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S53G SNP in Cathepsin B: Can Be Used as a Molecular Biomarker to Breast Cancer?

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Open Journal of Genetics 2022年第4期12卷 43-56页

作者： Letícia Cristina Dalzy Castro danielle queiroz calcagno Carlos Eduardo de Melo Amaral Luiz Carlos Santana da Silva Programa de Pós-graduaç ã o em Oncologia e Ciências Médicas Universidade Federal do Pará Belém Brasil Laboratório de Erros Inatos do Metabolismo Universidade Federal do Pará Belém Brasil Núcleo de Pesquisa em Oncologia Universidade Federal do Pará Belém Brasil Fundaç ã o Centro de Hematologia e Hemoterapia do Pará Belém Brasil

Cathepsin B (CTS B) is a proteolytic enzyme that participates in several important biological processes. However, when it is altered can be involved in development of breast cancer, a disease with high incidence and mortality rate among women. Many studies have shown a correlation between high CTS B expression and breast tumor. Furthermore, it has been shown that the SNP rs1803250 (S53G) leads to structural and functional changes in protein that can make it pathogenic. The present study aimed to evaluate a possible association of SNP rs1803250 (S53G) with breast cancer. For this, real-time PCR was performed on a sample collected in the State of Pará, with 127 patients and 122 controls. The SNP frequency in this region was 0.12, according to a research project in progress that aims to identify Amerindians molecular alterations. This indicates that the SNP is found in region with a distribution close to worldwide frequency of 0.09. Our results showed that the SNP was in Hardy-Weinberg Equilibrium in collected sample, but C variant allelic frequency was 0.08 in both patients and control groups, which is extremely similar to global average. Moreover, homozygotes CC was not found in the sample and SNP genotypes frequency in patients and control groups was not significantly different. In addition, statistical analysis showed that the SNP did not have to correlate with tumor subtypes nor with tumor staging. Therefore, according to the analyzed sample, the SNP rs1803250 has no association with breast cancer and it cannot be considered a molecular biomarker for breast cancer.

关键词： Cathepsin B Polymorphism Breast Cancer

Clinical implication of 14-3-3 epsilon expression in gastric cancer

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World Journal of Gastroenterology 2012年第13期18卷 1531-1537页

作者： Mariana Ferreira Leal danielle queiroz calcagno Smia Demachki Paulo Pimentel Assumpo Roger Chammas Rommel Rodríguez Burbano Marília de Arruda Cardoso Smith Genetics Division Department of Morphology and GeneticFederal University of So PauloSo Paulo 04023-900Brazil Pathology Service Joo de Barros Barreto University HospitalFederal University of ParáBelém 60673-000Brazil Surgery Service Joo de Barros Barreto University HospitalFederal University of ParáBelém 60673-000Brazil Laboratory of Experimental Oncology School of MedicineUniversity of So PauloSo Paulo 01246-903Brazil Human Cytogenetics Laboratory Institute of Biological SciencesFederal University of ParáBelém 66073-000Brazil

AIM:To evaluate for the first time the protein and mRNA expression of 14-3-3εin gastric ***:14-3-3εprotein expression was determined by western blotting,and mRNA expression was examined by real-time quantitative RT-PCR in gastric tumors and their matched non-neoplastic gastric tissue ***:Authors observed a significant reduction of 14-3-3εprotein expression in gastric cancer(GC)samples compared to their matched non-neoplastic *** levels of 14-3-3εwere also associated with diffuse-type GC and early-onset of this *** data suggest that reduced 14-3-3εmay have a role in gastric carcinogenesis ***:Our results reveal that the reduced 14-3-3εexpression in GC and investigation of 14-3-3ε interaction partners may help to elucidate the carcino-genesis process.

关键词：胃癌临床意义 14-3-3蛋白 RT-PCR检测 mRNA表达量表肿瘤组织气相色谱仪

Interrelationship between chromosome 8 aneuploidy,C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma

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World Journal of Gastroenterology 2006年第38期12卷 6207-6211页

作者： danielle queiroz calcagno Mariana Ferreira Leal Aline Damaceno Seabra Andre Salim Khayat Elizabeth Suchi Chen Samia Demachki Paulo Pimentel Assumpcao Mario Henrique Girao Faria Silvia Helena Barem Rabenhorst Márcia Valéria Pitombeira Ferreira Marília de Arruda Cardoso Smith Rommel Rodríguez Burbano Human Cytogenetics and Toxicological Genetics Laboratory Department of BiologyCenter of Biological SciencesFederal University of ParáBelémPABrazil Genetics Division Department of Morphology Federal University of Sao PauloSao PauloSPBrazil Department of Pathology and Surgery Service Federal University of ParáBelémPABrazil Joao de Barros Barreto University Hospital Federal University of ParáBelémPABrazil Molecular Genetics Laboratory Department of Pathology Medical SchoolFederal University of CearáFortalezaCEBrazil

