI 10.1002/jmv

Spontaneously
recovered

Hepatocellular
carcinoma

Liver cirrhosis

Chronichepatitis

404
53.1 (2881)
272/132
0
37.7
0
100

417
57.5 (2582)
279/48
25.7
65.8
100
0

305
50.8 (2290)
231/74
29.7
50.2
100
0

313
44.4 (1879)
238/75
37.1
44.9
100
0

MicroRNA 219-1 and Hepatitis B Virus Infection

811

Fig. 1. Physical map, haplotypes, and linkage disequilibrium of rs421446, rs107822, and rs213210.
A: Schematic gene map and SNPs in the pri-miRNA region of miR-219-1 on chromosome 6p21.32.
Black blocks represent miR-219-1, HSD17B8, and RING1 genes, respectively. The frequency in parentheses was based on the genotyping data (n 1,439). B: Haplotypes of rs421446, rs107822 and
rs213210. C: Linkage disequilibrium coefcient (jD0 j and r2) among the three SNPs.

DISCUSSION
This study shows that the genetic polymorphisms
and the haplotypes of miR 219-1 are associated with
clearance of HBV. To the best of our knowledge, this
study is the rst report on the associations between
miR 219-1 polymorphisms and the outcome of HBV
infection.
It is widely accepted that the most important factor
affecting the chronicity of HBV infection is age at
time of infection [Shimbo et al., 1997]. However, this
is not the sole factor determining the persistence of
HBV and the host genetic factors involving genetic
polymorphisms are believed to be responsible for

clinical outcomes of HBV infection, especially in the


population whose transmission route is vertical infection, which signies that almost all individuals had
been exposed to the virus at the same age. Recent
studies have revealed that HLA class I and II [Ahn
et al., 2000; Thio et al., 2003], TNF-a[Kim et al., 2003;
Cheong et al., 2006], interleukin 10 [Shin et al., 2003]
and other various kinds of cytokines contribute to the
variable clinical outcome of HBV infection [Kim et al.,
2010; Park et al., 2010]. The present study also
strengthens the possibility that genetic factor is involved in natural course of HBV infection.
The miRNAs are small non-coding transcripts that
can control expression of protein-coding mRNA at the

TABLE II. Genotype and Minor Allele Frequency of rs421446, rs107822, and rs213210 in Korean Population (n 1,439)
Chromosome

Coordinatea

Genotype (#samples)

rs421446

33282761

rs107822

33283553

rs213210

33283802

CC (602)
CT (676)
TT (157)
AA (519)
AG (720)
GG (196)
CC (396)
CT (732)
TT (310)

rs#

Minor allele
frequency

Heterozygosity

HWEb

0.345

0.452

0.138

0.387

0.475

0.078

0.470

0.498

0.319

Human genome build 36.


P-values of deviation from HardyWeinberg Equilibrium among spontaneously recovered group.

J. Med. Virol. DOI 10.1002/jmv

J. Med. Virol. DOI 10.1002/jmv

rs421446
rs107822
rs213210
ht1 (C-A-C)
ht2 (T-G-T)
ht3 (C-A-T)
rs421446
rs107822
rs213210
ht1 (C-A-C)
ht2 (T-G-T)
ht3 (C-A-T)
rs421446
rs107822
rs213210
ht1 (C-A-C)
ht2 (T-G-T)
ht3 (C-A-T)
rs421446
rs107822
rs213210
ht1 (C-A-C)
ht2 (T-G-T)
ht3 (C-A-T)
rs421446
rs107822
rs213210
ht1 (C-A-C)
ht2 (T-G-T)
ht3 (C-A-T)

rs#
0.325
0.372
0.456
0.456
0.324
0.083
0.325
0.379
0.454
0.454
0.325
0.074
0.325
0.379
0.454
0.454
0.325
0.074
0.320
0.362
0.461
0.459
0.318
0.095
0.353
0.399
0.497
0.497
0.349
0.095

