Role of glutathione transferase in nonalcoholic fatty liver disease: An analysis based on gene expression profile / 临床肝胆病杂志

Objective To investigate the role of glutathione transferase in nonalcoholic fatty liver disease (NAFLD) induced by high-fat diet using the RNA-Seq technique in combination with gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of differentially expressed genes. Methods A total of 14 male C57BL/6J mice were divided into control group with 6 mice and model group with 8 mice by random sampling. The mice in the control group were fed with normal diet, and those in the model group were fed with high-fat diet for 7 consecutive weeks to establish a model of NAFLD. Kits were used to measure the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the level of triglyceride (TG), and HE staining and oil red staining were used to observe liver pathology and deposition of lipid droplets. Liver tissue RNA was extracted for RNA-Seq, and genes with a fold change of ≥2.0 and a P value of 0.05). Compared with the control group, the model group had a significantly higher serum level of TG (2.02±0.50 mmol/L vs 1.00±0.29 mmol/L, t =-4.45, P =0.001). HE staining showed diffuse steatosis and ballooning degeneration in the model group, and oil red staining showed that the model group had a significant increase in orange-red lipid droplets in the cytoplasm of hepatocytes and a significantly higher grade of hepatocyte steatosis than the control group (1.88±0.64 vs 1.00±0.00, t =-3.86, P =0.006). RNA-seq results showed a total of 1367 differentially expressed genes between the two groups, among which there were 608 upregulated genes and 759 downregulated genes, and there were 17 differentially expressed GST genes between the two groups. The top 10 GST genes in terms of fold change were validated, and compared with the control group, the model group had downregulated expression of GSTa2, GSTa3, GSTa4, GSTm1, GSTm2, GSTm3, GSTm4, GSTp1, and GSTo1 and upregulated expression of GSTk1. The results of qRT-PCR were consistent with the results of sequencing. Conclusion GST affects lipid metabolism by participating in various biological processes such as steroid metabolism, fatty acid metabolism, and cholesterol metabolism and is closely associated with the pathogenesis of NAFLD.

The strategy change of targeting iron homeostasis for leukemia therapy: from iron chelation to iron overload as a therapeutic strategy to induce ferroptosis / 白血病·淋巴瘤

Iron, an indispensable element for life, is involved in all kinds of vital physiological activities. Due to its potential toxicity, the body has a strict regulation mechanism of iron metabolism to maintain the "iron homeostasis". Dysregulation of iron metabolism and subsequent accumulation of excess iron are closely associated with the development and progression of leukemia. Specifically, due to the pro-oxidative nature of iron and its damaging effects on DNA, excess iron promotes the progression of leukemia; on the other hand, leukemia cells need to obtain more iron than normal cells to maintain rapid growth and proliferation, which is known as "iron addiction". Iron chelators can remove iron in leukemia cells and induce differentiation and apoptosis of leukemia cells. However, "iron addiction" makes leukemia cells more susceptible to iron overload, and is more sensitive to a new form of iron-catalyzed cell death which was named ferroptosis. According to the different needs of leukemia cells and normal cells for iron, the method of selectively killing leukemia cells through iron overload may become a new strategy for leukemia treatment. This paper reviews the strategy of targeting iron homeostasis for leukemia therapy.

Análise comparativa do polimorfismo genético da glutationa transferase, do Helicobacter pylori e do vírus Epstein-Barr entre a área do tumor e as margens de ressecção proximal e distal do câncer gástrico / Comparative analysis of glutathione transferase genetic polymorphism, Helicobacter pylori and Epstein-Barr virus between the tumor area and the proximal and distal resection margins of gastric cancer

RESUMO Objetivo: comparar o polimorfismo dos genes Glutationa S-transferase teta 1 (GSTT1) e Glutationa S-transferase mu 1 (GSTM1) da área do tumor com as margens proximal e distal de espécimes de estômago ressecados de pacientes com câncer gástrico, e investigar a presença do DNA do vírus Epstein-Barr (EBV) e Helicobacter pylori. Métodos: coletamos prospectivamente amostras teciduais da área do tumor e das margens de ressecção proximal e distal dos estômagos de dez pacientes com adenocarcinoma gástrico submetidos à gastrectomia com linfadenectomia D2 e submetemos esses espécimes à extração de DNA. Comparamos a área do tumor com as margens proximal e distal dos estômagos ressecados para o polimorfismo dos genes GSTT1 e GSTM1 e investigamos a presença de DNA do EBV e H. pylori. Utilizamos o exon 5 do gene p53 como controle interno da reação de PCR multiplex. Resultados: em um paciente, detectamos genótipos GSTT1 e GSTM1 nulos na área do tumor, em contraste com a presença de ambos os genes nas margens proximal e distal. Encontramos DNA do EBV e H. pylori na área do tumor e também nas margens proximal e distal. Em outro paciente, a margem proximal foi negativa para GSTT1 e o DNA do EBV foi negativo na margem distal. Em três pacientes, o EBV-DNA foi negativo apenas na margem distal. Conclusão: este é o primeiro relato em que diferentes genótipos, infecção por EBV-DNA e H. pylori foram observados no mesmo paciente, indicando provável deleção desses genes em resposta à progressão tumoral e heterogeneidade intratumoral.

