Increased oxidative damage of sperm and seminal plasma in men with idiopathic infertility is higher in patients with glutathione S-transferase Mu-1 null genotype

Aim： To examine whether a relationship exists between glutathione S-transferase Mu-1 （GSTM1） gene polymorphism and the susceptibility of sperm and seminal plasma from patients with idiopathic infertility to oxidative stress. Methods： Fifty-two men with idiopathic infertility and 60 healthy fertile men were recruited to this study. GSTM1 gene polymorphism was determined by polymerase chain reaction （PCR） and both the infertile and control individuals were divided into GSTM1 null and GSTM1 positive groups according to their GSTM1 gene structure. We compared reactive oxygen species （ROS） generation, malondialdehyde （MDA）, protein carbonyls and glutathione （GSH） concentrations, and glutathione S-transferase （GST） activity in seminal plasma and spermatozoa from infertile patients and controls with respect to GSTM1 genotype. Results： Significantly higher levels of oxidative stress and damage markers were found in idiopathic infertile men with the GSTM1 null genotype compared with those with the GSTM1 positive genotype. There was no significant difference in genotype distribution for theGSTM1 variant between the idiopathic infertile subjects and fertile subjects. Patients with the GSTM1 null genotype also had lower sperm concentrations than those with GSTM1 positive genotype. Conclusion： Our results suggest that the susceptibility of sperm and seminal plasma to oxidative stress is significantly greater in idiopathic infertile men with the GSTM1 null genotype compared with those possessing the gene. Therefore, in patients with idiopathic infertility, GSTM1 polymorphism might be an important source of variation in susceptibility of spermatozoa to oxidative damage.

Genetic polymorphism of glutathione S-transferase T1 gene and susceptibility to idiopathic azoospermia or oligospermia in northwestern China

Aim： To investigate the association of glutathione S-transferase T1 （GSTT1） gene polymorphism in patients with idiopathic azoospermia or oligospermia in the northwestern China population. Methods： In the case-control study, GSTT1 genotypes were identified by multiplex polymerase chain reaction （PCR） with peripheral blood DNA samples from 78 patients with idiopathic azoospermia, 103 patients with idiopathic oligospermia and 156 age-matched controls with normal sperm concentration and motility, according to the criteria adapted from World Health Organization guidelines. All of the patients and controls were from northwestern China. Results： There is a significant association between GSTT1 null genotype with idiopathic azoospermia risk （odds ratio [OR]： 2.36, 95% confidence interval [CI]： 1.33-4.20, P = 0.003） or idiopathic oligospermia risk （OR： 2.00, 95% CI： 1.17-3.27, P = 0.010）. Conclusion： GSTT1 null genotype is a predisposing risk factor for sporadic idiopathic azoospermia or oligospermia in northwestern China. （Asian J Androl 2008 Mar; 10： 266-270）

Anticancer Drug Resistance of HeLa Cells Transfected With Rat Glutathione S-transferase pi Gene

To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe. Results HeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 靏/mL, 10.95 靏/mL and 16.52 靏/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 靏/mL, 7.48 靏/mL and 13.70 靏/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different. Conclusions Our findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research.

Association of glutathione S-transferase T1 and M1 gene polymorphisms with ischemic stroke risk in the Chinese Han population

Atherosclerosis plays an important role in ischemic stroke, and oxidative stress participates in the entire process of atherosclerosis. Glutathione S-transferase （GST） acting with other antioxidant enzymes can eliminate reactive oxygen species and protect cells against oxidative damage. To assess the association of glutathione S-transferase （GSTT1 and GSTM1） gene polymorphisms with ischemic stroke in the Chinese Han population, the present study selected 315 patients with ischemic stroke and 210 healthy controls for comparison. GSTT1 and GSTM1 genotypes were determined using polymerase chain reactions, electrophoresis and imaging analysis. No obvious evidence of GSTTI-nulI, GSTMI-null and GSTTI/GSTMI-double null genotype distribution differences was found between case and control groups or between genders. Subgroup analysis showed that the risk of stroke was increased when hypertension was accompanied by GSTTl-null （odds ratio （OR） = 2.996, P 〈 0.001） and GSTMl-null （OR = 3.680, P 〈 0.001 ） genotypes; diabetes mellitus was accompanied by GSTTI-null （OR = 1.860, P = 0.031） and GSTMI-null （OR = 2.444, P = 0.002） genotypes, and smokers showed a GSTTl-null genotype （OR = 2.276, P = 0.003）. GSTT1- and GSTMl-null genotypes may interact synergistically with hypertension, diabetes mellitus and smoking to increase the incidence risk of ischemic stroke.