Case
0.396
0.427
0.505
0.505
0.396
0.078
0.325
0.367
0.458
0.457
0.324
0.089
0.320
0.362
0.461
0.459
0.318
0.095
0.331
0.373
0.455
0.455
0.329
0.083
0.335
0.385
0.453
0.452
0.335
0.065

Control
0.73 (0.620.87)
0.79 (0.670.94)
0.83 (0.700.98)
0.83 (0.700.98)
0.73 (0.610.87)
1.07 (0.791.45)
1.06 (0.861.30)
1.11 (0.901.36)
1.01 (0.831.23)
1.02 (0.841.24)
1.06 (0.861.31)
0.76 (0.531.09)
1.09 (0.861.39)
1.14 (0.901.44)
1.01 (0.811.27)
1.02 (0.821.28)
1.10 (0.871.39)
0.72 (0.491.08)
0.97 (0.741.25)
0.98 (0.761.26)
1.02 (0.801.29)
1.01 (0.801.29)
0.96 (0.741.25)
1.05 (0.701.58)
1.06 (0.811.38)
1.05 (0.811.36)
1.18 (0.921.51)
1.18 (0.921.52)
1.04 (0.801.36)
1.49 (0.962.34)

OR (95%CI)

Codominant

0.0005
0.008
0.03
0.03
0.0004
0.64
0.62
0.33
0.91
0.87
0.59
0.13
0.46
0.27
0.90
0.85
0.43
0.11
0.79
0.89
0.89
0.92
0.77
0.81
0.69
0.72
0.20
0.19
0.77
0.08

P
0.67 (0.520.85)
0.72 (0.560.93)
0.74 (0.570.97)
0.82 (0.621.08)
0.66 (0.520.84)
1.12 (0.811.55)
0.95 (0.721.26)
1.02 (0.771.36)
0.94 (0.691.27)
1.13 (0.801.58)
0.96 (0.731.27)
0.76 (0.521.11)
1.02 (0.741.39)
1.06 (0.771.46)
0.93 (0.661.32)
1.14 (0.781.68)
1.03 (0.751.40)
0.73 (0.481.11)
0.90 (0.651.26)
0.98 (0.701.38)
1.04 (0.721.50)
1.00 (0.661.52)
0.89 (0.641.25)
1.04 (0.671.63)
1.09 (0.761.56)
1.01 (0.701.46)
1.15 (0.771.70)
1.36 (0.902.06)
1.07 (0.751.52)
1.56 (0.962.51)

OR (95%CI)

Dominant

0.0009
0.01
0.03
0.16
0.0007
0.49
0.74
0.88
0.68
0.49
0.77
0.16
0.92
0.74
0.69
0.50
0.87
0.15
0.55
0.90
0.85
0.99
0.51
0.85
0.63
0.95
0.50
0.14
0.73
0.07

0.69 (0.480.98)
0.77 (0.561.07)
0.82 (0.621.08)
0.74 (0.570.97)
0.68 (0.480.97)
0.59 (0.172.03)
1.47 (0.932.32)
1.43 (0.952.15)
1.12 (0.801.57)
0.95 (0.701.28)
1.48 (0.942.33)
0.40 (0.053.53)
1.47 (0.872.49)
1.52 (0.942.45)
1.14 (0.771.67)
0.95 (0.671.33)
1.47 (0.872.49)
0.28 (0.032.74)
1.14 (0.642.03)
0.97 (0.581.63)
1.01 (0.671.52)
1.03 (0.711.48)
1.15 (0.642.05)
1.28 (0.246.89)
1.03 (0.591.81)
1.17 (0.711.92)
1.36 (0.902.06)
1.15 (0.781.71)
1.02 (0.581.78)
1.49 (0.2010.99)

OR (95%CI)

Recessive

0.04
0.12
0.17
0.03
0.04
0.40
0.10
0.09
0.50
0.72
0.09
0.41
0.15
0.09
0.51
0.75
0.15
0.27
0.67
0.92
0.98
0.88
0.64
0.77
0.92
0.54
0.15
0.48
0.95
0.70

Bold values represent statistical signicance (P < 0.05).


