21 research outputs found
Polymorphism Analysis of Genes Involved in Xenobiotic Metabolism and Circadian Rhythm in Human Breast Cancer
Get PDFIndividual response to xenobiotic exposures depends on the dynamics of xenobiotic metabolism and the circadian clock system, among other factors. Since these systems are closely related, polymorphisms in their key genes may have an impact on how carcinogenic compounds are metabolized, and therefore on the risk of tumor development. Whereas the mammary gland is exposed to agents that damage DNA, it was considered of high interest to study xenobiotic metabolizing genes (XMG) and clock genes in this tissue. Our aim was to analyze genotype and allele frequencies of polymorphisms in the XMG N-acetyl transferase 2 (NAT2) and glutathione S-transferase T1 (GSTT1), and in the clock genes period 3 (PER3) and CLOCK in human breast tumor samples. As well, it was if these polymorphisms were in linkage disequilibrium. 65 samples were genotyped for polymorphisms in GSTT1 (null), NAT2 (C481T, G590A and G857A), CLOCK (T3111C) and PER3 (length polymorphism), by PCR and PCR-RFLP. For GSTT1, 20% of the samples showed total absence of the gene. When NAT2 genotypes were grouped by their associated acetylator phenotype, 5% of rapid, 49% of intermediate and 46% of slow acetylator phenotypes were indicated. Allele frequencies for CLOCK were T=0.78 and C=0.22; for PER3, they were 0.66 for the 4-repeats allele and 0.34 for the 5-repeats allele. Linkage disequilibrium test indicated evidence of strong linkage between NAT2 and CLOCK (χ2=13.076; p=0.005). With regard to allele and genotype frequencies, our results are in agreement with those reported for similar populations. The evidence of linkage disequilibrium in our breast cancer samples is interesting and requires further investigation. This work constitutes a first approximation to a combined study of polymorphisms in XMG and clock genes in breast cancer Argentinian patients. Future studies will attempt to address the role of these and other polymorphisms in cancer risk, prognosis and response to treatment.Fil: Cerliani, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Richard, Silvina Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentin
Polymorphism Analysis of Genes Involved in Xenobiotic Metabolism and Circadian Rhythm in Human Breast Cancer
Get PDFIndividual response to xenobiotic exposures depends on the dynamics of xenobiotic metabolism and the circadian clock system, among other factors. Since these systems are closely related, polymorphisms in their key genes may have an impact on how carcinogenic compounds are metabolized, and therefore on the risk of tumor development. Whereas the mammary gland is exposed to agents that damage DNA, it was considered of high interest to study xenobiotic metabolizing genes (XMG) and clock genes in this tissue. Our aim was to analyze genotype and allele frequencies of polymorphisms in the XMG N-acetyl transferase 2 (NAT2) and glutathione S-transferase T1 (GSTT1), and in the clock genes period 3 (PER3) and CLOCK in human breast tumor samples. As well, it was if these polymorphisms were in linkage disequilibrium. 65 samples were genotyped for polymorphisms in GSTT1 (null), NAT2 (C481T, G590A and G857A), CLOCK (T3111C) and PER3 (length polymorphism), by PCR and PCR-RFLP. For GSTT1, 20% of the samples showed total absence of the gene. When NAT2 genotypes were grouped by their associated acetylator phenotype, 5% of rapid, 49% of intermediate and 46% of slow acetylator phenotypes were indicated. Allele frequencies for CLOCK were T=0.78 and C=0.22; for PER3, they were 0.66 for the 4-repeats allele and 0.34 for the 5-repeats allele. Linkage disequilibrium test indicated evidence of strong linkage between NAT2 and CLOCK (χ2=13.076; p=0.005). With regard to allele and genotype frequencies, our results are in agreement with those reported for similar populations. The evidence of linkage disequilibrium in our breast cancer samples is interesting and requires further investigation. This work constitutes a first approximation to a combined study of polymorphisms in XMG and clock genes in breast cancer Argentinian patients. Future studies will attempt to address the role of these and other polymorphisms in cancer risk, prognosis and response to treatment.Instituto Multidisciplinario de Biología Celula
Sensitization to oxaliplatin in HCT116 and HT29 cell lines by metformin and ribavirin and differences in response to mitochondrial glutaminase inhibition
Get PDFAim of study: In the present study, we evaluated the effect of ribavirin and metformin on the sensitivity of oxaliplatin and 5-fluorouracil (5-FU) on colon cancer.