A systemic review of glutathione S-transferase P1 Ile105Val polymorphism and colorectal cancer risk

Objectives： To investigate the correlation between glutathione S-transferase P1 （GSTP1） Ilel05Val polymorphism and colorectal cancer （CRC） risk. Methods： Studies were identified to investigate the association between GSTP1 Ilel05Val polymorphism and CRC risk. Systematic computerized searches of the PubMed, Chinese National Knowledge Infrastructure, WANFANG and SinoMed were performed. Summary odds ratios （OR） and 95% confidence intervals （95 % CI） were used to measure GSTP 1 Ile 105Val polymorphisms and CRC risk. Results： A total of 23 retrospective studies were included in the meta-analysis. During all studies including 6,981 cases and 8,977 controls, sample sizes ranged from 146 to 2,144. Overall, the pooled results revealed that lie 105Val polymorphism was not associated with CRC risk and confused results were found in subgroup analyses. Further meta-analyses were conducted after excluding low-quality studies. GSTP1 Ilel05Val is associated with increased risk of CRC limited in studies with matched control. There was no significant heterogeneity in all genetic comparisons, but heterogeneity existed in subgroup analyses of heterozygous and dominant comparisons. The meta-regression analyses indicated that matched controls were the significant factor influencing between-study heterogeneity in all possible influential factors including published year, ethnicity, source of control, sample size, Hardy-Weinberg equilibrium （HWE） in control and matched controls. Sensitivity analysis revealed the pooled ORs were not changed before and after removal of each single study in all genetic comparisons, indicating the robustness of the results. Conclusions： GSTP1 Ilel05Val might be associated with increased risk of CRC. However, more high- quality case-control studies should be performed to confirm the authenticity of our conclusion.

Rethinking de novo immune hepatitis,an old concept for liver allograft rejection:relevance of glutathione S-transferase T1 mismatch

Antibody-mediated rejection(AMR) in liver transplantation has long been underestimated. The concept of the liver as an organ susceptible to AMR has emerged in recent years, not only in the context of the major histocompatibility complex with the presence of HLA donor-specific antibodies, but also with antigens regarded as "minor", whose role in AMR has been demonstrated. Among them, antibodies against glutathione S-transferase T1 have been found in 100% of patients with de novo autoimmune hepatitis(dn AIH) when studied. In its latest update, the Banff Working Group for liver allograft pathology proposed replacing the term dn AIH with plasma cell(PC)-rich rejection. Antibodies to glutathione S-transferase T1(GSTT1) in null recipients of GSTT1 positive donors have been included as a contributory but nonessential feature of the diagnosis of PC-rich rejection. Also in this update, non-organ-specific anti-nuclear or smooth muscle autoantibodies are no longer included as diagnostic criteria. Although initially found in a proportion of patients with PC-rich rejection, the presence of autoantibodies is misleading since they are not diseasespecific and appear in many different contexts as bystanders. The cellular types and proportions of the inflammatory infiltrates in diagnostic biopsies have been studied in detail very recently. PC-rich rejection biopsies present a characteristic cellular profile with a predominance of T lymphocytes and a high proportion of PCs, close to 30%, of which 16.48% are Ig G4+. New data on the relevance of GSTT1-specific T lymphocytes to PC-rich rejection will be discussed in this review.