ABSTRACT Objective: to compare the polymorphism of the Glutathione S-transferase theta 1 (GSTT1) and Glutathione S-transferase mu 1 (GSTM1) genes from the tumor area with the proximal and distal margins of stomach specimens resected from patients with gastric cancer, and to investigate the presence of Epstein-Barr virus (EBV) DNA and Helicobacter pylori. Methods: we prospectively collected tissue specimens from the tumor area and from the proximal and distal resection margins of the stomachs of ten patients with gastric adenocarcinoma who underwent gastrectomy with D2 lymphadenectomy, and submitted these specimens to DNA extraction. We compared the tumor area with the proximal and distal margins of the resected stomachs for polymorphism of GSTT1 and GSTM1 genes and investigated the presence of EBV-DNA and H. pylori. We used the p53 exon 5 gene as an internal control of the multiplex PCR reaction. Results: in one patient, we detected null GSTT1 and GSTM1 genotypes in the tumor area, in contrast to the presence of both genes in the proximal and distal margins. We found EBV-DNA and H. pylori in the tumor area and also in the proximal and distal margins. In another patient, the proximal margin was negative for GSTT1, and EBV-DNA was negative in the distal margin. In three patients, EBV-DNA was negative only in the distal margin. Conclusion: this is the first report where different genotypes, EBV-DNA and H. pylori infection were observed in the same patient, indicating a probable deletion of these genes in response to tumor progression and intratumoral heterogeneity.

Association between the glutathione S-transferase P1 (GSTP1) Ile105Val gene polymorphism in obese and overweight patients over 60 years / Associação entre o polimorfismo Ile105Val do gene da glutationa S-transferase P1 (GSTP1) em pacientes obesos e com sobrepeso acima de 60 anos

ABSTRACT INTRODUCTION: Obesity is related to the possibility of a number of metabolic damage associated with oxidative stress. The enzymes of the glutathione S-transferase (GST) family have the function of promoting detoxification; however, polymorphisms in the glutathione S-transferase P1 (GSTP1) gene generate less efficient alleles as well as a decrease in their amount and activity. OBJECTIVE: This study aimed to analyze the frequency of the alleles (A and G) and the genotypes of the GSTP1 Ile105Val gene polymorphism, and its association with obesity in the elderly. MATERIALS AND METHODS: This was a cross-sectional study involving 232 subjects aged between 60-98 years, of both sexes, originating from southern Brazil. The volunteers were categorized according to the body mass index (BMI) in three groups: normal weight (n = 52), overweight (n = 133), and obese (n = 47). Anthropometry was evaluated and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used for genetic analysis, from peripheral blood samples. RESULTS: The allelic frequency in the elderly obese group was 37.2% for A and 62.8% for G allele, and the genotypic frequency observed was AA 8.5%, AG 57.4% and GG 34.1%. Both the G allele as the GG and AG genotypes were significantly higher in the obese group compared to the other groups (p < 0.001). CONCLUSION: A higher prevalence of the G allele was observed in elderly obese group, responsible for encoding an abnormal enzyme and consequent reduction of antioxidant defenses, which contribute to inflammation process and obesity in the elderly.

RESUMO INTRODUÇÃO: A obesidade está relacionada com a possibilidade de numerosos danos metabólicos associados ao estresse oxidativo. As enzimas da família glutationa S-transferase (GST) têm como função promover a detoxificação, entretanto, polimorfismos no gene da glutationa S-transferase P1 (GSTP1) geram alelos menos eficientes, bem como diminuição da sua quantidade e atividade. OBJETIVO: Este estudo teve como objetivo analisar a frequência dos alelos (A e G) e dos genótipos do polimorfismo Ile105Val do gene GSTP1, além de sua associação à obesidade em idosos. MATERIAIS E MÉTODOS: Tratou-se de um estudo transversal, o qual envolveu 232 indivíduos com idades entre 60 e 98 anos, de ambos os sexos, oriundos da região Sul do Brasil. Os voluntários foram caracterizados de acordo com o índice de massa corporal (IMC) em três grupos: peso normal (n = 52), sobrepreso (n = 133) e obesos (n = 47). A antropometria foi avaliada, e a técnica de reação em cadeia da polimerase-polimorfismo no comprimento de fragmentos de restrição (PCR-RFLP) foi usada para análise genética a partir de amostras de sangue periférico. RESULTADOS: A frequência alélica no grupo de idosos obesos foi de 37,2% para o alelo A e 62,8% para o G, e a frequência genotípica observada, de AA 8,5%, AG 57,4% e GG 34,1%. Tanto o alelo G quanto os genótipos GG e AG foram significativamente maiores no grupo obeso quando comparados com os dos demais grupos (p < 0,001). CONCLUSÃO: Observou-se maior prevalência do alelo G no grupo de idosos obesos, responsável pela codificação de uma enzima anormal e consequente diminuição das defesas antioxidantes, que contribuem para o processo inflamatório e a obesidade em idosos.

