Comparison of SNP Polymorphism of Alcohol Metabolizing Related Enzyme Genes in Intoxicated Individuals and Nonalcoholic Population

Research Article

Austin J Forensic Sci Criminol. 2015; 2(3): 1031.

Comparison of SNP Polymorphism of Alcohol Metabolizing Related Enzyme Genes in Intoxicated Individuals and Nonalcoholic Population

Li Li*, Ping Xiang, Yan Liu, Yan Shi, Yuan Lin and Zhenmin Zhao

Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Science, China

*Corresponding author: Li Li, Shanghai Key Laboratory of Forensic Medicine, Institute of Forensic Science, Ministry of Justice, 200063, China

Received: July 16, 2015; Accepted: August 08, 2015; Published: August 11, 2015

Abstract

Objective: To compare the distributions of SNP allele and genotype about alcohol dehydrogenase 2 (ADH2), aldehyde dehydrogenase 2 (ALDH2) and CYP2E1 in intoxicated individuals and nonalcoholic population.

Methods: 100 individuals who were penalized duo to intoxicated driving were genotyped for 40 SNPs of ADH2,ALDH2 and CYP2E1 using multiplex PCR and Iplex chemistry on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer and compared with a reference group of 99 blood donors. Allele frequencies and genotype frequencies of 40 SNP loci were calculated and compared in the two groups.

Results: Among the 40 SNP loci, seven SNPs (e.g. rs698, rs2241894, rs1789915, rs13306164, rs671, rs28371746 and rs2515641) were polymorphic in the intoxicated population, but only six SNPs (e.g. rs698, rs2241894, rs13306164, rs671, rs28371746 and rs2515641) were polymorphic in the control individuals. Two SNP loci (rs671 in and rs2515641) were found to have a significant difference in frequency distribution between the two populations (p<0.01).

Conclusion: Among the above 40 SNP loci of ADH2,ALDH2 and CYP2E1 genes, rs671 and rs2515641 in the intoxicated population were found to differ in allele and genotype frequency distribution from the nonalcoholic population and may be related to alcohol intoxication.

Keywords: ADH; ALDH; CYP2E1; Single nucleotide polymorphism; Genetic polymorphism

Introduction

The metabolism of drug shows inter-individual variation and inter-ethnic variation. Because of the difference of Drug-Metabolizing Enzyme (DMEs) activity, the individuals can be divided into Poor Metabolizers (PMs), Extensive Metabolizers (EMs) and Ultra-Rapid Metabolizers (UMs) [1,2]. Therefore, it is usually a hard work to interpret drug related poisoning or death in forensic science [3]. Now punitive measures against people who drink and drive have been strictly enforced. In china, current standard for judging drunk drinking and drunken drinking is mainly based on the concentration of ethanol in blood. The driver whose alcohol concentration above 0.2mg/mL in blood is convicted of drunk drinking and the driver whose alcohol concentration above 0.8mg/mL in blood is convicted of drunken drinking, but, except for alcohol dehydrogenase (ADH), aldehyde dehydrogenase 2 (ALDH2) and cytochrome P450 2E1 enzyme (CYP2E1) has also been found to be involved in alcohol metabolism [4-7], which means that ALDH2 and CYP2E1 with gene polymorphism may also play dominant role in alcohol metabolism. Individuals with distinct genotypes are different in alcohol tolerance and show unequal behavioral responses after drinking [8]. Because of high concentration of acetaldehyde in blood transformed from ethanol, poor metabolism of acetaldehyde may lead to traffic accident even if the individual’s alcohol concentration below 0.2mg/mL in blood. The individual differences are mainly caused by the gene polymorphisms of ADH, ALDH and CYP2E1. It was confirmed that the presence of the less-active form of alcohol dehydrogenase- 1B encoded by ADH1B*1/*1 and active form of ALDH2 encoded by ALDH2*1/*1 increases the risk of alcoholism in East Asians [9]. SNPs were found to be related to the polymorphism of enzyme activity [8], base deletions, insertions, substitutions or transversions in genes of ADH, ALDH2 and CYP2E1 may lead to changes of amino acid sequences and result in enzyme activities lost or decreased or increased. In order to investigate SNP polymorphism of alcohol metabolizing- related enzyme genes in Chinese Han population, we genetyped 40 SNPs of ADH2, ALDH2 and CYP2E1 based on multiplex amplification and matrix-assisted laser desorption/ionization timeof- flight mass spectrometry (MALDI-TOF MS) [10,11] and compared the SNP polymorphism in intoxicated population and nonalcoholic population.