Materials and Methods: Cell viability of two commercially available colon cancer cell lines (HT29 and HCT116) were analyzed by sulforhodamine B (SRB) assay.
Results: A clinically achievable and nontoxic concentration of ribavirin and metformin showed a significant synergistic effect on oxaliplatin in HT29 and HCT116 cell lines. Ribavirin showed a synergistic effect on oxaliplatin in HT29 (R = 2.93, P < 0.001) and HCT116 (R = 1.71, P < 0.001), while only in HT29 metformin synergized with oxaliplatin by 2.66 (± 0.28, P < 0.01). In addition, both cell lines showed significant differences in response to Compound 968, inhibitor of mitochondrial glutaminase activity.
Conclusion: The data suggested that these cell lines not only turn to metabolic different sustainability process after oxaliplatin treatment but that they also have different basal metabolic requirements of glutamine in vitro which can be exploits in the future for colorectal cancer (CRC) treatment and further studies are required.Instituto Multidisciplinario de Biología Celula
Sensitization to oxaliplatin in HCT116 and HT29 cell lines by metformin and ribavirin and differences in response to mitochondrial glutaminase inhibition
Get PDFAim of study: In the present study, we evaluated the effect of ribavirin and metformin on the sensitivity of oxaliplatin and 5-fluorouracil (5-FU) on colon cancer.
Materials and Methods: Cell viability of two commercially available colon cancer cell lines (HT29 and HCT116) were analyzed by sulforhodamine B (SRB) assay.
Results: A clinically achievable and nontoxic concentration of ribavirin and metformin showed a significant synergistic effect on oxaliplatin in HT29 and HCT116 cell lines. Ribavirin showed a synergistic effect on oxaliplatin in HT29 (R = 2.93, P < 0.001) and HCT116 (R = 1.71, P < 0.001), while only in HT29 metformin synergized with oxaliplatin by 2.66 (± 0.28, P < 0.01). In addition, both cell lines showed significant differences in response to Compound 968, inhibitor of mitochondrial glutaminase activity.
Conclusion: The data suggested that these cell lines not only turn to metabolic different sustainability process after oxaliplatin treatment but that they also have different basal metabolic requirements of glutamine in vitro which can be exploits in the future for colorectal cancer (CRC) treatment and further studies are required.Instituto Multidisciplinario de Biología Celula
Analysis association between mitochondrial genome instability and xenobiotic metabolizing genes in human breast cancer
Get PDFThe aim of this study was to determine the existence of association between the genetic polymorphisms of metabolizing genes GSTM-1, GSTT-1,and NAT-2,and the presence of mitochondrial genome instability (mtGI) in breast cancer cases. Ninety-four pairs of tumoral/nontumoral breast cancer samples were analyzed. Our samples showed 40.42% of mtGI by analysis of two D-loop region markers, a (CA)n mtMS starting at the 514-bp position, and four informative MnlI sites between the 16, 108-16, 420-bp. GSTM-1null genotype has shown a significant association with mtGI presence (χ2 = 7.62; P= 0.006) in breast cancer cases; moreover, these genotypes also are related to an increased risk for mtDNA damage (odds ratio (OR) = 3.71 (1.41-9.88); 95% Cornfield confidence interval (CI)). These results suggest that the absence of GSTM-1enzymatic activity favors chemical actions in damaging the mtDNA. Analysis of GSTT-1and NAT-2polymorphisms showed no association with mtGI (χ2 = 0.03; P = 0.87 and χ2 = 2.76; P = 0.09, respectively). The analysis of invasive breast cancer cases showed mtGI in 74.36% of ILC cases (29 of 39 samples), and in only 18.75% (9 out of 48) IDC cases; this result suggests a possible relation between mtDNA mutations and variations in molecular pathways of tumor development.Instituto Multidisciplinario de Biología CelularFacultad de Ciencias Médica
Association between PER3 length polymorphism and onco-hematological diseases and its influence on patients functionality
Get PDFCircadian clock gene PER3 and its length polymorphism may have a role in oncogenesis as clock genes act as key regulators of cell cycle and DNA repair pathways. The polymorphism may affect the condition of patients who show disrupted circadian rhythm due to tumor development. The aim was to assess the association between PER3 polymorphism and onco-hematological diseases, and analyze whether this variant has an impact on patient’s functionality.