Effect of TRAIL on Expression of Multidrug Resistance Gene GST-π in Drug-resistant Human Gastric Cancer Cell Line SGC7901/VCR / 胃肠病学

Background:Multidrug resistance of tumor cells is one of the important factors that cause failure of chemotherapy in advanced gastric cancer. Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)may enhance the killing effect of chemotherapeutics on tumor cells,and reverse drug-resistant cell lines to sensitive cell lines,but its mechanism is not yet clear. Aims:To study the effect of TRAIL on expression of multidrug resistance gene glutathione S-transferase-π(GST-π)in drug-resistant human gastric cancer cell line SGC-7901 / VCR and the potential mechanism of TRAIL in reversing multidrug resistance of gastric cancer cells. Methods:SGC-7901 / VCR cells were treated with TRAIL in different doses (50,100,200 and 400 μg/ L)for 48 hours. After treatment,expression of GST-π mRNA in SGC-7901 / VCR cells and concentration of GST-π in culture supernatant were detected by RT-PCR and ELISA,respectively. Results:TRAIL could inhibit mRNA expression and protein secretion of GST-π in SGC-7901 / VCR cells in a dose-dependent manner within a certain range(≤200 μg/ L). The relative expression levels of GST-π mRNA in 50,100,200 and 400 μg/ L TRAIL groups were 0. 89 ± 0. 04,0. 77 ± 0. 08,0. 65 ± 0. 06 and 0. 61 ± 0. 03,respectively,and the concentrations of GST-π in culture supernatant in these groups were(57. 56 ± 1. 19)ng/ mL,(56. 30 ± 0. 80)ng/ mL,(31. 41 ± 1. 65)ng/ mL and (30. 80 ± 1. 34)ng/ mL,respectively,all were significantly lower than those in control group[1. 01 ± 0. 13 and(58. 62 ± 1. 38)ng/ mL,P all < 0. 05]. Conclusions:TRAIL may play a potential role in reversing multidrug resistance of gastric cancer cells through down-regulating GST-π expression.

The correlations of LRP and GST-π to chemotherapy resistance and prognosis of ovarian cancer / 中国医师杂志

Objective To explore the correlations of lung resistance protein (LRP) and glutathione S transferase π (GST-π) to chemotherapy resistance and prognosis of epithelial ovarian cancer.Methods The expressions of LRP and GST-π in epithelial ovarian cancer were examined with immunohistochemistry.Correlations of LRP and GST-π to chemotherapy efficacy and survival time after operation were analyzed.Results The short-term efficacy rates of ovarian cancer were lower in patients with positive expressions of LRP and GST-π than those with negative expressions [61.2％,61.7％ vs 94.1％,89.5％,x2 =6.47,4.94,P =0.011,P =0.026].The positive rates of LRP and GST-π were significant higher in patients with chemotherapy resistance than in those sensitive to chemotherapy [91.3％,87.0％ vs 65.1％,62.8％,P ＜ 0.05].Log-rank test showed that patients with positive LRP and GST-π had shorter survival time than those negative,and patients with both positive LRP and GST-π had shorter survival time than those both negative (P ＜ 0.05).Conclusions The expressions of LRP and GST-π in epithelial ovarian cancer could be used to predict chemotherapy resistance and prognosis of patients.

Correlation between Polymorphisms of GSTM1, GSTT1 and GSTP1 (rs1695) on Hematologic Toxicities with Anthracycline/Paclitaxel-Based Chemotherapy in Breast Cancer / 天津医药

Objective To examine the effects of genetic polymorphisms in GSTM1, GSTT1 and GSTP1 (rs1695) genes on hematologic toxicities of breast cancer patients receiving Anthracycline/Paclitaxel- based chemotherapy. Methods Multiply PCR technique and High Resolution Melting method were used to examine these 3 genes polymorphsim in female breast cancers (n=252). Results The GSTP1(rs1695) AG/GG genotype was a risk factor forⅢ-Ⅳdegree of neu-trophil toxicity when patients received Paclitaxel-based chemotherapy and Anthracycline-Paclitaxel-based chemotherapy (OR=6.111, 95%CI 1.526-24.469, P0.05);There was no statistic difference onⅢ-Ⅳdegree hematologic toxicities rates between GSTM1(+) and GSTM1(-), GSTT1(+) and GSTT1(-), GSTP1AA and GSTP1AG/GG patients after they receiced Anthracycline-based chemotherapy (P>0.05). Conclusion The genetic polymorphisms in GSTP1(rs1695) can be used as a gene marker for forecasting the chemotherapy, inducing neutrope-nia toxicities in breast cancer patients receiving Paclitaxel-based chemotherapy.

Value of serum GSTP1 gene quantitative methylation analysis for early diagnosis of hepatocellular carcinoma / 国际检验医学杂志

Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .

Polymorphisms of GSTM1 and GSTT1 genes in breast cancer susceptibility: a case-control study / Polimorfismo dos genes GSTM1 e GSTT1 na suscetibilidade do câncer de mama: estudo caso-controle

PURPOSE: This study aimed to evaluate the frequency of homozygous deletion of GSTM1 and GSTT1 genes and their combinations between patients with breast cancer and healthy individuals, associating them with disease susceptibility. METHODS: This is a case-control study in which 49 women diagnosed with breast cancer confirmed by pathological examination and 49 healthy women with no evidence of cancer and no prior family history of breast cancer were invited to participate. All of them answered a questionnaire with epidemiological data and were submitted to blood sample collection. Genomic DNA was extracted from blood, and genotyping was performed by polymerase chain reaction. Data were analyzed with SPSS 20.0. RESULTS: The frequency of null alleles for GSTM1 and GSTT1 was 58.8 and 61.7%, respectively, for patients with breast cancer, and 41.2 and 38.3%, respectively, in control patients. In homozygous deletion of the GSTM1 gene, a significantly higher frequency was found in the breast cancer cases. CONCLUSION: Breast cancer patients presented higher frequency of homozygous deletion of the GSTM1 gene compared with the control group.

OBJETIVO: Este estudo teve como objetivo avaliar a frequência de deleção homozigótica dos genes GSTM1 e GSTT1 e suas combinações entre os pacientes com câncer de mama e indivíduos saudáveis, associando-se a suscetibilidade à doença. MÉTODOS: Este é um estudo de caso-controle, no qual 49 mulheres diagnosticadas com câncer de mama confirmado por exame anatomopatológico e 49 mulheres saudáveis, sem evidência de câncer e sem história familiar prévia de câncer de mama, foram convidadas a participar. Todas responderam a um questionário com dados epidemiológicos e foram submetidas à coleta de sangue. O DNA foi extraído a partir de sangue, e genotipagem foi realizada por reação em cadeia da polimerase. Os dados foram analisados com o SPSS 20.0. RESULTADOS: A frequência de alelos nulos para GSTM1 e GSTT1 foi de 58,8 e 61,7%, respectivamente, para as pacientes com câncer de mama, e 41,2 e 38,3%, respectivamente, em pacientes do grupo controle. Em deleção homozigótica do gene GSTM1, uma frequência significativamente maior foi encontrada nos casos. CONCLUSÃO: Pacientes com câncer de mama apresentam uma maior frequência de deleção homozigótica do gene GSTM1 quando comparadas com o grupo controle.

Nullity of GSTT1/GSTM1 related to pesticides is associated with Parkinson's disease / Nulidade de GSTT1/GSTM1 relacionada a pesticidas associa-se com doenca de Parkinson

Genetic and environmental factors affect the pathogenesis of Parkinson's disease (PD). Genetic variants of the enzyme glutathione S-transferases (GST) may be related to the disease. This study aimed to evaluate the influence of genetic variants of GST (GSTT1/GSTM1) and their association with the exposure to environmental toxins in PD patients. We studied 254 patients with PD and 169 controls. The GSTM1/GSTT1 variants were analyzed by polymerase chain reaction. We applied the Fisher's exact test and the χ2 test for statistical analysis (p<0.05). The present and absence for GSTT1 and GSTM1 were similar in patients and controls. The null for GSTT1 and GSTM1 (0/0) and exposure to pesticides prevailed in patients (18%) compared to controls (13%, p=0.014). This study suggests the association between PD and previous exposure to pesticides, whose effect may be enhanced in combination with null for GSTT1/GSTM1.

Fatores genéticos e ambientais influenciam a patogênese da doença de Parkinson (DP). Variantes genéticas das enzimas glutationa S-transferases (GST) parecem estar envolvidas com a doença. Os objetivos deste estudo foram avaliar a influência de variantes genéticas de GST (GSTT1/GSTM1) e sua associação com exposição a toxinas ambientais em pacientes com DP. Foram estudados 254 pacientes com DP e 169 controles. As variantes para GSTM1/GSTT1 foram analisadas por reação em cadeia da polimerase. Para análise estatística foram aplicados os testes de Fisher e do χ2 (p<0,05). Tanto a presença quanto a nulidade para GSTT1 e GSTM1 foram semelhantes em pacientes e controles. A nulidade para GSTT1 e GSTM1 (0/0) e contato com agrotóxicos prevaleceu nos pacientes (18%) em relação aos controles (13%, p=0,014). Este estudo sugere associação entre DP e contato prévio com agrotóxicos, cujo efeito parece potencializado em combinação com nulidade para GSTT1/GSTM1.

Exposure to pesticides and heterozygote genotype of GSTP1-Alw26I are associated to Parkinson's disease / Exposição a pesticidas e genótipo heterozigoto de GSTP1-Alw26I associam-se à doença de Parkinson

Objective This study aimed to analyze the frequency of GSTP1-Alw26I polymorphism and to estimate its association with toxic substances in Parkinson's disease (PD). Methods A study group with 154 patients - subdivided into familial and sporadic PD groups - and 158 elderly individuals without the disease (control group) were evaluated. GSTP1-Alw26I polymorphism was analyzed by polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP). Results Patients were significantly more exposed to pesticides compared with the control group (p=0.0004), and the heterozygote genotype associated to exposure to pesticides also prevailed in patients (p=0.0001). Wild homozygote genotype was related to tobacco use (p=0.043) and alcoholism (p=0.033) in familial PD patients. Conclusion Exposure to pesticides is associated to PD, whose effect can be enhanced when combined with the heterozygote genotype of GSTP1-Alw26I. Also, large genetic and environmental studies considering tobacco use, alcoholism, GSTP1 and PD are necessary to confirm our findings. .

