Deregulated expression of Nucleophosmin 1 in gastric cancer and its clinicopathological implications

Background: the process of gastric carcinogenesis still remains to be elucidated. the identification of genes related to this process may help to reduce mortality rates through early diagnosis and the development of new anticancer therapies. Nucleophosmin 1 (NPM1) acts in ribosome biogenesis, centrosome duplication, maintenance of genomic stability, and embryonic development. Recently, NPM1 has been implicated in the tumorigenesis processes. Here, we evaluated NPM1 gene and protein expression in gastric tumors and in corresponding non-neoplastic gastric samples.Methods: NPM1 protein expression was determined by Western blot in 17 pairs of gastric tumors and corresponding non-neoplastic gastric tissue. the protein immunoreactivity was observed in 12 tumor samples. mRNA expression was evaluated by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in 22 pairs of gastric tumors and in matched non-neoplastic gastric tissue.Results: NPM1 protein expression was significantly reduced in gastric cancer samples compared to matched non-neoplastic gastric samples (P = 0.019). the protein level of NPM1 was reduced at least 1.5-fold in 35% of tumors compared to paired non-neoplastic gastric tissue. However, NPM1 immunoreactivity was detected in neoplastic and non-neoplastic cells, including in intestinal metaplastic, gastritis and inflammatory cells. NPM1 was mainly expressed in nucleus and nucleolus subcellular compartments. the staining intensity and the percentage of immunoreactive cells varied among the studied cases. the NPM1 mRNA level was reduced at least 1.5-fold in 45.5% of samples and increased in 27.3% of samples. An inverse correlation between protein and mRNA expression was detected (r = -0.509, P = 0.037). Intestinal-type gastric cancer presented higher mRNA levels than diffuse-type (P = 0.026). However, reduced NPM1 protein expression was associated with intestinal-type gastric cancer compared to matched non-neoplastic gastric samples (P = 0.018). in addition, tumors from patients with known distant metastasis presented reduced NPM1 protein levels compared to tumors from patients without distant metastasis (P < 0.001).Conclusion: Although the expression of NPM1 is heterogeneous in gastric tumors, our results suggest that NPM1 down-regulation may have a role in gastric carcinogenesis and may help in the selection of anticancer treatment strategies.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Morphol & Genet, Div Genet, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Orthoped & Traumatol, BR-04038031 São Paulo, BrazilUniv São Paulo, Sch Med, Dept Radiol, Expt Oncol Lab, BR-01246903 São Paulo, BrazilSão Paulo State Canc Inst, Ctr Translat Oncol, BR-01246000 São Paulo, BrazilFed Univ Para, Joao de Barros Barreto Univ Hosp, Oncol Res Ctr, BR-60673000 Belem, Para, BrazilFed Univ Para, Inst Biol Sci, Human Cytogenet Lab, BR-66073000 Belem, Para, BrazilUniversidade Federal de São Paulo, Dept Morphol & Genet, Div Genet, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Orthoped & Traumatol, BR-04038031 São Paulo, BrazilWeb of Scienc

Prohibitin Expression Deregulation in Gastric Cancer Is Associated with the 3 ' Untranslated Region 1630 C > T Polymorphism and Copy Number Variation

PHB is a reported oncogene and tumor suppressor in gastric cancer. Here, we evaluated whether the PHB copy number and the rs6917 polymorphism affect its expression in gastric cancer. Down-regulation and up-regulation of PHB were observed in the evaluated tumors. Reduced expression was associated with tumor dedifferentiation and cancer initiation. the T allele of the rs6917 polymorphism was associated with reduced PHB mRNA levels. Moreover, the up-regulation of PHB appeared to be regulated by the gain of additional gene copies. Thus, PHB copy number variation and differential expression of the rs6917 polymorphism may play a role in PHB transcriptional regulation.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Disciplina Genet, Dept Morfol & Genet, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Ortopedia & Traumatol, São Paulo, BrazilUniversidade Federal de São Paulo, Fac Med, Dept Radiol, Lab Oncol Expt, São Paulo, BrazilInst Canc Estado São Paulo, Ctr Invest Translat Oncol, São Paulo, BrazilFed Univ Para, Hosp Univ Joao Barros Barreto, BR-66059 Belem, Para, BrazilFed Univ Para, Inst Ciencias Biol, Lab Citogenet Humana, BR-66059 Belem, Para, BrazilUniversidade Federal de São Paulo, Disciplina Genet, Dept Morfol & Genet, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Ortopedia & Traumatol, São Paulo, BrazilUniversidade Federal de São Paulo, Fac Med, Dept Radiol, Lab Oncol Expt, São Paulo, BrazilWeb of Scienc

Quantitative expression analysis of genes related to aging and Alzheirmer’s Disease