Materials and Methods

Selection of SNP loci and design of primers

Corresponding to sequences of genes coding for ADH2, ALDH2 and CYP2E1, 40 SNP loci (Table 1) were selected via NCBI website: https://www.ncbi.nlm.nih.gov/. Forty groups of primes were designed via Mass ARRAY Assay Design software (Sequenom, Inc.). Each group has three primes, including a pair of PCR primes and a single base extension prime (Table 2). Primers were synthesized by Shanghai Biological Engineering Technology Corporation.

Table 1: 40 SNPs in Alcohol Metabolizing Related Enzyme Genes.





  

    
Gene

    
SNP Loci

    
Variation

    
Gene

    
SNP Loci

    
Variation

  

  

    
ADH2

    
rs2066702

    
C/T

    
CYP2E1

    
rs35844228

    
C/G

  

  

    
 

    
rs55882921

    
C/T

    
 

    
rs72559710

    
A/C/G

  

  

    
 

    
rs41275697

    
C/T

    
 

    
rs28371740

    
A/G

  

  

    
 

    
rs1126440

    
C/T

    
 

    
rs60719153

    
C/T

  

  

    
 

    
rs41275699

    
A/G

    
 

    
rs56864127

    
A/G

  

  

    
 

    
rs67420531

    
A/G

    
 

    
rs60452492

    
A/G

  

  

    
ADH3

    
rs56247447

    
C/T

    
 

    
rs56040284

    
A/G

  

  

    
 

    
rs698

    
A/G

    
 

    
rs28371743

    
C/T

  

  

    
 

    
rs1042756

    
C/G

    
 

    
rs41299426

    
A/G

  

  

    
 

    
rs55717907

    
A/G

    
 

    
rs61710826

    
A/G

  

  

    
 

    
rs2241894

    
A/G

    
 

    
rs28371746

    
A/C

  

  

    
 

    
rs1789915

    
A/G

    
 

    
rs61644766

    
C/G

  

  

    
 

    
rs6490301

    
C/T

    
 

    
rs41299434

    
C/G

  

  

    
ALDH2

    
rs1064903

    
C/G

    
 

    
rs59981143

    
A/G

  

  

    
 

    
rs13306164

    
C/T

    
 

    
rs55897648

    
A/G

  

  

    
 

    
rs58280059

    
A/G

    
 

    
rs60207639

    
A/G

  

  

    
 

    
rs1062136

    
A/T

    
 

    
rs59656378

    
C/G

  

  

    
 

    
rs1064933

    
A/G

    
 

    
rs57702102

    
A/G

  

  

    
 

    
rs671

    
A/G

    
 

    
rs2515641

    
C/T

  

  

    
 

    
rs59868347

    
A/G

    
 

    
rs55982231

    
A/G

  










Table 1:  40 SNPs in Alcohol Metabolizing Related Enzyme Genes.

Table 2: 40 groups of primes used for SNP genotyping.





  

    
SNP_ID 

    
Forward primer (5’→3’) for PCR 

    
Reverse primer (5’→3’) for PCR 

    
Primer for extension reactions 

  

  

    
rs2066702

    
ACGTTGGATGGCATGTGGGTTGTCTAAATG

    
ACGTTGGATGCTCTATTGCCTCAAAACGTC

    
gCTTCTTTCCTATTGCAGTATC

  

  

    
rs55882921

    
ACGTTGGATGAAGAGTAAAGAAGGTATCCC

    
ACGTTGGATGCATGGGTTATTAACGCATCC

    
cccccAACTTGTGGCTGATTTTAT

  

  

    
rs41275697

    
ACGTTGGATGCAGTGGCCAAAATTGATGC

    
ACGTTGGATGGACCCATAACCAGTCGAGAA

    
AAAATTGATGCAGCCTC

  

  

    
rs1126440

    
ACGTTGGATGTGAACCTCCTGGTGCCATC

    
ACGTTGGATGAACCCGGAGAGCAACTACTG

    
TCCTGCAGGGTCCCCCG

  