T-cell allorecognition of donor glutathione S-transferase T1 in plasma cell-rich rejection

AIM To investigate the role of glutathione S-transferase T1 donor-specific T lymphocytes in plasma cell-rich rejection of liver allografts.METHODS The study group included 22 liver transplant patients. Among them, 18 patients were mismatched for the glutathione S-transferase T1(GSTT1) alleles(don+/rec-), and 4 were matched(don+/rec+). Seven of the mismatched patients produced anti-GSTT1 antibodies and developed plasma cell-rich rejection(former de novo immune hepatitis). For the detection of specific Tlymphocytes, peripheral blood mononuclear cells were collected and stored in liquid nitrogen. The memory T cell response was studied by adding to the cell cultures to a mix of 39 custom-made, 15-mer overlapping peptides, which covered the entire GSTT1 amino acid sequence. The specific cellular response to peptides was analyzed by flow cytometry using the markers CD8, CD4, IL-4 and IFNγ.RESULTS Activation of CD8^+ T cells with different peptides was observed exclusively in the group of patients with plasma-cell rich rejection(3 out of 7), with production of IL-4 and/or IFNγ at a rate of 1%-4.92% depending on the peptides. The CD4^+ response was most common and not exclusive for patients with the disease, where 5 out of 7 showed percentages of activated cells from 1.24% to 31.34%. Additionally, two patients without the disease but with the mismatch had cells that became stimulated with some peptides(1.45%-5.18%). Highly unexpected was the finding of a double positive CD4^+CD8^(low) T cell population that showed the highest degree of activation with some of the peptides in 7 patients with the mismatch, in 4 patients with plasma cell-rich rejection and in 3 patients without the disease. Unfortunately, CD4^+CD8^(low) cells represent 1% of the total number of lymphocytes, and stimulation could not be analyzed in 9 patients due to the low number of gated cells. Cells from the 4 patients included as controls did not show activation with any of the peptides. CONCLUSION Patients with GSTT1 mismatch can develop a specific T-cell response, but the potential role of this response in the pathogenesis of plasma cell-rich rejection is unknown.

Genetic dissection of glutathione S-transferase omega-1:identification of novel downstream targets and Alzheimer's disease pathways

Alzheimer's disease(AD)is affected by genetic factors.Polymorphisms in the glutathione S-transfe rase omega-1(Gsto1)gene have been shown by genetic correlation analyses performed in different ethnic populations to be genetic risk factors for AD.Gene expression profile data from BXD recombinant inbred mice were used in combination with genetic and bioinformatic analyses to chara cterize the mechanisms underlying regulation of Gstol variation regulation and to identify network membe rs that may contribute to AD risk or progression.Allele-specific assays confirmed that variation in Gstol expression is controlled by cis-expression quantitative trait loci.We found that Gstol mRNA levels were related to several central nervous system traits,such as glial acidic fibrillary protein levels in the caudate putamen,co rtical gray matter volume,and hippocampus mossy fiber pathway volume.We identified 2168 genes whose expression was highly correlated with that of Gsto1.Some genes were enriched for the most common neurodegenerative diseases.Some Gsto1-related genes identified in this study had previously been identified as susceptibility genes for AD,such as APP,Grin2 b,Ide,and Psenen.To evaluate the relationships between Gstol and candidate network members,we transfected astrocytes with Gstol siRNA and assessed the effect on putative downstream effecto rs.We confirmed that knockdown of Gstol had a significant influence on Pa2g4 expression,suggesting that Pa2g4 may be a downstream effector of Gstol,and that both genes intera ct with other genes in a network during AD pathogenesis.