Alzheimer Disease is the most common cause of dementia in elderly people. This disorder is characterized by a complex neurodegenerative process affecting multiple genes and proteins. Therefore, efforts have been made in identifying differentially expressed genes to be used as biomarkers in conditions such as aging and Alzheimer fs Disease. Purpose: Quantitative expression analysis of LR11, SNAP25 and SIRT1 genes, involved respectively in APP processing, synaptic transmission and neuroprotection, were investigated in brain and blood tissues. Furthermore, the expression between both tissues was also compared in order to find systemic markers that could represent the brain. Methods: mRNA quantification was performed using qRT-PCR in three post mortem brain regions (entorhinal and auditory cortices and hippocampus) of 10 Alzheimer fs Disease patients and 10 healthy elderly as well as in peripheral blood lymphocytes of 25 young, 20 healthy elderly and 35 Alzheimer fs Disease patients. We used the ƒ¢ƒ¢CT method in the analysis and the 2-ƒ¢ƒ¢CT formula to calculate the relative quantification. Results: LR11 mRNA quantification did not differ significantly concerning brain regions between Alzheimer fs Disease and healthy elderly subjects. However, in lymphocytes, its expression was threefold higher in Alzheimer fs Disease and healthy elderly subjects in comparison to the young group. Regarding SIRT1 gene, its expression was twofold higher in Alzheimer fs Disease patients than in healthy elderly for all brain regions. In lymphocytes, however, SIRT1 mRNA quantification was not significantly different among the three studied groups. Furthermore, lymphocytes showed a higher gene expression than brain regions for both LR11 and SIRT1 genes. In addition, a lack of SNAP25 expression was observed in blood lymphocytes. In brain tissues of the healthy elderly group, SNAP25 mRNA quantification was lower in the entorhinal cortex and hippocampus than in the auditory cortex. Furthermore, relative quantification of SNAP25 mRNA in brain regions of Alzheimer fs Disease patients was 34-39% of the total expression observed in the healthy elderly group. Conclusions: Our results presented some potential markers of Alzheimer fs Disease and aging processes. LR11 expression may be considered an age-related marker in blood tissue. The higher SIRT1 expression in Alzheimer fs Disease patients f brain suggested that this gene might be involved in compensatory mechanisms in the late stages of Alzheimer fs Disease. Both LR11 and SIRT1 brain expression was not correlated with blood expression, not being possible to consider them as systemic biomarker of the disease. Furthermore, it was not possible to use SNAP25 gene as a biomarker in blood once it was not expressed in this tissue. However, in the Alzheimer’s Disease patients’ brain, the decrease of SNAP25 expression might suggest an impairment of the synaptic function and neurotransmission found in the disease.A Doenca de Alzheimer e uma das causas mais comuns de demencia na populacao idosa. Esta doenca e caracterizada por um complexo processo neurodegenerativo que afeta diversos genes e proteinas. Desta forma, genes diferencialmente expressos tem sido investigados como possiveis biomarcadores para condicoes tais como o envelhecimento e a Doenca de Alzheimer. Objetivos: A expressao dos genes LR11, SNAP25 e SIRT1, envolvidos respectivamente com processamento da proteina precursora ƒÀ-Amiloide, transmissao sinaptica e neuroprotecao, foram quantificados em tecido cerebral e sanguineo. Alem disso, tambem foi comparada a expressao entre estes dois tecidos a fim de identificar marcadores sistemicos que pudessem estar correlacionados ao cerebro. Metodos: A quantificacao de RNAm foi realizada por meio de qRT-PCR em tres regioes cerebrais post mortem (cortices entorrinal e auditivo e hipocampo) de 10 pacientes com Doenca de Alzheimer e 10 idosos saudaveis, assim como em leucocitos de sangue periferico de 25 jovens, 20 idosos saudaveis e 35 pacientes com Doenca de Alzheimer. Foi utilizado o metodo ƒ¢ƒ¢CT e a formula 2-ƒ¢ƒ¢CT foi empregada no calculo da quantificacao relativa. Resultados: A quantificacao do RNAm de LR11 nao diferiu entre as regioes cerebrais de pacientes com Doenca de Alzheimer e de idosos saudaveis. Em leucocitos, porem, sua expressao foi cerca de tres vezes maior em pacientes com Doenca de Alzheimer e idosos saudaveis em relacao ao grupo de jovens. Em relacao ao gene SIRT1, sua expressao foi duas vezes maior em pacientes com Doenca de Alzheimer do que em idosos saudaveis para as tres regioes cerebrais estudadas. Em leucocitos, por sua vez, a quantificacao do RNAm de SIRT1 nao diferiu entre os tres grupos estudados. Alem disso, leucocitos de sangue periferico apresentaram maior expressao dos genes LR11 e SIRT1 quando comparados ao cerebro. Para o gene SNAP25, foi observada ausencia de expressao em leucocitos de sangue periferico. Em relacao ao tecido cerebral de idosos saudaveis, o cortex entorrinal e o hipocampo apresentaram menor expressao de SNAP25 do que o cortex auditivo. Alem disso, a quantificacao relativa de SNAP25 nas regioes cerebrais de pacientes com Doenca de Alzheimer variou entre 34 e 39% do total de expressao observada em idosos saudaveis. Conclusoes: Os resultados deste estudo indicaram alguns potenciais marcadores para o envelhecimento e para a Doenca de Alzheimer. O nivel de expressao de LR11 pode ser considerado um marcador de envelhecimento em tecido sanguineo. A maior expressao de SIRT1 no cerebro de pacientes com Doenca de Alzheimer sugere que este gene possa estar envolvido em mecanismos compensatórios nos estágios tardios da doença. A expressão cerebral dos genes LR11 e SIRT1 não se correlacionou com a de leucócitos, não sendo possível utilizá-los como biomarcadores sistêmicos da doença. O gene SNAP25 não se expressou em tecido sanguíneo na DA, desse modo, não foi indicado como biomarcador para a doença nesse tecido. No cérebro, porém, a diminuição de sua expressão em pacientes com DA sugere disfunção sináptica e de neurotransmissão associada à doença.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)TEDEBV UNIFESP: Teses e dissertaçõe