  

    
rs41275699

    
ACGTTGGATGAACACTCTCCACGATGCCG

    
ACGTTGGATGATGACCACGTGGTTAGTGGC

    
CTGCCTCATGGCCTAAA

  

  

    
rs67420531

    
ACGTTGGATGTGTCTCTTCTTTCCTATTGC

    
ACGTTGGATGCTACAAGGGAAGGCATCTGT

    
TTCTTTCCTATTGCAGTATC

  

  

    
rs56247447

    
ACGTTGGATGATAAATGAAGGATTTGACC

    
ACGTTGGATGCGCTACTGTAGAATACAAAG

    
AGGATTTGACCTGCTTC

  

  

    
rs698

    
ACGTTGGATGAAGAAGTTTTCACTGGATGC

    
ACGTTGGATGAGAGCGAAGCAGGTCAAATC

    
CACTGGATGCATTAATAACAAAT

  

  

    
rs1042756

    
ACGTTGGATGCCAGTGAAAACTTCTTAGCC

    
ACGTTGGATGCTTCTTTTCAGGCTTTAAGAG

    
AAAGTCAGCCACAAGTTT

  

  

    
rs55717907

    
ACGTTGGATGCAGACCCATAACCAGTCGAA

    
ACGTTGGATGCCAAAATTGATGCAGCCTCG

    
CCATAACCAGTCGAAAATCCACA

  

  

    
rs2241894

    
ACGTTGGATGTCGGCGTCAGCACCTTCTC

    
ACGTTGGATGCGAGGCTGCATCAATTTTGG

    
CGGCGTCAGCACCTTCTCCCAGTACAC

  

  

    
rs1789915

    
ACGTTGGATGGTGATAAAGTCATCCCGCTC

    
ACGTTGGATGAAGCAGTAGTTGCTTTCTGG

    
TGTGGAAAATGCAGAATTTG

  

  

    
rs6490301

    
ACGTTGGATGTGACAAGAGGCGGCGGCCCA

    
ACGTTGGATGTCCGCTAGCCCGCTGCGAT

    
acggtGCCCGAAGCGGGCGGCAGC

  

  

    
rs1064903

    
ACGTTGGATGCTAGCCCGCTGCGATGTTG

    
ACGTTGGATGTGGCGGCGGCTGACAAGAG

    
TGCCGCCGCTTCGGGCCCCGCCTGG

  

  

    
rs13306164

    
ACGTTGGATGCATGGACGCATCACACAGG

    
ACGTTGGATGTAGGTCCGGTCCCGCTCGAT

    
ACGCATCACACAGGGGCCGGCTG

  

  

    
rs58280059

    
ACGTTGGATGCACGTTTCCAGTTGCCAAGG

    
ACGTTGGATGGCTGTTGTTTGTTGCAGTGG

    
CCATGCTTGCATCAGGAG

  

  

    
rs1062136

    
ACGTTGGATGAGGAGGACATCTATGATGAG

    
ACGTTGGATGTATCAAAGGGGTTCCCGAC

    
ggCATCTATGATGAGTTTGTGG

  

  

    
rs1064933

    
ACGTTGGATGTGTATGCCTGCAGCCCGTA

    
ACGTTGGATGACCCTTTGGTGGCTACAAGA

    
CCGTACTCGCCCAACTCCCGGCCACT

  

  

    
rs671

    
ACGTTGGATGCAGGTCCCACACTCACAGTT

    
ACGTTGGATGAGTTGGGCGAGTACGGGCTG

    
AGGTCCCACACTCACAGTTTTCACTT

  

  

    
rs59868347

    
ACGTTGGATGTTCCTCCTGCTGGTGTCCAT

    
ACGTTGGATGATGATGGGAAGCGGGAAAGG

    
gtTGTCCATGTGGAGGCAG

  

  

    
rs35844228

    
ACGTTGGATGCTTCCTTCACCGCCTTGTAG

    
ACGTTGGATGTTCGGGCCGGTGTTCACG

    
ATCACCACCATGCGCTGCGAGCCCA

  

  

    
rs72559710

    
ACGTTGGATGCTTCCTTCACCGCCTTGTAG

    
ACGTTGGATGGGCCGGTGTTCACGCTGTA

    
GTAGCCGTGCATCACCACCATG

  