Association among Serum Organochlorine Pesticide Residues, Glutathione S-Transferase M1 Genetic Polymorphism and Female Breast Cancer

Background: The purpose of this study was to evaluate the association among serum organochlorine pesticide residues, glutathione S-transferase M1 genetic polymorphism and female breast cancer. Methods: A 1:1 matched case-control study of 140 newly diagnosed breast cancer patients and 140 non-cancer female patients who consulted the five largest hospitals in the Tangshan city from September 2006 to October 2007. Results: The result showed higher risk of breast cancer among subjects with higher levels of serum DDT and HCH residue, the OR was 3.18 (95%CI, 1.11 - 9.07) and 5.02 (95%CI, 1.64 - 16.56).The value of ORe associated with single environmental factor DDT high residues, and ORg associated with single GSTM1 deletion genotype were respectively 3.86 (1.20 - 12.47) and 1.34 (0.36 - 5.08). The OReg associated with combined action of two factors was 5.59 (1.63 - 18.90), and the value of interaction parameters (γ) equaled 1.24. The value of ORe associated with single environmental factor HCH higher residue and ORg associated with single GSTM1 deletion genotype were respectively 2.73 (0.84 - 8.87) and 1.48 (0.49 - 4.60). The value of OReg associated with combined action of two factors was 3.87 (1.18 - 12.68), and γ equaled 1.38. Conclusion: The results indicated that breast cancer occurrence was the combined result of environmental and genetic factors. The concurrent action of GSTM1 deletion genotype and DDT/HCH enhanced the risk of breast cancer.

Frequency of Null Phenotypes of Glutathione S-Transferase M1 and T1 among the Populations of Tabuk (Northwestern Part of Saudi Arabia)

Background: The variability in the distribution of the null phenotypes of GSTM1 and GSTT1, due to total or partial gene deletion resulting in the lack of the active enzyme, has been reported in different populations, especially in ethnically well-defined groups but not in Tabuk. This study investigated the variability in the distribution of the null phenotypes of GSTM1 and GSTT1 in the population of Tabuk (northwestern part of Saudi Arabia). Method: This study was conducted on 200 subjects of Tabuk—northwestern part of Saudi Arabia among which 100 were chronic smokers and 100 were nonsmokers. The subjects were reporting to hospital for routine checkup. All were without past history of any chronic disease and no significant abnormality. GST genotyping was done by multiplex PCR-based methods. The smoker and control groups were compared using a chi-square test with P GSTM1 deletion homozygosity of 14% and 1% was reported among non smokers and smokers, respectively whereas GSTT1 deletion homozygosity of 28% and 6% was reported among non smokers and smokers, respectively. Our results indicate that there are major differences in allelic distribution of GSTM1 and GSTT1 genes between the two groups investigated. Combined analysis of both genes revealed that 15% of smokers and non smokers harbor the deleted genotype of GSTM1 and 34% of smokers and non smokers harbor the deleted genotype of GSTT1 with significant differences. Conclusion: This study enables selecting subgroups among the general population who are more susceptible to DNA damage and will help genetic studies on the association of GST polymorphisms with disease risks and drug effects in Arab population. Studies with a larger sample size are needed to evaluate and confirm the validity of our results.

Association between Polymorphisms of Glutathione S-Transferase and Progression to Cervical Cancer in Women from Burkina Faso and Mali

Although persistence of high-risk human papillomavirus infection is the main risk factor, Glutathione S-Transferase highly polymorphic enzyme involved in the metabolism of xenobiotics, is a good candidate gene. The objective of this study was to compare the polymorphisms of Glutathione S-Transferase M1-null in women with cancerous lesions and without lesions. This study consisted of 322 uterine cervix samples of women from Mali and Burkina Faso with Cervical Intra-epithelial Neoplasia 2 and 3, adenocarcinoma and squamous cell carcinoma and 100 women with no lesions. Human Papillomavirus genotyping was performed by Real-time multiplex Polymerase Chain Reaction. Glutathione S-Transferase gene polymorphisms were determined using conventional Polymerase Chain Reaction followed by migration on agarose gel. A statistically significant association with high relative risks of 10.77 for the development of High grade Superficial or Squamous Intra-epithelial Lesion (95% CI = 5.59 - 20.72;p < 0.001), and 13.20 for cancer development (95% CI = 6.79 - 25.63;p < 0.001) was found in women with the null genotype of Glutathione S-Transferase M1 in the study population. In Burkina Faso and Mali, Glutathione S-Transferase M1-null presented relative risks of 9 and 11.05 for high-grade lesions, 15 and 11.40 for cancer. Similarly, significant results had been observed in women with human papillomavirus positive and human papillomavirus negative. The results of the present study support the idea that the deletion of Glutathione S-Transferase M1 plays a crucial role in the progression of high-grade lesions and cervical cancer.