Objetivo Analisar a frequência do polimorfismo GSTP1-Alw26I, assim como estimar sua associação com substâncias tóxicas na doença de Parkinson (DP). Métodos A casuística avaliada foi composta por um grupo de estudo, com 154 pacientes, subdivididos em DP familial e esporádica, e outro com 158 idosos sem a doença (grupo controle). O polimorfismo GSTP1-Alw26I foi analisado por reação em cadeia da polimerase/polimorfismo de comprimento do fragmento de restrição (PCR/RFLP). Resultados Os pacientes foram significativamente mais expostos a pesticidas, comparados com o grupo controle (p=0,0004), e o genótipo heterozigoto associado a exposição a pesticidas também prevaleceu nos pacientes (p=0,0001). O genótipo homozigoto selvagem apresentou relação com tabagismo (p=0,043) e etilismo (p=0,033) em pacientes com DP familial. Desse modo, a exposição a pesticidas está associada à DP, cujo efeito pode ser potencializado quando combinado ao genótipo heterozigoto de GSTP1-Alw26I. Estudos genético-ambientais envolvendo tabagismo, etilismo, GSTP1 e DP devem ser realizados em casuísticas numerosas, confirmando essa associação. .

Study on the relationship between heme oxygenase-1 and glutathione S-transferase and cerebral atherosclerosis / 中国医师杂志

Objective To investigate the relationship between heme oxygenase-1 (HO-1),glutathione S-transferase (GST) and cerebral atherosclerosis.Methods Cerebralvascular status was assessed with color flow Doppler sonography,transcranial Doppler (TCD),magnetic resonance angiography (MRA)or/and digital subtraction angiography (DSA) in patients with cerebral atherosclerosis (mild,moderate,and severity).Serum HO-1 and GST were measured with enzyme-linked immunosorbent assay (ELISA).Results In comparison between case and control groups,there was significant difference in age,hypertension,cerebral infarction,uric acid,and HO-1 (P =0.041,0.008,0.000,0.036,and 0.001).The level of serum HO-1 in the severe atherosclerosis was lower than that in the mild and moderate atherosclerosis (P =0.000 and 0.002).Logistic regression was used to find the association of HO-1 and the degree of cerebral atherosclerosis (P =0.000).Conclusions HO-1 might be related to cerebral atherosclerosis.

Mutações do gene cystic fibrosis transmembrane conductance regulator e deleções dos genes glutationa S-transferase em pacientes com fibrose cística no Brasil / Cystic fibrosis transmembrane conductance regulator gene mutations and glutathione S-transferase null genotypes in cystic fibrosis patients in Brazil

OBJETIVO: Determinar os efeitos que a mutação do gene cystic fibrosis transmembrane conductance regulator (CFTR) e da deleção dos genes glutationa S-transferase (GST) mu-1 (GSTM1) e teta-1 (GSTT1) têm na evolução clínica da fibrose cística (FC) em pacientes da região sudeste do Brasil. MÉTODOS: Entre março de 2002 e março de 2005, incluímos no estudo todos os pacientes com FC atendidos consecutivamente no Departamento de Pediatria do Hospital de Clínicas da Faculdade de Ciências Médicas da Universidade Estadual de Campinas. O DNA genômico de 66 pacientes com FC foi analisado por PCR e digestão com endonuclease de restrição para a identificação dos genótipos. RESULTADOS: A mutação ΔF508 do gene CFTR foi identificada em 44 (66,7 por cento) pacientes. As deleções dos genes GSTM1, GSTT1 e da combinação nula GSTM1/GSTT1 foram identificadas em 40,9 por cento, 15,2 por cento e 3,0 por cento dos pacientes, respectivamente. A mutação ΔF508 do gene CFTR foi mais comum em pacientes diagnosticados com FC antes dos 2,5 anos de idade que naqueles diagnosticados mais tarde (75,5 por cento vs. 41,2 por cento; p = 0,008). CONCLUSÕES: Foram observadas frequências similares da mutação ΔF508 do gene CFTR e dos genótipos GSTM1 e GSTT1 nos pacientes, independentemente do sexo, etnia ou status da doença pulmonar ou pancreática. Quando os pacientes foram estratificados por aspectos clínicos e epidemiológicos, as frequências dos genótipos GSTM1 e GSTT1 nulos foram semelhantes, sugerindo que a ausência herdada dessas vias enzimáticas não altera o curso da FC. Em contraste, a alta frequência da mutação ΔF508 no gene CFTR encontrada em pacientes mais jovens sugere que essa mutação influencia a idade no momento do diagnóstico de FC nessa região do país.