  

    
rs28371740

    
ACGTTGGATGAAGGAAGCGCTGCTGGACTA

    
ACGTTGGATGTGTCCCTGTGCGCATGGAAC

    
GGACTACAAGGACGAGTTCTC

  

  

    
rs60719153

    
ACGTTGGATGATAATGGACCTACCTGGAAG

    
ACGTTGGATGTTTCCCCATCCCATAGTTCC

    
CCTACCTGGAAGGACATC

  

  

    
rs56864127

    
ACGTTGGATGTTTCCCCATCCCATAGTTCC

    
ACGTTGGATGATAATGGACCTACCTGGAAG

    
GTGGTCAGGGAAAACCGC

  

  

    
rs60452492

    
ACGTTGGATGGTTTTGTAGGCCAGCCTTTC

    
ACGTTGGATGGAAGAGGATGTCGGCTATGA

    
ACCCCACCTTCCTCATC

  

  

    
rs56040284

    
ACGTTGGATGGTTTTGTAGGCCAGCCTTTC

    
ACGTTGGATGGAAGAGGATGTCGGCTATGA

    
GACCCCACCTTCCTCATCGGCTGC

  

  

    
rs28371743

    
ACGTTGGATGTGATGAGAAGTTTCTAAGGC

    
ACGTTGGATGAGGGAGTGCTGAGTAGGTG

    
TAAGGCTGATGTATTTGTTTAA

  

  

    
rs41299426

    
ACGTTGGATGGTTGCATCCAGAAAAAAGTAG

    
ACGTTGGATGAAGTAGTGTAGAAAGCTGGG

    
ggTTCCCTCTCTAGCTTTAC

  

  

    
rs61710826

    
ACGTTGGATGAAAGAACAGGTCGGCCACAG

    
ACGTTGGATGCGGTATCACAGGAAAAGCAC

    
CACAGTCACGGTGATAC

  

  

    
rs28371746

    
ACGTTGGATGGAGAATCAGGAGCCCATATC

    
ACGTTGGATGTGTGGCCGACCTGTTCTTTG

    
AGCCCATATCTCAGAGTTGTGCTGGT

  

  

    
rs61644766

    
ACGTTGGATGAATTGACAGGGTGATTGGGC

    
ACGTTGGATGACCACAGCATCCATGTAGGG

    
AGCCGAATCCCTGCCAT

  

  

    
rs41299434

    
ACGTTGGATGTGCATGAGATTCAGCGGTTC

    
ACGTTGGATGCTCTGAAAATGGTGTCTCGG

    
cGTTCATCACCCTCGTGCCCT

  

  

    
rs59981143

    
ACGTTGGATGGGTATCCTCTGAAAATGGTG

    
ACGTTGGATGGAGATTCAGCGGTTCATCAC

    
GAAAATGGTGTCTCGGG

  

  

    
rs55897648

    
ACGTTGGATGTCAGGAAATTCTTGGTTGTC

    
ACGTTGGATGGTCTTTGTTTCTCCTAGGGC

    
tagcGTCCAGAGTTGGCACTA

  

  

    
rs60207639

    
ACGTTGGATGCTGGATCAGGAAATTCTTGG

    
ACGTTGGATGCTTTGTTTCTCCTAGGGCAC

    
GGTTGTCATACAAAACAGAG

  

  

    
rs59656378

    
ACGTTGGATGCTGGATCAGGAAATTCTTGG

    
ACGTTGGATGCTTTGTTTCTCCTAGGGCAC

    
agaacTTCTTGGTTGTCATACAAAA

  

  

    
rs57702102

    
ACGTTGGATGGTTTAAGCCAGAACACTTCC

    
ACGTTGGATGTCTCACCTGTGGAAAATGGC

    
cGAACACTTCCTGAATGAAA

  

  

    
rs2515641

    
ACGTTGGATGGCCAGAACACTTCCTGAATG

    
ACGTTGGATGTCTCACCTGTGGAAAATGGC

    
CCTGAATGAAAATGGAAAGTT

  

  

    
rs55982231

    
ACGTTGGATGTCATGAGCGGGGAATGACAC

    
ACGTTGGATGTATCGACCTCAGCCCTATAC

    
gcggtGAATGACACAGAGTTTGTAA

  










Table 2:  40 groups of primes used for SNP genotyping.

