UJ 35 Summer.pdf


541Vol. 9   |   No. 3   |   Summer 2012   |U R O LO G Y   J O U R N A L

Purpose: To determine the role of glutathione S-transferases (GSTs; GSTM1, GSTT1, and GSTP1) 
gene polymorphisms in susceptibility to male factor infertility.

Materials and Methods: We report a pooled analysis of 11 studies on the association of GSTM1, 
GSTT1, and GSTP1 polymorphisms and male factor infertility, including 1323 cases and 1054 
controls. 

Results:
P = .003], GSTT1 null genotype 

P

P = .26). 

Conclusion: These results demonstrated that amongst populations studied to date, GSTM1 and 

Keywords: glutathione S-transferase, single-nucleotide polymorphisms, infertility, male, genetic, 

1Clinical Center for Urolog-
ical Disease Diagnosis and 
Private Clinic Specialized in 
Urological and Andrologi-
cal Genetics, Tehran, Iran
2Department of Urology, 
Shahid Modarres Hospital, 
Shahid Beheshti University 
of Medical Sciences, Teh-
ran, Iran
3Department of Health 
and Community Medicine, 
Shahid Beheshti University 
of Medical Sciences, Teh-
ran, Iran

Mohammad Reza Safarinejad,1 Farid Dadkhah,2 Majid Ali Asgari,2 Seyed Yousef Hosseini,2

Ali Asgar Kolahi,3 Elham Iran-Pour1

REVIEW

Glutathione S–Transferase Polymor-
phisms (GSTM1, GSTT1, GSTP1) and Male 
Factor Infertility Risk
A Pooled Analysis of Studies

Corresponding Author:

Mohammad Reza Safa-
rinejad, MD
P.O. Box 19395-1849, 
Tehran, Iran

Tel: +98 21 2245 4499
Fax: +98 21 2245 6845
E-mail: info@safarinejad.
com

Received July 2012
Accepted July 2012



542 | Review

INTRODUCTION

Wfrom primary or secondary infertility.(1) The cause of infertility is still not determined in 
infertility.(2)

(3) -
plicated.(4)

number of male infertility is due to gene mutations and 
single-nucleotide polymorphisms (SNP).
An increasing number of studies are determining the as-

(9-12)

implication. By determining the underlying genetic basis 

-
ity and genetic polymorphisms of GST genes. 
Gene and Gene Function

a superfamily of multifunctional enzymes that detoxify 
products of oxidative stress, environmental substances, 
and reactive electrophiles.(13,14) Glutathione S-transferases 
also inactivate carcinogens by catalyzing the conjugation 
of electrophiles to glutathione.(15) DNA damage due to en-
dogenously formed lipid peroxidase can be prevented by 
GSTs.(16)

GST isoenzymes have been assigned to eight separate 
classes, including  (Alpha), μ (Mu),  (Ome-
ga),  (Pi),  (Sigma),  (Theta), and 
encoded by the GSTA, GSTM, GSTK, GSTO, GSTP, 
GSTS, GSTT, and GSTZ genes, respectively. -
dition, each class includes several genes and isoenzymes.
(19) Polymorphisms have been reported in the GSTM1, 
GSTT1, and GSTP1 genes coding for GSTs enzymes in 
the Mu, Theta, and Pi classes, respectively.

on chromosome 1p13.3.(20) GSTM1 has a common func-
tional variant (null versus present). The frequency of this 

-
ulation studied.(21)

of the GSTM1 locus have no enzymatic functional activity 
of the cytosolic enzyme GST-μ.(22) The homozygous dele-

(23-25) 

The human Theta class of GSTs (GSTT) is comprised of 
-

cated on chromosome 22q11.(26) The polymorphism in the 
GSTT1 gene loci is also caused by a gene deletion and 
brings about in virtual absence of enzyme activity in per-

(27)

both GSTT1 null genotype(23,24) and nondeletion genotype 
of the GSTT1 gene has also been reported.