OBJECTIVE: To determine the effects that mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene and deletion of the glutathione S-transferase (GST) genes mu-1 (GSTM1) and theta-1 (GSTT1) have on the clinical course of cystic fibrosis (CF) in patients residing in the southeastern region of Brazil. METHODS: The study sample consisted of all consecutive CF patients treated at the Hospital de Clínicas School of Medical Sciences of the State University at Campinas between March of 2002 and March of 2005. We included 66 CF patients. Genomic DNA was analyzed by polymerase chain reaction and restriction endonuclease digestion for the identification of the genotypes. RESULTS: The DF508 mutation of the CFTR gene was found in 44 patients (66.7 percent). The null genotypes GSTM1, GSTT1 and GSTM1/GSTT1 were found in 40.9 percent, 15.2 percent, and 3.0 percent of the patients, respectively. The DF508 CFTR mutation was more common in patients diagnosed with CF before 2.5 years of age than in those diagnosed later (75.5 percent vs. 41.2 percent; p = 0.008). The frequency of the DF508 CFTR mutation, as well as of the GSTM1 and GSTT1 genotypes, was not found to be associated with gender, ethnicity, pulmonary disease status, or pancreatic disease status. CONCLUSIONS: When the patients were stratified by clinical and epidemiological features, the frequencies of the GSTM1 and GSTT1 null genotypes were similar, suggesting that the inherited absence of these enzymatic pathways does not alter the course of CF. However, the high frequency of the DF508 CFTR mutation found in younger children suggests that it influences the age at diagnosis of CF in this region of Brazil.

Relationship of polymorphisms of AhR - 1661G/A with GSTP1 - 313A/G and susceptibility to endometriosis / 中华妇产科杂志

Objective To investigate the association of polymorphisms of arylhydrocarbon receptor (AhR) - 1661G/A with glutathione S-transferase pi ( GSTP1 ) - 313A/G and the susceptibility to endometriosis in southern Han Chinese.Methods Total of 432 endometriosis patients undergoing laparoscopic or laparotomy surgery matched with 493 patients with fallopian tube ligation,tubal recanalization,laparoscopic hydrotubation,benign ovarian tumor and teratoma surgeries without endometriosis as control group were enrolled in this study.The single nucleotide polymorphism (SNP) of AhR -1661G/A and GSTP1 -313A/G were detected by using a fluorescent quantitative PCR-based high resolution melting (HRM).Results The numbers of combined genotypes AhR - 1661G/A and GSTP1 -313A/G were 120 patients with AG + AA,64 patients with AG + AG,8 patients with AG + GG,109 patients with GG +AA,84 patients with GG + AG,4 patients with GG + GG,31 patients with AA + AA,10 patients with AA + AG,1 patient with AA + GG at endometriosis group and 131 patients with AG + AA,68 patients with AG + AG,6 patients with AG + GG,157 patients with GG + AA,66 patients with GG + AG,4 patients with GG + GG,35 patients with AA + AA,20 patients with AA + AG,3 patients with AA + GG at endometriosis group.There was no statistically different frequencies of genotypes between endometriosis group and control group (x2 = 12.558,P = 0.128 ).Compared with genotype GG + AA,the risk of endometriosis with genotype GG + AG was increased 1.833 time (95％CI:1.233-2.274).Conclusion The combined genotype GG + AG [ from AhR - 1661G/A (GG) and GSTP1 - 313A/G (AG) ] might be related with susceptibility to endometriosis.

Prevalence of glutathione S-transferase gene deletions and their effect on sickle cell patients

BACKGROUND: Glutathione S-transferase gene deletions are known detoxification agents and cause oxidative damage. Due to the different pathophysiology of anemia in thalassemia and sickle cell disease, there are significant differences in the pathophysiology of iron overload and iron-related complications in these disorders. OBJECTIVE: The aim of this study was to estimate the frequency of the GSTM1 and GSTT1 genotypes in sickle cell disease patients and their effect on iron status. METHODS: Forty sickle cell anemia and sixty sickle ß-thalassemia patients and 100 controls were evaluated to determine the frequency of GST gene deletions. Complete blood counts were performed by an automated cell analyzer. Hemoglobin F, hemoglobin A, hemoglobin A2 and hemoglobin S were measured and diagnosis of patients was achieved by high performance liquid chromatography with DNA extraction by the phenol-chloroform method. The GST null genotype was determined using multiplex polymerase chain reaction and serum ferritin was measured using an ELISA kit. Statistical analysis was by EpiInfo and GraphPad statistics software. RESULTS: An increased frequency of the GSTT1 null genotype (p-value = 0.05) was seen in the patients. The mean serum ferritin level was higher in patients with the GST genotypes than in controls; this was statistically significant for all genotypes except GSTM1, however the higher levels of serum ferritin were due to blood transfusions in patients. CONCLUSION: GST deletions do not play a direct role in iron overload of sickle cell patients.