DNA extraction

Blood samples were collected from 100 unrelated Han individuals who were penalized duo to intoxicated driving. Another 99 blood samples for control were collected from nonalcoholic males. DNA was extracted according to the instructions of the blood genomic DNA Mini Kit (Sangon Biotech, Shanghai, China).

Multiplex PCR amplification

Aliquots of 1 μL DNA were amplified in a total volume of 5μL. Each reaction contained 0.625μL PCR buffer (10×) (Qiagen GmbH), 0.325μL 25 mmol/L MgCl2, 1μL dNTP (2.5mmol/L) (Tatara Inc.), 0.1μL of HotStarTaq polymerase (5U/μL), 0.95μL H2O, and 1μL the designed primers at their optimized concentrations. Using a Gene Amp PCR System 9700 (Applied Biosystems, Norwalk, CT), the reaction mixtures were incubated at 94°C for 15 min and then cycled 45 times through desaturation at 94deg;C for 20 s, annealing at 56deg;C for 30 s and extension at 72deg;C for 60 s and finally incubated at 72deg;C for 3 min. No-template controls were carried along in every plate to exclude contaminations.

SAP process

After PCR, the products were treated with hrimp alkaline phosphatase (SAP) to remove excess dNTPs. This dephosphorylation reaction contained 0.3μL SAP (1U/μL), 0.17μL SAP buffer (10×), 1.53μL H2O (Sequenom, Inc.) was carried out at 37deg;C for 40 min, and 85deg;C for 15 min.

Primer extension reactions

The PCR products were used as templates for the primer extension reactions. Extension reactions (final volume, 9μL) contained 0.2μL iPlex buffer (10×), 0.1μL iPlex termination mix, 0.0205μL iPlex enzyme, 0.7395μL H2O (all from Sequenom, Inc.), and 0.94μL extension primers at optimized concentrations. On a Gene Amp PCR System 9700 (Applied Biosystems, Norwalk, CT), extension reactions were performed at 94deg;C for 30 s followed by 40 cycles (94deg;C for 5 s, followed by 5 cycles of 52deg;C for 5 s, 80deg;C for 5 s); and finally 72deg;C for 3 min. The final nucleotide extension products were treated with a cationic exchange resin (AG® 50W-X8 Resin; Bio-Rad Inc.) for 30 min to remove salts. All reactions, including PCR amplification, shrimp alkaline phosphatase treatment, and base extension, were performed in 384 microtiter plates (Sequenom Inc.).

MALDI-TOF-MS

The reaction products were spotted onto the Mass ARRAY SpectroCHIP with an auto-spot arm (Sequenom, Inc.). The target plate was then inserted into the MALDI-TOF mass spectrometer of Mass ARRAY compact System (Sequenom, Inc.), and analysis was performed with 180 nitrogen laser shots for each sample. The mass range of the MS instrument was set at 3920–12023 Da. SNP loci was genotyped by Mass ARRAY Type Analyzer software version 4.0 (Sequenom, Inc.).

Statistical analysis

The data were analyzed with SPSS 13.0. The statistical information included genotype frequency, allele frequency and p values.

Results and Discussion

Evaluation of the MALDI-TOF MS genotyping assay

To evaluate the established SNP genotyping assay, we analyzed 100 genomic DNA samples from individuals of Chinese origin who had previously been genotyped for some of the SNPs by TaqMan assay and got consistent results.

Polymorphisms of SNPs

Among the 40 SNP loci in intoxicated population, three SNPs (rs698, rs2241894, rs1789915) in ADH3 gene, two SNPs (rs13306164, rs671) in ALDH2 gene, two SNPs (rs28371746, rs2515641) in CYP2E1 gene were found to be polymorphic, i.e. the minor allele frequency(MAF) of each SNP locus was above 1%, while others were not ( the MAF of them was below 1%). Among the 40 SNP loci in control population, the seven SNPs except rs1789915 were found to be polymorphic. The observed genotype and allele frequencies in intoxicated population and control population were shown in table 3 and 4 respectively.

Table 3: Genotype frequencies of 40 SNPs in intoxicated population (n=100) and control population (n=99).