Glutathione S-transferase P1 IlelO5Val Polymorphism and Male Infertility Risk： An Updated Meta-analysis

Background： Several studies concerning the association between glutathione S-transferase P1 （GSTP1） Ilel05Val polymorphism and male infertility risk have reported controversial findings. The present study was aimed to explore this association using a recta-analysis. Methods： The PubMed, EMBASE, China National Knowledge Infrastructure （CNKI）, and Wanfang databases were searched. Odds ratios （ORs） with 95% confidence intervals （C/s） were calculated to estimate the strength of the association. Results： A total of 3282 cases and 3268 controls in nine case-control studies were included. There was no significant association between GSTP1 llel05Val polymorphism and male infertility in the overall population, but significant associations were lbund under the dominant （OR = 1.23, 95% CI = 1.04-1.46, F = 32.2%） and heterozygote （OR = 1.29, 95% C1 - 1.08-1.53, F = 26.8%） models after excluding studies for which the data did not satisfy Hardy-Weinberg equilibrium （HWE）. Similarly, subgroup analyses revealed no significant association in Asians or Chinese population although a significant association was apparent among Chinese population in studies with HWE under the heterozygote model （OR = 1.25, 95% CI = 1.03-1.52, F = 44.1%）. Significant heterogeneity could be observed in some genetic models, but this heterogeneity was not significant when stratified by HWE. No evidence for publication bias was found. Conclusions： The GS-FP1 lle105Val polymorphism might not be associated with male infertility risk, and thus additional well-designed studies with larger sample size are warranted.

P-pg、MRP1和GST-π在宫颈鳞癌中的表达及对新辅助化疗敏感性的预测研究

目的探讨宫颈癌新辅助化疗前后P-gp、MRP1和GST-π的表达及其与化疗疗效的关系。方法运用S-P法检测30宫颈鳞癌新辅助化疗前后P-gp、MRP1和GST-π的表达水平。结果①化疗后宫颈鳞癌组织中P-gp阳性表达率为83.3%,显著高于化疗前55.5%(P<0.05);化疗前P-gp阴性患者有效率为100%显著高于P-gp表达阳性患者有效率64.1%(P<0.05)。②化疗后宫颈鳞癌组织中GST-π阳性表达率为83.3%,显著高于化疗前60%(P<0.05);化疗前P-gp阴性患者有效率为83.3%显著高于P-gp表达阳性患者有效率66.7%(P<0.05)。③化疗前后宫颈鳞癌组织中MRP1阳性表达率分别为为86.7%和93.3%,二者间无显著性差异(P>0.05);化疗前P-gp阴性、阳性患者有效率分别为100%和76.9%,二者间无显著性差异(P>0.05)。结论P-pg和GST-π可作为预测宫颈鳞癌化疗敏感性指标。