(29)

gene. A single-nucleotide polymorphism at position 313 in 
(30) The 

resulting isoleucine to valine substitution in codon 105 of 
105 105

activity.(31) Another SNP at codon 114 leads to alanine 
-

ference in catalytic activity.(32)

(23) But in a study 

(33) 

GST and Male Infertility Risk
Excessive reactive oxygen species (ROS) have been sug-
gested to be one of the major contributory factors resulting 
in male infertility via oxidative DNA damages.(34) Glu-
tathione S-transferase is one of the human defense mecha-
nisms opposing the deleterious effects of oxidative stress.
(35) Glutathione S-transferases gene polymorphisms could 
impair the capability of defense against oxidative stress 
and result in the development of some cancers.(36) One of 
the determinant factors of susceptibility of spermatozoa to 
oxidative damage is GSTM1 polymorphism.(25) DNA frag-
mentation in human sperm can be modulated by GSTM1 
gene polymorphism.(37)

Objective
There have been various studies in the literature regarding 
the association of GST (M1, T1, and P1) polymorphisms 

 (Table 1). There is sig-



543Vol. 9   |   No. 3   |   Summer 2012   |U R O LO G Y   J O U R N A L

GST Polymorphisms and Male Factor Infertility   |  Safarinejad et al

to determine the overall effect of GST (M1, T1, and P1) 

MATERIALS AND METHODS
Data Collection

-
ciation of GST polymorphisms (GSTM1, GSTT1, and 

the search terms of “GST OR Glutathione S-transferase 
AND polymorphism OR polymorphisms AND variant OR 
variants AND infertility OR infertile AND male”, yielding 
11 results. 
Statistical Analysis

using unconditional logistic regression model, including 

-
P -

ed using ORs from the individual included studies. To as-

-

-

RESULTS
The results of the pooled analysis are summarized in Ta-

-

P = .003), GSTT1 null genotype (OR, 
P = .02), and male infer-

-

Table 1. Summary of published studies on the association between GSTM1, GSTT1, and GSTP1 polymorphisms and male infertility.

First author (Ref no.) Publication year Cases Controls Country Ethnicity Studied gene(s)

Tang (33) 2012 65 30 China Asian GSTM1, GSTT1, GSTP1

Safarinejad(23) 2010 166 166 Iran Asian GSTM1, GSTT1, GSTP1

Tirumala(40) 2010 42 43 India Asian GSTM1

Polonikov(28) 2010 203 227 Russia Caucasian GSTM1, GSTT1

Ichioka(41) 2009 274 101 Japan Asian GSTM1, GSTT1

Wu(42) 2009 63 54 China Asian GSTT1

Finotti(24) 2009 128 105 Brazil Caucasian GSTM1, GSTT1

Aydos(38) 2009 110 105 Turkey Caucasian GSTM1

Wu(43) 2008 78 103 China Asian GSTT1

Aydemir(25) 2007 52 60 Turkey Caucasian GSTM1

Chen(39) 2002 142 60 China Asian GSTM1



544 | Review

P = 
.007) and GSTT1 null genotype (P = .02). No evidence of 

P
= .62) nor for the GSTM1 genotype (P = .24).
On the other hand, GSTP1 polymorphism had overall 
protective effect against development of male infertility, 

P

P

GSTP1 (P P
= .12). 
The sensitivity analysis demonstrated that exclusion of 

(33) -
sociates(41) from the analysis decreased the evidence of 
heterogeneity (GSTM1: P = .02; GSTT1: P = .04; and 

GSTP1: P = 1.0). After performing sensitivity analysis, 

null genotype (P P = 

null genotype (P = .04). 
-

-

-
P = 

Trend P

Table 2. Association between GSTM1 and male infertility in the
pooled analysis.

Genotype Adjusted Odds Ratio* Trend P

Overall .003

Present 1.00 (Ref )

Null 2.47 (1.72 to 3.84)

Never smoker .004

Present 1.00 (Ref )

Null 2.21 (1.52 to 3.57)

Ever smoker .002

Present 1.00 (Ref )

Null 2.88 (1.91 to 4.16)

Infertility .002

Present 1.00 (Ref )

Null 2.71 (1.84 to 3.92)

*Odds Ratios are adjusted for study, age, race, and smoking 
history.

Table 3. Association between GSTT1 and male infertility in the
pooled analysis.

Genotype Adjusted Odds Ratio* Trend P

Overall .02

Present 1.00 (Ref )

Null 1.54 (1.43 to 3.47)

Never smoker .03

Present 1.00 (Ref )

Null 1.36 (1.57 to 3.12)

Ever smoker .01

Present 1.00 (Ref )

Null 1.72 (1.63 to 3.74)

Infertility .01

Present 1.00 (Ref )

Null 1.68 (1.57 to 3.69)

*Odds Ratios are adjusted for study, age, race, and smoking 
history.



545Vol. 9   |   No. 3   |   Summer 2012   |U R O LO G Y   J O U R N A L

to 3.12; Trend P
3.74; Trend P -

-
types in both the analysis restricted to the homogeneous 
studies and in the full-pooled analysis. 