  

    
Gene

    
SNP Loci

    
Variation

    
Genotype frequencies in intoxicated population

    
Genotype frequencies in control population

  

  

    
ADH2

    
rs2066702

    
C/T

    
CC:1.0000

      
CT: 0.0000

      
TT:0.0000

    
CC:1.0000

      
CT: 0.0000  TT:0.0000

  

  

    
 

    
rs55882921

    
C/T

    
CC:1.0000

      
CT: 0.0000

      
TT:0.0000

    
CC:1.0000

      
CT: 0.0000  TT:0.0000

  

  

    
 

    
rs41275697

    
C/T

    
CC:1.0000

      
CT: 0.0000

      
TT:0.0000

    
CC:1.0000

      
CT: 0.0000  TT:0.0000

  

  

    
 

    
rs1126440

    
C/T

    
CC:0.0000

      
CT: 0.0000

      
TT:1.0000

    
CC:0.0000

      
CT: 0.0000  TT:1.0000

  

  

    
 

    
rs41275699

    
A/G

    
AA:1.0000

      
AG:0.0000

      
GG:0.0000

    
AA:1.0000  AG:0.0000

      
GG:0.0000

  

  

    
 

    
rs67420531

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
ADH3

    
rs56247447

    
C/T

    
CC:0.9900

      
CT:0.0100

      
TT:0.0000

    
CC:1.0000  CT:0.0000

      
TT:0.0000

  

  

    
 

    
rs698

    
A/G

    
AA:0.8400

      
AG:0.1500

      
GG:0.0100

    
AA:0.8889  AG:0.1111

      
GG:0.0000

  

  

    
 

    
rs1042756

    
C/G

    
CC:1.0000

      
CG:0.0000

      
GG:0.0000

    
CC:1.0000  CG:0.0000

      
GG:0.0000

  

  

    
 

    
rs55717907

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs2241894

    
A/G

    
AA:0.0400

      
AG:0.9600

      
GG:0.0000

    
AA:0.0505  AG:0.9495

      
GG:0.0000

  

  

    
 

    
rs1789915

    
A/G

    
AA:0.9800

      
AG:0.0200

      
GG:0.0000

    
AA:0.9899  AG:0.0101

      
GG:0.0000

  

  

    
 

    
rs6490301

    
C/T

    
CC:1.0000

      
CT:0.0000

      
TT:0.0000

    
CC:1.0000  CT:0.0000

      
TT:0.0000

  

  

    
ALDH2

    
rs1064903

    
C/G

    
CC:0.0000

      
CG:0.0000

      
GG:1.0000

    
CC:0.0000  CG:0.0000

      
GG:1.0000

  

  

    
 

    
rs13306164

    
C/T

    
CC:0.9200

      
CT:0.0800

      
TT:0.0000

    
CC:0.8788  CT:0.1212

      
TT:0.0000

  

  

    
 

    
rs58280059

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs1062136

    
A/T

    
AA:1.0000

      
AT:0.0000

      
TT:0.0000

    
AA:1.0000  AT:0.0000

      
TT:0.0000

  

  

    
 

    
rs1064933

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs671

    
A/G

    
AA:0.0000

      
AG:0.1000

      
GG:0.9000

    
AA:0.0404  AG:0.4141

      
GG:0.5455

  

  

    
 

    
rs59868347

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
CYP2E1

    
rs35844228

    
C/G

    
CC:0.0000

      
CG:0.0000

      
GG:1.0000

    
CC:0.0000  CG:0.0000

      
GG:1.0000

  

  

    
 

    
rs72559710

    
A/C/G

    
AG:0.0100

      
CG:0.0000

      
GG:0.9900

    
AG:0.0101  CG:0.0101  GG:0.9798

  

  

    
 

    
rs28371740

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs60719153

    
C/T

    
CC:1.0000

      
CT:0.0000

      
TT:0.0000

    
CC:1.0000  CT:0.0000

      
TT:0.0000

  

  

    
 

    
rs56864127

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs60452492

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs56040284

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs28371743

    
C/T

    
CC:0.0000

      
CT:0.0000

      
TT:1.0000

    
CC:0.0000

      
CT:0.0000

      
TT:1.0000

  

  

    
 

    
rs41299426

    
A/G

    
AA:1.0000

      
AG:0.0000

      
GG:0.0000

    
AA:1.0000  AG:0.0000

      
GG:0.0000

  

  