Apolipoprotein A1 gene polymorphisms as risk factors for hypertension and obesity

Several polymorphisms in apolipoprotein A1 (APOA1) gene have been associated with metabolic diseases. Increased transcription efficiency was observed in -75A allele carriers compared to -75G allele homozygotes. +83C allele was associated with higher body mass index and waist-to-hip ratio in type II diabetes subjects. -75G/A and +83C/T polymorphisms were analyzed by RFLP-PCR in 334 individuals from a Brazilian elderly cohort. APOA1 polymorphisms were associated with age-related morbidities, as well as with triglycerides, total cholesterol, HDL, VLDL, LDL, creatinine, urea, albumin, glycated hemoglobin and fasting glucose serum levels. Allele frequencies were 0.102 and 0.21, respectively, for -75A and +83T. -75G allele showed significant association with hypertension (P = 0.001). An association between +83C allele and obesity was observed (P = 0.040) and this allele also showed an association with hypertension in the presence of cardiovascular disease (P = 0.047). Moreover, +83T allele was associated with lower glycated hemoglobin values (P = 0.026). To our knowledge, there is no data associating this polymorphism with glycated hemoglobin. Furthermore, individuals carrying AT haplotype have lower risk for developing hypertension (P = 0.0002), while GT haplotype carriers present decreased risk to develop obesity comparing to GC haplotype (P = 0.025). APOA1 polymorphisms analysis may be a useful tool to identify risk factors for subjects and families and clarify the physiopathological role of these polymorphisms in age-related diseases, such as hypertension and obesity.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Escola Paulista Med, Dept Morfol & Genet, Disciplina Genet, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Med, Div Geriatr, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Morfol & Genet, Disciplina Genet, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Med, Div Geriatr, São Paulo, BrazilWeb of Scienc

Association of APOE, GCPII and MMP9 polymorphisms with common diseases and lipid levels in an older adult/elderly cohort

Background and aims: the characterization of candidate gene polymorphisms in elderly populations is an important tool for the identification of risk factors for age-related diseases and conditions. We aimed to genotype the APOE polymorphisms (rs429358 and rs7412), rs61886492 (1561C>T) and rs202720 of GCPII gene and rs3918242 (-1562C>T) of MMP9 gene in an older-adult/elderly cohort from Cuiaba city, Mato Grosso Brazil as well as to characterize risk factors for morbidities and conditions affecting this cohort.Methods: the studied population consisted of 570 subjects from Cuiaba city, Brazil, who were subjected to clinical interviews and blood collection for laboratory examinations and DNA extraction. Restriction Fragment Length Polymorphism Polymerase Chain Reaction (RFLP-PCR), sequence-specific primer PCR (SSP-PCR) and TaqMan (R) allelic discrimination assay were used for genotyping.Results: the frequencies of APOE epsilon 2 and epsilon 4 were 6.6% and 14.8%, respectively, and the frequencies of GCPII rs61886492 T allele, GCPII rs202720 C allele and MMP9 rs3918242 T allele were, respectively, 3.0%, 26.6% and 10.1%. Significant associations between APOE epsilon 2 allele with lower total cholesterol and LDL-cholesterol were found. in addition, MMP9 rs3918242 T allele was associated with higher LDL-cholesterol levels, suggesting a link between lipid metabolism alteration and cardiovascular disease.Conclusions: the present findings contributed to characterize risk factors specific for the studied population and to better understand the molecular physiopathology of common morbidities and conditions affecting older-adult/elderly people. (C) 2013 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Mato Grosso (FAPEMAT)Universidade Federal de São Paulo UNIFESP, Dept Morfol & Genet, Discipline Genet, BR-04023900 São Paulo, BrazilUniv Cuiaba, UNIC, Fac Med, Unidade Genet, Cuiaba, Mato Grosso, BrazilUniv Fed Mato Grosso, Fac Ciencias Med, Dept Ciencias Basicas Saude, Cuiaba, Mato Grosso, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Morfol & Genet, Discipline Genet, BR-04023900 São Paulo, BrazilWeb of Scienc