DISCUSSION
This pooled analysis, containing 1054 controls and 1323 
cases from 11 studies, demonstrates an overall association 

-

-

and GSTM1 and GSTT1 null genotypes. The direction of 
the interaction is the same to the expected effect.
Strength of our pooled analysis is that the collection of 

-

has advantages over meta-analysis.(44) Meta-analysis al-
-

-
-
-

(45) 

(46) and being 
(47) 

-
-

Asian populations(23,33,39-43)

Table 4. Association between GSTP1 and male infertility in the
pooled analysis.

Genotype Adjusted Odds Ratio* Trend P

Overall .002

Ile/Ile 1.00 (Ref )

Ile/Val 0.48 (0.27 to 0.77) 

Val/Val 1.81 (0.34 to 7.85)

Ile/Val or Val/Val 0.52 (0.31 to 0.81)

Never smoker .03

Ile/Ile 1.00 (Ref )

Ile/Val 0.52 (0.33 to 0.80)

Val/Val 1.84 (0.37 to 7.42)

Ile/Val or Val/Val 0.58 (0.38 to 0.87)

Ever smoker .01

Ile/Ile 1.00 (Ref )

Ile/Val 0.50 (0.31 to 0.79)

Val/Val 1.82 (0.35 to 37.66)

Ile/Val or Val/Val 0.54 (0.34 to 0.84)

Infertility .01

Ile/Ile 1.00 (Ref )

Ile/Val 0.54 (0.37 to 0.81)

Val/Val 1.80 (0.41 to 7.12)

Ile/Val or Val/Val 0.57 (0.36 to 0.86)

* Odds Ratios are adjusted for study, age, race, and smoking 
history.

GST Polymorphisms and Male Factor Infertility   |  Safarinejad et al



546 |

-

GSTP1 polymorphisms and male infertility in ethnicities 

genotypes and male infertility.

CONCLUSION

and GSTP1 polymorphisms and male factor infertility, and 
-

CONFLICT OF INTEREST
None declared.

Review

REFERENCES 

1. World Health Organization. In: Vayena E, Rowe PJ, Griffin 
PD, eds. Medical, Ethical and Social Aspects of Assisted Repro-
duction (2001, Geneva), Current Practices and Controversies 
in Assisted Reproduction: Report of a WHO Meeting. Geneva, 
Switzerland: WHO Publications; 2002.

2. Nieschlag E. Scope and goals of andrology. In: Nieschlag 
E, Behre HM, eds. Andrology: Male Reproductive Health and 
Dysfunction. 2 ed. Heidelberg: Springer-Verlag; 2000:1-8.

3. Safarinejad MR. Infertility among couples in a population-
based study in Iran: prevalence and associated risk factors. 
Int J Androl. 2008;31:303-14.

4. Brugh VM, 3rd, Lipshultz LI. Male factor infertility: evalua-
tion and management. Med Clin North Am. 2004;88:367-
85.

5. Matzuk MM, Lamb DJ. The biology of infertility: research 
advances and clinical challenges. Nat Med. 2008;14:1197-
213.

6. Safarinejad MR, Shafiei N, Safarinejad S. The role of en-
dothelial nitric oxide synthase (eNOS) T-786C, G894T, and 
4a/b gene polymorphisms in the risk of idiopathic male 
infertility. Mol Reprod Dev. 2010;77:720-7.

7. O'Flynn O'Brien KL, Varghese AC, Agarwal A. The genetic 
causes of male factor infertility: a review. Fertil Steril. 
2010;93:1-12.

8. Safarinejad MR, Shafiei N, Safarinejad S. Association of 
polymorphisms in the estrogen receptors alpha, and beta 
(ESR1, ESR2) with the occurrence of male infertility and se-
men parameters. J Steroid Biochem Mol Biol. 2010;122:193-
203.

9. Vicdan A, Vicdan K, Gunalp S, et al. Genetic aspects of 
human male infertility: the frequency of chromosomal 
abnormalities and Y chromosome microdeletions in severe 
male factor infertility. Eur J Obstet Gynecol Reprod Biol. 
2004;117:49-54.

10. Safarinejad MR, Shafiei N, Safarinejad S. Association of the 
(TAAAA)n repeat and Asp327Asn polymorphisms in the sex 
hormone-binding globulin (SHBG) gene with idiopathic 
male infertility and relation to serum SHBG concentrations. 
J Steroid Biochem Mol Biol. 2011;123:37-45.