We conducted a case-control study on 125 patients with onco-hematological diseases and 310 control patients. PER3 allelic variants were detected by using polymerase chain reaction. Sociodemographic data and information on patient’s habits and functionality were obtained through questionnaire. Genotypes 4/5 + 5/5 showed an odd ratio (OR) = 1.39, with no statistical significance. However, those genotypes were associated with a two-fold increase in the risk of acute/chronic lymphoblastic/myeloblastic leukemia, taken all together. The occurrence of “changes in humor during last two months” was significantly associated with onco-hematological diseases. “Fatigue on awakening” and “self-reported snore” were associated with cases carrying the 4/5 or 5/5 genotypes. The results suggested that PER3 polymorphism may have a role in the risk of leukemia, and might be a possible marker for individual differences in susceptibility to sleep disruption. This work provides insights for the identification of individuals at high risk of cancer, and those who are more susceptible to circadian disruption, which may decrease the physiological defenses against the tumor
Cigarette smoking, dietary habits and genetic polymorphisms in <i>GSTT1</i>, <i>GSTM1</i> and <i>CYP1A1</i> metabolic genes: A case-control study in oncohematological diseases
Get PDFAim: To analyze the association between oncohematological diseases and GSTT1 /GSTM1 /CYP1A1 polymorphisms, dietary habits and smoking, in an argentine hospitalbased case-control study.
Methods: This hospital-based case-control study involved 125 patients with oncohematological diseases and 310 control subjects. A questionnaire was used to obtain sociodemographic data and information about habits. Blood samples were collected, and DNA was extracted using salting out methods. Deletions in GSTT1 and GSTM1 (null genotypes) were addressed by PCR. CYP1A1 MspI polymorphism was detected by PCR-RFLP. Odds ratio (OR) and 95%CI were calculated to estimate the association between each variable studied and oncohematological disease.
Results: Women showed lower risk of disease compared to men (OR 0.52, 95%CI: 0.34-0.82, P = 0.003). Higher levels of education (> 12 years) were significantly associated with an increased risk, compared to complete primary school or less (OR 3.68, 95%CI: 1.82-7.40, P GSTT1, GSTM1 and CYP1A1 polymorphisms showed no significant association with oncohematological diseases. When analyzing the interaction between polymorphisms and tobacco smoking or dietary habits, no statistically significant associations that modify disease risk were found.
Conclusion: We reported an increased risk of oncohematological diseases associated with meat and coffee intake. We did not find significant associations between genetic polymorphisms and blood cancer.Instituto Multidisciplinario de Biología Celula
Association between PER3 length polymorphism and onco-hematological diseases and its influence on patients’ functionality
Get PDFCircadian clock gene PER3 and its length polymorphism may have a role in oncogenesis as clock genes act as key regulators of cell cycle and DNA repair pathways. The polymorphism may affect the condition of patients who show disrupted circadian rhythm due to tumor development. The aim was to assess the association between PER3 polymorphism and onco-hematological diseases, and analyze whether this variant has an impact on patient’s functionality. We conducted a case-control study on 125 patients with onco-hematological diseases and 310 control patients. PER3 allelic variants were detected by using polymerase chain reaction. Sociodemographic data and information on patient’s habits and functionality were obtained through questionnaire. Genotypes 4/5 + 5/5 showed an odd ratio (OR) = 1.39, with no statistical significance. However, those genotypes were associated with a two-fold increase in the risk of acute/chronic lymphoblastic/myeloblastic leukemia, taken all together. The occurrence of “changes in humor during last two months” was significantly associated with onco-hematological diseases. “Fatigue on awakening” and “self-reported snore” were associated with cases carrying the 4/5 or 5/5 genotypes. The results suggested that PER3 polymorphism may have a role in the risk of leukemia, and might be a possible marker for individual differences in susceptibility to sleep disruption. This work provides insights for the identification of individuals at high risk of cancer, and those who are more susceptible to circadian disruption, which may decrease the physiological defenses against the tumor.Fil: Cerliani, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Gili, Juan Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET; ArgentinaFil: Pavicic, Walter Hernan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Klein, Graciela Eleonor. Gobierno de la Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal General de Agudos "prof. Dr. Rodolfo Rossi".; ArgentinaFil: Saba, Silvia. Gobierno de la Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal General de Agudos "prof. Dr. Rodolfo Rossi".; ArgentinaFil: Richard, Silvina Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentin
Cigarette smoking, dietary habits and genetic polymorphisms in <i>GSTT1</i>, <i>GSTM1</i> and <i>CYP1A1</i> metabolic genes: A case-control study in oncohematological diseases
Get PDFAim: To analyze the association between oncohematological diseases and GSTT1 /GSTM1 /CYP1A1 polymorphisms, dietary habits and smoking, in an argentine hospitalbased case-control study.