Minor allele frequencies and P-values for logistic analyses of three alternative models (co-dominant, dominant, and recessive models) are shown.
HBV, hepatitis B virus; HCC, hepatocellular carcinoma; CH, chronic hepatitis; LC, liver cirrhosis; SR, spontaneously recovered; HBsAg, hepatitis B surface antigen; and HBeAg, hepatitis B
e antigen.

HBeAg clearance chronic (n 189)


versus cleared (n 428)

LC occurrence on CH LC (n 305)
versus CH (n 313)

HCC occurrence on LC HCC


(n 417) versus LC (n 305)

HCC occurrence HCC (n 417)


versus CH & LC (n 618)

HBV infection HBV (n 1,035)


versus SR (n 404)

Test group

Minor allele frequency

TABLE III. Association Analysis of rs421446, rs107822 and rs213210 on the pri-miRNA Region of miR-219-1 With Risk of Liver Disease in Korean Population
(n 1,439)

812
Cheong et al.

MicroRNA 219-1 and Hepatitis B Virus Infection

post-transcriptional level. Single miRNA molecules


are predicted to be capable of inuencing the expression of hundreds of different targeted genes [Bartel,
2009]. In recent years, much effort has been directed
towards understanding the role of SNPs present in
pri-, pre-, and mature miRNA and their inuences on
susceptibility to various diseases. So far, little is
known about the involvement of miRNAs during HBV
infection. Recently, a differential miRNAs expression
pattern was found in the livers of HBV and HCV
infected individuals with hepatocellular carcinoma
[Ura et al., 2009]. Another study has shown that a
liver specic micro RNA, miR-122, binds to a highly
conserved HBV pre-genomic RNA sequence and inhibits HBV gene expression and replication [Chen et al.,
2011]. This report suggested that miR-122 might
down-regulate HBV replication by binding to the viral
target sequence, thus contributing to the persistent
infection of HBV. In the current study, we identied
the association between SNPs of miR-219-1 and HBV
clearance. However, the biological role of miR-219-1
SNP is not assessed in current study, thus functional
signicance of miR-219-1 SNP in the natural course of
HBV infection is unclear. It remains to be elucidated
whether miR-219-1 function to modulate host immunity to eradicate virus during the early stage of infection or function through other mechanisms.
The miRNAs have emerged as important genomic
regulators with a key role in the development of cancer
[Boyd, 2008]. The potential role of miR-219 as an inhibitor of cellular proliferation has been indicated in previous research [Wong et al., 2008]. Recent investigation
showed miR-219-5p induced inhibition of HCC cell proliferation by targeting glypican-3 [Huang et al., 2012].
Other studies identied miR-219 as being downregulated in various cancers [Wong et al., 2008; Ferretti
et al., 2009; Izzotti et al., 2009]. However, the association studies between genetic variation in miR 219-1
and various tumors including oral premalignant lesion,
renal cell carcinoma and metastatic colon cancer
showed negative results. In current study, no signicant association was observed between genetic variations in pri-miRNA region of miR-219-1 and the
occurrence of HCC in patients with chronic HBV infection. However, there still exists the notation that the
other miRNA might inuence HCC development and
further studies are warranted on this subject.
In conclusion, it is stated that miR-219-1 SNP is
associated with clearance of HBV infection. This
result suggests that miR-219-1 polymorphism might
be an important genetic factor, which affects variable
outcome of HBV infection.
ACKNOWLEDGMENTS
The biospecimens from Ajou University Hospital
were provided by the Ajou Human Bio-Resource Bank
(AHBB), a member of the National Biobank of Korea,
which is supported by the Ministry of Health and
Welfare.

813

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