11. Maduro MR, Lamb DJ. Understanding new genetics of 
male infertility. J Urol. 2002;168:2197-205.

12. Safarinejad MR, Shafiei N, Safarinejad S. Relationship 
between genetic polymorphisms of methylenetetrahydro-
folate reductase (C677T, A1298C, and G1793A) as risk factors 
for idiopathic male infertility. Reprod Sci. 2011;18:304-15.

13. Sanyal S, Festa F, Sakano S, et al. Polymorphisms in DNA 
repair and metabolic genes in bladder cancer. Carcinogen-
esis. 2004;25:729-34.

14. Schnakenberg E, Breuer R, Werdin R, Dreikorn K, Schloot W. 
Susceptibility genes: GSTM1 and GSTM3 as genetic risk fac-
tors in bladder cancer. Cytogenet Cell Genet. 2000;91:234-
8.

15. Hayes JD, Pulford DJ. The glutathione S-transferase super-
gene family: regulation of GST and the contribution of the 
isoenzymes to cancer chemoprotection and drug resist-
ance. Crit Rev Biochem Mol Biol. 1995;30:445-600.

16. Hurst R, Bao Y, Jemth P, Mannervik B, Williamson G. Phos-
pholipid hydroperoxide glutathione peroxidase activity of 
human glutathione transferases. Biochem J. 1998;332 ( Pt 
1):97-100.

17. Lo HW, Ali-Osman F. Genetic polymorphism and function 
of glutathione S-transferases in tumor drug resistance. Curr 
Opin Pharmacol. 2007;7:367-74.

18. Strange RC, Spiteri MA, Ramachandran S, Fryer AA. 
Glutathione-S-transferase family of enzymes. Mutat Res. 
2001;482:21-6.

19. Hayes JD, Strange RC. Glutathione S-transferase polymor-
phisms and their biological consequences. Pharmacology. 
2000;61:154-66.



547Vol. 9   |   No. 3   |   Summer 2012   |U R O LO G Y   J O U R N A L

20. Rebbeck TR. Molecular epidemiology of the human 
glutathione S-transferase genotypes GSTM1 and GSTT1 in 
cancer susceptibility. Cancer Epidemiol Biomarkers Prev. 
1997;6:733-43.

21. Garte S, Gaspari L, Alexandrie AK, et al. Metabolic gene 
polymorphism frequencies in control populations. Cancer 
Epidemiol Biomarkers Prev. 2001;10:1239-48.

22. Zhong S, Howie AF, Ketterer B, et al. Glutathione S-trans-
ferase mu locus: use of genotyping and phenotyping 
assays to assess association with lung cancer susceptibility. 
Carcinogenesis. 1991;12:1533-7.

23. Safarinejad MR, Shafiei N, Safarinejad S. The association of 
glutathione-S-transferase gene polymorphisms (GSTM1, 
GSTT1, GSTP1) with idiopathic male infertility. J Hum 
Genet. 2010;55:565-70.

24. Finotti AC, Costa ESRC, Bordin BM, Silva CT, Moura KK. 
Glutathione S-transferase M1 and T1 polymorphism in men 
with idiopathic infertility. Genet Mol Res. 2009;8:1093-8.

25. Aydemir B, Onaran I, Kiziler AR, Alici B, Akyolcu MC. In-
creased oxidative damage of sperm and seminal plasma 
in men with idiopathic infertility is higher in patients with 
glutathione S-transferase Mu-1 null genotype. Asian J 
Androl. 2007;9:108-15.

26. Tan KL, Webb GC, Baker RT, Board PG. Molecular cloning of 
a cDNA and chromosomal localization of a human theta-
class glutathione S-transferase gene (GSTT2) to chromo-
some 22. Genomics. 1995;25:381-7.

27. Pemble S, Schroeder KR, Spencer SR, et al. Human glu-
tathione S-transferase theta (GSTT1): cDNA cloning and the 
characterization of a genetic polymorphism. Biochem J. 
1994;300 ( Pt 1):271-6.

28. Polonikov AV, Yarosh SL, Kokhtenko EV, Starodubova NI, 
Pakhomov SP, Orlova VS. The functional genotype of glu-
tathione S-transferase T1 gene is strongly associated with 
increased risk of idiopathic infertility in Russian men. Fertil 
Steril. 2010;94:1144-7.

29. Kano T, Sakai M, Muramatsu M. Structure and expression of 
a human class pi glutathione S-transferase messenger RNA. 
Cancer Res. 1987;47:5626-30.