GSTP1、MTHFR基因多态性与PF方案治疗食管癌疗效相关性研究

目的：了解食管癌患者谷胱甘肽S转移酶P1（glutathione S-transferase Pi-1,GSTP1）,亚甲基四氢叶酸还原酶（methylenetetrahydrofolate reductase,MTHFR）基因多态性分布频率,探讨GSTP1、MTHFR基因多态性与食管癌患者PF方案（顺铂联合5-氟尿嘧啶）化疗疗效相关性。方法：从168例接受PF方案化疗的食管癌病例中采外周血提取DNA,采用PCR法扩增目的基因片段,直接测序法分析GSTP1、MTHFR基因多态性,每两周期化疗后评价疗效一次,按照实体瘤的疗效评价标准进行评价。结果：168例食管癌患者PF方案化疗疗效评价（实体瘤的疗效评价标准RECIST 3.0）,CR 19例（11.3%）,PR 60例（35.7%）,SD 48例（28.6%）,PD 41例（24.4）,食管癌患者GSTP1基因野生型（AA）有效率为22.0%,杂合型（AG）＋纯合型（GG）有效率为25.0%,MTHFR C667T基因野生型（CC）有效率为14.3%,杂合型（CT）＋纯合型（TT）有效率为32.7%,MTHFR A1298C基因野生型（AA）有效率为17.3%,杂合型（AC）＋纯合型（CC）有效率为29.8%,其中GSTP1基因野生型（AA）与杂合型（AG）＋纯合型（GG）的OR=2.520,95%CI：11.237-25.191,P=0.012,MTHFR C6 6 7 T基因野生型（CC）与杂合型（CT）＋纯合型（TT）的OR=1.933,95%CI：5.987-16.354,P=0.032,MTHFR A1298C基因野生型（AA）与杂合型（AC）＋纯合型（CC）的OR=3.514,95%CI：19.018-27.332,P=0.511。结果显示食管癌患者GSTP1基因及MTHFR C667T基因多态性与PF方案化疗后疗效相关,且杂合型及纯合型疗效优于野生型。结论：GSTP1及MTHFR-C677T基因多态性与5-氟尿嘧啶的疗效具有相关性,而MTHFR-A1298C基因多态性与5-氟尿嘧啶的疗效无明显相关性,故检测GSTP1及MTHFR-C677T基因多态性可预测PF方案化疗疗效。

FGFR1在乳腺癌分子亚型中的表达及意义

目的探讨成纤维细胞生长因子受体1(fibroblast growth factor receptor 1,FGFR1)在乳腺癌组织及正常组织中的表达及与雌激素受体(ER)、孕激素受体(PR)、人表皮生长因子受体2(HER2)、谷胱甘肽-S转移酶-π(GST-π)、拓扑异构酶Ⅱα(TOPOⅡα)间的关系。方法采用免疫组织化学S-P法检测88例乳腺浸润性导管癌和30例正常乳腺组织中FGFR1的表达情况。结果 FGFR1在浸润性导管癌及正常乳腺组织中的表达率分别为37.5%、10.0%,两者间差异有统计学意义(P<0.05);FGFR1在GST-π阳性及阴性乳腺浸润性癌患者中表达率分别为60.4%、10.0%,差异有统计学意义(P<0.05)。FGFR1在TOPOⅡα阳性及阴性乳腺癌患者中表达率分别为47.2%、22.9%,差异有统计学意义(P<0.05)。FGFR1在Luminal A型、Luminal B型、HER2过表达型、Basal-like型中表达率分别为25.0%、35.3%、56.3%、63.6%,FGFR1表达率在Luminal A和HER2过表达型、Luminal A和Basal-like型间差异有统计学意义(P<0.05)外,其他各组表达率间差异均无统计学意义(P>0.05)。FGFR1与ER、PR、HER2状态无相关性(P>0.05),但FGFR1与内分泌治疗耐药呈正相关(P<0.05)。结论 FGFR1在浸润性导管癌的不同分子分型中表达有差异,可能成为预测乳腺癌预后的重要指标之一。

Impact of SNP-SNP interactions of DNA repair gene ERCC5 and metabolic gene GSTP1 on gastric cancer/atrophic gastritis risk in a Chinese population