Methods: This hospital-based case-control study involved 125 patients with oncohematological diseases and 310 control subjects. A questionnaire was used to obtain sociodemographic data and information about habits. Blood samples were collected, and DNA was extracted using salting out methods. Deletions in GSTT1 and GSTM1 (null genotypes) were addressed by PCR. CYP1A1 MspI polymorphism was detected by PCR-RFLP. Odds ratio (OR) and 95%CI were calculated to estimate the association between each variable studied and oncohematological disease.
Results: Women showed lower risk of disease compared to men (OR 0.52, 95%CI: 0.34-0.82, P = 0.003). Higher levels of education (> 12 years) were significantly associated with an increased risk, compared to complete primary school or less (OR 3.68, 95%CI: 1.82-7.40, P GSTT1, GSTM1 and CYP1A1 polymorphisms showed no significant association with oncohematological diseases. When analyzing the interaction between polymorphisms and tobacco smoking or dietary habits, no statistically significant associations that modify disease risk were found.
Conclusion: We reported an increased risk of oncohematological diseases associated with meat and coffee intake. We did not find significant associations between genetic polymorphisms and blood cancer.Instituto Multidisciplinario de Biología Celula
Global DNA Methylation levels analysis in a serie of Hematological, Breast and Colorectal cancer samples from Argentina
Get PDFUnlike their normal counterparts, tumor cells exhibited highly variable CpG methylation levels in a large proportion of the genome, which can lead to malignant cell transformation through multiple pathways. This prompted us to assess the extent of LINE1 methylation, a surrogate marker of global DNA methylation, of samples derived from controls and cancer patients from Argentina. Preliminary DNA methylation results from selected samples were replicated in a large serie of 146 controls (blood) and various cancer types: 112 oncohematological cancer (HemCa), 70 colorectal cancer (CRC) and 68 breast cancer (BrCa) samples. Further, we evaluated correlation with biological, clinical and demographic features. Blood samples were available in all cases, and for solid tumors paired tumoral/non-tumoral adjacent tissues (T/N) were available too. LINE1 methylation level was analyzed by MS-MLPA method. HemCa cases showed statistically significant higher LINE1 methylation level (p>0.001) compared to controls (mean 0.93 and 0.84, respectively). This variation could be a consequence of chemotherapy. Methylation status in blood (0.86) and N tissue (0.87) from BrCa cases did not differ from controls, while levels in T tissue (0.88) were significantly higher than controls (p<0.05). No differences between N and T tissues were found. CRC cases showed hypomethylation for LINE1 when comparing T (0.81) to blood (0.87) or N tissues (0.88), reaching statistical significance of p<0.05 and p<0.001, respectively. This is in line with reported results. We found a negative correlation between individual age and methylation level in controls (-0.17, p=0.04), and BrCa T tissue (-0.33, p=0.03). Finally, no relevant associations between global methylation and mitochondrial genome variation (copy number and ancestry) were found for controls and HemCa sample sets. LINE1 methylation analysis in samples from lung, ovarian, pancreatic and skin cancers are ongoing.Fil: Cerliani, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Mayordomo, Andrea Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Sanchez Dova, Anaclara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Piñero, Tamara Alejandra. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaFil: Cajal, Andrea. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaFil: Jauk Vitali, Federico. Hospital Italiano; ArgentinaFil: Garcia Rivello, Hernan Jorge. Hospital Italiano; ArgentinaFil: Vaccaro, Carlos Alberto. Hospital Italiano; ArgentinaFil: Richard, Silvina Mariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Bravi, Claudio Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Pavicic, Walter Hernan. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaReunión Anual de Sociedades de BiocienciaArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de Farmacología ExperimentalSociedad Argentina de BiologíaSociedad Argentina de BiologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de Laboratori