30. Ahmad H, Wilson DE, Fritz RR, et al. Primary and secondary 
structural analyses of glutathione S-transferase pi from hu-
man placenta. Arch Biochem Biophys. 1990;278:398-408.

31. Watson MA, Stewart RK, Smith GB, Massey TE, Bell DA. 
Human glutathione S-transferase P1 polymorphisms: 
relationship to lung tissue enzyme activity and population 
frequency distribution. Carcinogenesis. 1998;19:275-80.

32. Ali-Osman F, Akande O, Antoun G, Mao JX, Buolamwini 
J. Molecular cloning, characterization, and expression 
in Escherichia coli of full-length cDNAs of three human 
glutathione S-transferase Pi gene variants. Evidence for 
differential catalytic activity of the encoded proteins. J Biol 
Chem. 1997;272:10004-12.

33. Tang K, Xue W, Xing Y, et al. Genetic polymorphisms of glu-
tathione S-transferase M1, T1, and P1, and the assessment 
of oxidative damage in infertile men with varicoceles from 
northwestern China. J Androl. 2012;33:257-63.

34. Saleh RA, Agarwal A. Oxidative stress and male infertil-
ity: from research bench to clinical practice. J Androl. 
2002;23:737-52.

35. Awasthi YC, Sharma R, Singhal SS. Human glutathione S-
transferases. Int J Biochem. 1994;26:295-308.

36. Ryberg D, Skaug V, Hewer A, et al. Genotypes of glu-
tathione transferase M1 and P1 and their significance for 
lung DNA adduct levels and cancer risk. Carcinogenesis. 
1997;18:1285-9.

37. Rubes J, Selevan SG, Sram RJ, Evenson DP, Perreault SD. 
GSTM1 genotype influences the susceptibility of men to 
sperm DNA damage associated with exposure to air pollu-
tion. Mutat Res. 2007;625:20-8.

38. Aydos SE, Taspinar M, Sunguroglu A, Aydos K. Association 
of CYP1A1 and glutathione S-transferase polymorphisms 
with male factor infertility. Fertil Steril. 2009;92:541-7.

39. Chen SS, Chang LS, Chen HW, Wei YH. Polymorphisms of 
glutathione S-transferase M1 and male infertility in Taiwan-
ese patients with varicocele. Hum Reprod. 2002;17:718-25.

40. Tirumala Vani G, Mukesh N, Siva Prasad B, et al. Role of 
glutathione S-transferase Mu-1 (GSTM1) polymorphism in 
oligospermic infertile males. Andrologia. 2010;42:213-7.

41. Ichioka K, Nagahama K, Okubo K, Soda T, Ogawa O, Nishiy-
ama H. Genetic polymorphisms in glutathione S-trans-
ferase T1 affect the surgical outcome of varicocelectomies 
in infertile patients. Asian J Androl. 2009;11:333-41.

42. Wu Q, Xing J, Xue W, Sun J, Wang X, Jin X. Influence of 
polymorphism of glutathione S-transferase T1 on Chinese 
infertile patients with varicocele. Fertil Steril. 2009;91:960-2.

43. Wu QF, Xing JP, Tang KF, et al. Genetic polymorphism of 
glutathione S-transferase T1 gene and susceptibility to 
idiopathic azoospermia or oligospermia in northwestern 
China. Asian J Androl. 2008;10:266-70.

44. Blettner M, Sauerbrei W, Schlehofer B, Scheuchenpflug 
T, Friedenreich C. Traditional reviews, meta-analyses and 
pooled analyses in epidemiology. Int J Epidemiol. 1999;28:1-
9.

GST Polymorphisms and Male Factor Infertility   |  Safarinejad et al



548 |

45. Friedenreich CM. Commentary: improving pooled analyses 
in epidemiology. Int J Epidemiol. 2002;31:86-7.

46. Kusters M, Valentini V, Calvo FA, et al. Results of European 
pooled analysis of IORT-containing multimodality treatment 
for locally advanced rectal cancer: adjuvant chemotherapy 
prevents local recurrence rather than distant metastases. 
Ann Oncol. 2010;21:1279-84.

47. Chen J, Germer S, Higuchi R, Berkowitz G, Godbold J, Wet-
mur JG. Kinetic polymerase chain reaction on pooled DNA: 
a high-throughput, high-efficiency alternative in genetic 
epidemiological studies. Cancer Epidemiol Biomarkers Prev. 
2002;11:131-6.

Review