    
 

    
rs61710826

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs28371746

    
A/C

    
AA:0.0000

      
AC:0.0200

      
CC:0.9800

    
AA:0.0000

      
AC:0.0606

      
CC:0.9394

  

  

    
 

    
rs61644766

    
C/G

    
CC:1.0000

      
CG:0.0000

      
GG:0.0000

    
CC:1.0000  CG:0.0000

      
GG:0.0000

  

  

    
 

    
rs41299434

    
C/G

    
CC:1.0000

      
CG:0.0000

      
GG:0.0000

    
CC:1.0000  CG:0.0000

      
GG:0.0000

  

  

    
 

    
rs59981143 

    
A/G

    
AA:0.9900

      
AG:0.0100

      
GG:0.0000

    
AA:1.0000  AG:0.0000

      
GG:0.0000

  

  

    
 

    
rs55897648

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  

  

    
 

    
rs60207639

    
A/G

    
AA:1.0000

      
AG:0.0000

      
GG:0.0000 

    
AA:1.0000  AG:0.0000

      
GG:0.0000

  

  

    
 

    
rs59656378

    
C/G

    
CC:0.0000

      
CG:0.0000

      
GG:1.0000

    
CC:0.0000  CG:0.0000

      
GG:1.0000

  

  

    
 

    
rs57702102

    
A/G

    
AA:1.0000

      
AG:0.0000

      
GG:0.0000

    
AA:1.0000  AG:0.0000

      
GG:0.0000

  

  

    
 

    
rs2515641

    
C/T

    
CC:0.6200

      
CT:0.3600

      
TT:0.0200

    
CC:0.6566  CT:0.3333

      
TT:0.0101

  

  

    
 

    
rs55982231

    
A/G

    
AA:0.0000

      
AG:0.0000

      
GG:1.0000

    
AA:0.0000  AG:0.0000

      
GG:1.0000

  










Table 3:  Genotype frequencies of 40 SNPs in intoxicated population (n=100) and control population (n=99).

Table 4: Allele frequencies of seven polymorphic SNPs in intoxicated population (n=100) and control population (n=99).





  

    
Gene

    
SNP Loci

    
Variation

    
Allele frequencies in intoxicated population

    
Allele frequencies in control population

  

  

    
ADH3

    
rs698

    
A/G

    
0.9150/0.0850

    
0.9444/0.0556

  

  

    
 

    
rs2241894

    
A/G

    
0.5200/0.4800

    
0.5253/0.4747

  

  

    
 

    
rs1789915

    
A/G

    
0.9900/0.0100

    
0.9949/0.0051

  

  

    
ALDH2

    
rs13306164

    
C/T

    
0.9600/0.0400

    
0.9394/0.0606

  

  

    
 

    
rs671

    
A/G

    
0.0500/0.9500

    
0.2475/0.7525

  

  

    
CYP2E1

    
rs28371746

    
A/C

    
0.0100/0.9900

    
0.0303/0.9697

  

  

    
 

    
rs2515641

    
C/T

    
0.8000/0.2000

    
0.8232/0.1768

  










Table 4:  Allele frequencies of seven polymorphic SNPs in intoxicated population (n=100) and control population (n=99).

Population comparison

The allele frequencies of seven polymorphic SNPs (rs698, rs2241894, rs1789915, rs13306164, rs671, rs28371746, rs2515641) in intoxicated population were compared to the data about the control population. It’s found that two SNPs (rs671 in and rs2515641) with a significant difference in frequency distribution between intoxicated population and control population (p<0.01, (Table 5) might be related to alcohol intoxication.

Table 5: Comparison of allele frequency of seven polymorphic SNPs in intoxicated population (n=100) and control population (n=99).





  

    
Population

    
Numbers of allele 1

    
Numbers of allele 2

    
Total of alleles

    
X2

    
P value

  

  

    
rs698（A/G） 

  

  

    
intoxicated population

    
183

    
17

    
200

    
1.3189

    
0.2508

  

  

    
control population

    
187

    
11

    
198

    
 

    
 

  

  

    
rs2241894(A/G)

  

  

    
intoxicated population

    
104

    
96

    
200

    
0.0110

    
0.9165

  

  

    
control population

    
104

    
94

    
198

    
 

    
 

  

  

    
rs1789915(A/G)

  

  

    
intoxicated population

    
198

    
2

    
200

    
0.0000

    
1.0000

  