AIM To investigate the interactions of the DNA repair gene excision repair cross complementing group 5(ERCC5) and the metabolic gene glutathione S-transferase pi 1(GSTP1) and their effects on atrophic gastritis(AG) and gastric cancer(GC) risk.METHODS Seven ERCC5 single nucleotide polymorphisms(SNPs)(rs1047768, rs2094258, rs2228959, rs4150291, rs4150383, rs751402, and rs873601) and GSTP1 SNP rs1695 were detected using the Sequenom MassA RRAY platform in 450 GC patients, 634 AG cases, and 621 healthy control subjects in a Chinese population.RESULTS Two pairwise combinations(ERCC5 rs2094258 and rs873601 with GSTP1 rs1695) influenced AG risk(P_(interaction) = 0.008 and 0.043, respectively), and the ERCC5 rs2094258-GSTP1 rs1695 SNP pair demonstrated an antagonistic effect, while ERCC5 rs873601-GSTP1 rs1695 showed a synergistic effect on AG risk OR = 0.51 and 1.79, respectively). No pairwise combinations were observed in relation to GC risk. There were no cumulative effects among the pairwise interactions(ERCC5 rs2094258 and rs873601 with GSTP1 rs1695) on AG susceptibility(P_(trend) > 0.05). When the modification effect of Helicobacter pylori(H. pylori) infection was evaluated, the cumulative effect of one of the aforementioned pairwise interactions(ERCC5 rs873601-GSTP1 rs1695) was associated with an increased AG risk in the case of negative H. pylori status(P_(trend)= 0.043).CONCLUSION There is a multifarious interaction between the DNA repair gene ERCC5 SNPs(rs2094258 and rs873601) and the metabolic gene GSTP1 rs1695, which may form the basis for various inter-individual susceptibilities to AG.

Role of microRNA-21 in uveal melanoma cell invasion and metastasis by regulating p53 and its downstream protein

AIM： To reveal the insight mechanism of liver metastasis in uveal melanoma, we investigated cell functions of microRNA-21 in three different uveal melanoma cell lines and analyze the relationship of target gene p53 and its downstream targets which been found significant expression in our previous study.METHODS： Quantitative real-time polymerase chain reaction （qRT-PCR） was used to detect microRNA-21 expression in normal uveal tissue and uveal melanoma cell lines. Lenti-virus expression system was used to construct OCM-1, MuM-2B and M619 cell line with stable overexpression and inhibition of microRNA-21. In vitro cell function tests such as cell proliferation, cell apoptosis, cell circle and abilities of migration and invasion were examined by MTT, BrdU assay, flow cytometry, transwell assay and Matrigel invasion assay respectively. The target gene was predicted by bioinformatics and confirmed by using a dual luciferase reporter assay. The expression of p53 and its suspected downstream targets LIM and SH3 protein 1 （LASP1） and Glutathione S Transferase pi （GST-Pi） were determined by qRT-PCR in mRNA level and western blotting analysis in protein level. Finally, the effect of microRNA-21 in a xenograft tumor model was assessed in four-week-old BALB/c nude mice. RESULTS： Compared to normal uveal melanoma, expressions of microRNA-21 were significantly higher in uveal melanoma cell lines. Overexpression of microRNA-21 promoted proliferation, migration, and invasion of OCM-1, M619 and MuM-2B cells, while inhibition of microRNA-21 reveal opposite effects. Wild type p53 was identified as a target gene of microRNA-21-3p, and proved by dual luciferase reporter assay. Up-regulated microRNA-21 inhibited the expression of wild type p53 gene, and the increased expression of LASP1 in mRNA level and protein level, while down-regulated microRNA-21 presented opposite way. However, GST-pi showed the potential pattern as expected, but relative mRNA level showed no statistically significant difference in OCM-1 cells. Furthermore, the mRNA expression of GST-pi was decreased in microRNA-21 overexpressing MuM-2B, and increased in M619 cells with inhibition of microRNA-21. In vivo, inhibition of microRNA-21 reduced tumor growth with statistically significant difference.CONCLUSION： These findings provide novel insight into molecular etiology of microRNA-21 in uveal melanoma cell lines, and suggest that microRNA-21 might be a potential candidate for the diagnosis and prognostic factor of human uveal melanoma in future.