Farmacogenética aplicada al tratamiento de la leucemia linfoide aguda / Pharmacogenetics applied to the treatment of acute lymphoid leukemia

Las enzimas de biotransformación y eliminación de los fármacos en pacientes con leucemia linfoide aguda tienen una acción determinante en el efecto terapéutico de los medicamentos antineoplßsicos. La presencia y actividad de estos complejos enzimáticos está codificada genéticamente y sujeta a variaciones alélicas, cuyas frecuencias son variables en las diferentes poblaciones humanas. Este polimorfismo genético influye sobre la efectividad terapéutica de los medicamentos y condiciona la carencia de toxicidad o presencia de esta, que en ocasiones puede ser fatal. Las enzimas tiopurin-metil-transferasa, metilén-tetrahidrofolato-reductasa y glutatión-tranferasa son sistemas destoxificadores de algunos de los quimioterápicos empleados en el tratamiento de la leucemia linfoide aguda. En este trabajo se revisan las características genéticas de estas enzimas, la frecuencia de sus polimorfismos y las implicaciones clínicas de su expresión. De igual modo se discute la importancia y los beneficios del genotipaje previo al inicio del tratamiento, con el fin de modificar las dosis de los medicamentos para optimizar su efecto terapéutico y disminuir su toxicidad. La farmacogenética constituye un área de creciente interés que ha tenido un desarrollo considerable en los últimos años, su conocimiento e implementación nos colocará en el camino de la medicina personalizada

The biotransformation and elimination enzymes of drugs in patients suffering from acute lymphoid leukemia play a decisive role on the therapeutical effect of anti-neoplastic drugs. The presence and activity of these enzymatic complexes are genetically coded and subjected to allele variations, the frequency of which is variable in the different human populations. This genetic polymorphism has an impact on the therapeutic effectiveness of drugs and determines the lack or the existence of toxicity that may sometimes become lethal. The enzymes called thiopurine-methyltransferase, methylen-tetrahydropholate-reductase and glutathione-transferase are detoxifying systems of some of the chemotherapeutic drugs that are used for the treatment of acute lymphoid leukemia. This paper reviewed the genetic characteristics of the enzymes, the frequency of polymorphisms and the clinical implications of their expression. Similarly, the importance and the benefits of genotyping before the treatment were discussed in order to change the drug doses to maximize the therapeutic effect and reduce toxicity. Pharmacogenetics has experienced great development in the last few years and draws growing interest; the knowledge about and the implementation of this discipline will take us to the customized medicine

Avaliação da glutationaS-transferaseM1 (GSTM1) e seu polimorfismo GSTM1 nulo na resposta ao traftamento com quimioterapia do câncer avançado de ovário / Assessment of glutathione S-transferaseM1 (GSTM1) and its polymorphisms GSTM1 null in the response to treatment with chemotherapy in advanced ovarian carcinoma

Objective: To assess if the genotype of the glutathione S-transferase M1 (GSTM1) enzyme and its GSTM1 null polymorphism can influence the response to chemotherapeutic treatment of advanced ovarian cancer. Methods: Case-control study of 112 patients with advanced ovarian cancer submitted to chemotherapy during the period from January 1995 to December 2005. The tissue to study the GSTM1 genotype and its deletion came from surgical staging to treat ovarian cancer. The PCR product generates two distinct genotypes, characterized as positive and null. The response to chemotherapy was evaluated using the World Health Organization (WHO) criteria.Patients were classified as having: a) no response, b) a response. Results: The presence of GSTM1 or its GSTM1 null polymorphism did not influence the preoperative chemotherapy response. Among the patients who did respond, 88.9% presented with positive GSTM1 and 11.1% with null GSTM1. Among the patients that did not respond, 85.71% presented with positive GSTM1 and 14.29% with null GSTM1 (p = 0.825). GSTM1 and its GSTM1 null polymorphism had no influence on the postoperative response to chemotherapy. Among the patients who did respond, 80.65% presented with positive GSTM1 and 19.35% with null GSTM1. Among the patients who did not respond, 87.50% presented with positive GSTM1 and 12.5% with the null polymorphism (p = 0.553). Conclusion: No difference was observed in the response to treatment with chemotherapy in patients with advanced ovarian cancer, as to the GSTM1 genotype compared to its GSTM1 null polymorphism.

Objetivo: Avaliar se o genótipo da enzima glutationa S-transferase M1 (GSTM1) e seu polimorfismo GSTM1 nulo podem influenciar na resposta ao tratamento quimioterápico do câncer avançado de ovário. Métodos: Estudo caso-controle de 112 pacientes portadoras de câncer avançado de ovário, submetidas a tratamento por quimioterapia no período de Janeiro de 1995 a Dezembro de 2005. O tecido para estudo do genótipo da GSTM1 e sua deleção foram provenientes do estadiamento cirúrgico para tratamento do câncer de ovário. O produto do PCR gera dois genótipos distintos, sendo caracterizado como positivo e nulo. A resposta à quimioterapia foi avaliada usando os critérios da Organização Mundial da Saúde. As pacientes foram classificadas em: a) sem resposta, b) com resposta. Resultados: A presença da GSTM1 ou seu polimorfismo GSTM1 nulo não influenciou na resposta à quimioterapia pré-operatória. Dentre as pacientes que responderam 88,9% apresentavam GSTM1 e 11,1% GSTM1 nulo. Dentre as pacientes que não responderam 85,71% apresentavam GSTM1 e 14,29% GSTM1 nulo (p = 0, 825). A GSTM1 e seu polimorfismo GSTM1 nulo não tiveram influência na resposta à quimioterapia pós-operatória. Dentre as pacientes que responderam 80,65% apresentavam GSTM1 e 19,35% nulo. Dentre as pacientes que não responderam, 87,50% apresentavam GSTM1 e 12,5% nulo ( p = 0,553). Conclusão: Não foi observada diferença na resposta ao tratamento com quimioterapia em pacientes com câncer avançado de ovário, em relação ao genótipo GSTM1 comparado ao seu polimorfismo GSTM1 nulo.