  

    
control population

    
197

    
1

    
198

    
 

    
 

  

  

    
rs13306164(C/T)

  

  

    
intoxicated population

    
192

    
8

    
200

    
0.8852

    
0.3468

  

  

    
control population

    
186

    
12

    
198

    
 

    
 

  

  

    
rs671(A/G)

  

  

    
intoxicated population

    
10

    
190

    
200

    
30.7291

    
<0.0001

  

  

    
control population

    
49

    
149

    
198

    
 

    
 

  

  

    
rs28371746(A/C)

  

  

    
intoxicated population

    
2

    
198

    
200

    
1.1791

    
0.2775

  

  

    
control population

    
6

    
192

    
198

    
 

    
 

  

  

    
rs2515641(C/T)

  

  

    
intoxicated population

    
160

    
40

    
200

    
4.6574

    
0.0309

  

  

    
control population

    
163

    
35

    
198

    
 

    
 

  










Table 5:  Comparison of allele frequency of seven polymorphic SNPs in intoxicated population (n=100) and control population (n=99).

References

  1. Sim SC, Risinger C, Dahl ML, Aklillu E, Christensen M, Bertilsson L, et al. A common novel CYP2C19 gene variant causes ultrarapid drug metabolism relevant for the drug response to proton pump inhibitors and antidepressants. Clin Pharmacol Ther. 2006; 79: 103-113.
  2. Meyer UA, Zanger UM. Molecular mechanisms of genetic polymorphisms of drug metabolism. Annu Rev Pharmacol Toxicol. 1997; 37: 269-296.
  3. Shi Y, Xiang P, Li L, Shen M. Analysis of 50 SNPs in CYP2D6, CYP2C19, CYP2C9, CYP3A4 and CYP1A2 by MALDI-TOF mass spectrometry in Chinese Han population. Forensic Sci Int. 2011; 207: 183-187.
  4. Zhong Y,Cao J, Zou R, Peng M. Genetic polymorphisms in alcohol dehydrogenase, aldehyde dehydrogenase and alcoholic chronic pancreatitis susceptibility: A meta-analysis, 2014; S0210-5705: 00304-5.
  5. Wall TL. Genetic associations of alcohol and aldehyde dehydrogenase with alcohol dependence and their mechanisms of action. Ther Drug Monit. 2005; 27: 700-703.
  6. Chai YG, Oh DY, Chung EK, Kim GS, Kim L, Lee YS, et al. Alcohol and aldehyde dehydrogenase polymorphisms in men with type I and Type II alcoholism. Am J Psychiatry. 2005; 162: 1003-1005.
  7. Toselli F, Booth Depaz IM, Worrall S, Etheridge N, Dodd PR, Wilce PA, et al. Expression of CYP2E1 and CYP2U1 proteins in amygdala and prefrontal cortex: influence of alcoholism and smoking. Alcohol Clin Exp Res. 2015; 39: 790-797.
  8. Yokoyama A, Mizukami T, Matsui T, Yokoyama T, Kimura M, Matsushita S, et al. Genetic polymorphisms of alcohol dehydrogenase-1B and aldehyde dehydrogenase-2 and liver cirrhosis, chronic calcific pancreatitis, diabetes mellitus, and hypertension among Japanese alcoholic men. Alcohol Clin Exp Res. 2013; 37: 1391-1401.
  9. Iyer-Eimerbrink PA, Nurnberger JI Jr. Genetics of alcoholism. Curr Psychiatry Rep. 2014; 16: 518.
  10. Petkovski E, Keyser-Tracqui C, Hienne R, Ludes B. SNPs and MALDI-TOF MS: tools for DNA typing in forensic paternity testing and anthropology. J Forensic Sci. 2005; 50: 535-541.
  11. Tost J, Gut IG. Genotyping single nucleotide polymorphisms by MALDI mass spectrometry in clinical applications. Clin Biochem. 2005; 38: 335-350.

Citation: Li L, Xiang P, Liu Y, Shi Y, Lin Y and Zhao Z. Comparison of SNP Polymorphism of Alcohol Metabolizing Related Enzyme Genes in Intoxicated Individuals and Nonalcoholic Population. Austin J Forensic Sci Criminol. 2015; 2(3): 1031. ISSN : 2380-0801