哺乳动物GSTP1基因研究进展

谷胱甘肽硫转移酶P1(GSTP1)基因属于谷胱甘肽硫转移酶家族(GSTs)的P类,能够编码硫转移酶家族的酶。GSTP1分布在过氧化物酶体中,通过催化半胱氨酸残基的硫谷胱甘肽化改变某些靶蛋白的结构或功能特征,是参与异生物质代谢的一个重要调控基因。本文综述了GSTP1基因的定位、结构、表达、分子机制及GSTP1参与的代谢反应,为深入研究GSTP1基因提供参考。

Genetic analysison the association between polymorphisms of UGT1A1and GST and neonatal hyperbilirubinemia

Objective:To investigate whether polymorphisms of uridine diphosphoglucuronate-glucuronosyltransferase 1A1(UGT1A1)c.211G>A and glutathione S transferases(GST)gene(GSTT1and GSTM1)are associated with neonatal hyperbilirubinemia.Methods:A case-control study was performed.A multiplex polymerase chain reaction(PCR)was used to detect the GSTT1and GSTM1polymorphisms.Single nucleotide polymorphism of UGT1A1c.211G>A was identified by PCR combined with DNA sequencing.The effects and co-expression of UGT1A1c.211G>A,GSTT1and GSTM1gene polymorphisms on the development of neonatal hyperbilirubinemia were estimated.Results:A allele frequency of c.211G>A polymorphism of UGT1A1gene was 0.186in case group and 0.086in control group,respectively.A allele frequency of the polymorphism of UGT1A1gene in the case group was higher than that of the control group(χ2=5.968,P=0.022).The frequencies of GSTT1and GSTM1in the case group were similar to that of the control groups.Logistic regression analysis showed that UGT1A1c.211G>A variant and UGT1A1+GSTM1 mutation affected neonatal hyperbilirubinemia.Conclusion:UGT1A1c.211G>A gene polymorphism may be one risk factor involved in the development of neonatal hyperbilirubinemia,and GST gene polymorphism may not be associated with the development of neonatal hyperbilirubinemia in our study.The co-expression of UGT1A1c.211G>A and GSTM1polymorphisms may reduce the risk for neonatal hyperbilirubinemia development.

木犀草素对白血病K562/ADR细胞多药耐药的逆转作用及机制研究

目的 探究木犀草素(Lut)对慢性髓系白血病K562/ADR细胞多药耐药性的逆转作用及其机制。方法K562和K562/ADR细胞经不同浓度阿霉素(ADR)处理24 h后,采用CCK-8实验检测K562/ADR细胞耐药倍数。Lut单独或联合ADR作用K562/ADR细胞24 h后,采用CCK-8实验检测Lut的细胞毒性及对ADR的增敏作用。取对数生长期K562/ADR细胞分为0μmol/L Lut组、2μmol/L Lut组、4μmol/L Lut组,采用流式细胞术检测细胞内ADR蓄积量的变化;RT-PCR法和Western blot法分别检测核因子E2相关因子2(Nrf2)、多药耐药相关蛋白1(MRP1)、P-糖蛋白(P-gp)、谷胱甘肽-S-转移酶pi(GST-pi)mRNA和蛋白的表达;谷胱甘肽(GSH)试剂盒检测细胞内GSH的含量。结果 与K562细胞相比,K562/ADR细胞株对ADR具有明显的耐药性,耐药倍数为53.69倍。与0μmol/L Lut相比,不同浓度Lut作用于K562/ADR细胞后,细胞生长均受到不同程度的抑制(P<0.05),其中2、4μmol/L Lut对K562/ADR细胞的增殖抑制率<10%,为无毒性的Lut浓度。与0μmol/L Lut组相比,2、4μmol/L Lut组可明显增强ADR对K562/ADR的细胞增殖抑制率,增加细胞内ADR蓄积量,提高逆转耐药倍数,降低细胞内GSH含量,下调细胞中MRP1、P-gp、GST-pi、Nrf2 mRNA及蛋白的表达(P<0.05);且4μmol/L Lut作用效果较2μmol/L Lut更显著。结论Lut可抑制K562/ADR细胞增殖,逆转其对ADR的耐药性,其机制可能与Lut下调Nrf2、MRP1、P-gp、GST-pi表达,进而增加细胞内ADR蓄积量有关。