Interação entre consumo alimentar e polimorfismos da GSTM1 e GSTT1 no risco para o câncer de cabeça e pescoço: estudo caso-controle em São Paulo, Brasil / Interaction between dietary intake and GSTM1 and GSTT1 polymorphisms in head and neck cancer risk: a case-control study in São Paulo, Brazil

O objetivo foi investigar a interação entre fatores dietéticos e polimorfismos de enzimas de metabolização de xenobióticos (GSTM1 e GSTT1) associadas ao câncer de cabeça e pescoço em um estudo caso controle de base hospitalar, no Município de São Paulo, Brasil. Participaram 103 casos incidentes, histologicamente confirmados, e 101 controles. O consumo alimentar foi obtido por um questionário de frequência alimentar validado. Os polimorfismos GSTM1 e GSTT1 foram avaliados pelo método PCR. Observou-se aumento de risco no mais alto tercil de consumo de carne bovina na presença do alelo nulo da GSTM1 (OR = 10,79; IC95 por cento: 2,17-53,64) e GSTT1 (OR = 3,41; IC95 por cento: 0,43-27,21). Considerando-se a razão entre alimentos de origem animal e vegetal, verificou-se para o tercil intermediário a OR = 2,02 (IC95 por cento: 0,24-16,0) e no tercil superior OR = 3,23 (IC95 por cento: 0,40-25,92). Os resultados apontam para uma possível interação entre o consumo de carne e variantes polimórficas dos genes GSTM1 e GSTT1 na modulação do risco para o câncer de cabeça e pescoço, influenciados pelo consumo de alimentos de origem vegetal.

A hospital-based case-control study was conducted to investigate the potential interaction between dietary factors and polymorphisms in phase II metabolic enzymes GSTM1 and GSTT1, associated with head and neck cancer risk. The study included 103 histologically confirmed incident cases and 101 controls. Food intake was estimated with a validated food frequency questionnaire. The gene polymorphisms were evaluated by PCR. Increased risk was observed in the highest tertile of beef consumption in the presence of the GSTM1 (OR = 10.79; 95 percentCI: 2.17-53.64) and GSTT1 null alleles (OR = 3.41; 95 percentCI: 0.43-27.21). Assessment of dietary intake considering the ratio between animal product and vegetable consumption showed OR = 2.35 (95 percentCI: 0.27-19.85) in the intermediate tertile and OR = 3.36 (95 percentCI: 0.41-27.03) in the highest tertile. The results suggest a possible interaction between meat intake and GSTM1/GSTT1 polymorphisms in modulating the risk of head and neck cancer, influenced by vegetable consumption.

Preparation, identification and application of polyclonal antibody against major outer membrane protein of Chlamydia trachomatis E / 中华传染病杂志

Objective To express major outer membrane protein (MOMP) of Chlamydia trachomatis E and to prepare rabbit polyclonal antibody. Methods The recombinant plasmid MOMP/ pGEX6p-l prepared by our lab was introduced into E. coli. The protein was expressed and purified by gel recycling, then was injected into New Zealand rabbits to produce polyclonal antibodies. Enzyme linked immunosorbent assay (ELISA) was used to detect the titer of antibody. The antibody specificity was identified by Western blot and immunofluorescence. Results The fusion recombinant protein glutathione S-transferase (GST)-MOMP was successfully expressed in E. coli. The titer of antibody recombinant protein detected by Western blot and to the endogenous MOMP of Chlamydia trachomatis in vitro detected by immunofluorescence. Conclusions The recombinant MOMP is successfully expressed and the MOMP antibody with high titer and high specificity is obtained. which will be helpful for Chlamydia trachomatis detection and related clinical research.

Genetic polymorphism of GSTM1 to lung cancer patients in north Sichuan of China / 肿瘤研究与临床

Objective To analyze the genetic polymorphism of GSTM1 to lung cancer patients in north Sichuan of China and compare with race from other district.Methods PCR-based technique was used to detect the genotypes of GSTM1 in lung cancer patients.Results In local lung cancer patients,the frequency of homozygous deletions(null genotype) for GSTM1 was 58.4 % (73/125).Among the patients,the frequencys of null genotype for GSTM1 were 62.5 % (20/32) in female,56.9 % (53/93) in male,56.1% (32/57)in patients with squamous cell carcinoma and 54.8 % (17/31) in patients with adenocarcinoma,respectively.The frequency of deletions of GSTM1 in lung cancer patients from north Sichuan of China is slightly exceeding those of Europe and Americas (P ＜0.05) and similar to the domestic result (P ＞0.05).Conclusion The genetic polymorphism of GSTM1 to lung cancer patients in north Sichuan of China dosen' t show distinguished feature for this district and race.