AIM: To investigate chromosome 8 numerical aberra- tions, C-MYC oncogene alterations and its expression in gastric cancer and to correlate these findings with histo- pathological characteristics of gastric tumors. METHODS: Specimens were collected surgically from seven patients with gastric adenocarcinomas. Immu- nostaining for C-MYC and dual-color fluorescence in situ hybridization (FISH) for C-MYC gene and chromosome 8 centromere were performed. RESULTS: All the cases showed chromosome 8 aneu- ploidy and C-MYC amplification, in both the diffuse and intestinal histopathological types of Lauren. No significant difference (P < 0.05) was observed between the level ofchromosome 8 ploidy and the site, stage or histological type of the adenocarcinomas. C-MYC high amplification, like homogeneously stained regions (HSRs) and double minutes (DMs), was observed only in the intestinal-type. Structural rearrangement of C-MYC, like translocation, was observed only in the diffuse type. Regarding C-MYC gene, a significant difference (P < 0.05) was observed between the two histological types. The C-MYC protein was expressed in all the studied cases. In the intestinal- type the C-MYC immunoreactivity was localized only in the nucleus and in the diffuse type in the nucleus and cytoplasm. CONCLUSION: Distinct patterns of alterations between intestinal and diffuse types of gastric tumors support the hypothesis that these types follow different genetic path- ways.

关键词： Chromosome 8 aneuploidy C-MYC amplification Immunostaining Gastric adenocarcinoma

MYC and gastric adenocarcinoma carcinogenesis

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World Journal of Gastroenterology 2008年第39期14卷 5962-5968页

作者： danielle queiroz calcagno Mariana Ferreira Leal Paulo Pimentel Assumpo Marília de Arruda Cardoso Smith Rommel Rodríguez Burbano Human Cytogenetics Laboratory Biology Science InstituteFederal University of ParáBelém PA 66075-900Brazil Genetics Division Department of Morphology and GeneticsFederal University of So PauloSo Paulo SP 04023-900Brazil Joo de Barros Barreto University Hospital Federal University of ParáBelém PA 66073-000Brazil

MYC is an oncogene involved in cell cycle regulation,cell growth arrest,cell adhesion,metabolism,ribosome biogenesis,protein synthesis,and mitochondrial function. It has been described as a key element of several carcinogenesis processes in humans. Many studies have shown an association between MYC deregulation and gastric cancer. MYC deregulation is also seen in gastric preneoplastic lesions and thus it may have a role in early gastric carcinogenesis. Several studies have suggested that amplification is the main mechanism of MYC deregulation in gastric cancer. In the present review,we focus on the deregulation of the MYC oncogene in gastric adenocarcinoma carcinogenesis,including its association with Helicobacter pylori (H pylori) and clinical applications.

关键词：胃癌肿瘤形成病理机制治疗方法

Identification of IL11RA and MELK amplification in gastric cancer by comprehensive genomic profiling of gastric cancer cell lines

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World Journal of Gastroenterology 2016年第43期22卷 9506-9514页

作者： danielle queiroz calcagno Sylvia Santomi Takeno Carolina Oliveira Gigek Mariana Ferreira Leal Fernanda Wisnieski Elizabeth Suchi Chen Taíssa Maíra Thomaz Araújo Eleonidas Moura Lima Maria Isabel Melaragno Samia Demachki Paulo Pimentel Assumpcao Rommel Rodriguez Burbano Marília Cardoso Smith Disciplina de Genética Departamento de Morfologia e GenéticaUniversidade Federal de Sao Paulo Núcleo de Pesquisas em Oncologia Hospital Universitário Joao de Barros Barreto Departamento de Biologia Molecular Universidade Federal da Paraíba Disciplina de Gastroenterologia Cirúrgica Universidade Federal de Sao Paulo Departamento de Ortopedia e Traumatologia Universidade Federal de Sao Paulo

AIM To identify common copy number alterations on gastric cancer cell *** Four gastric cancer cell lines(ACP02, ACP03, AGP01 and PG100) underwent chromosomal comparative genome hybridization and array comparative genome hybridization. We also confirmed the results by fluorescence in situ hybridization analysis using the bacterial artificial chromosome clone and quantitative real time PCR *** The amplification of 9p13.3 was detected in all cell lines by both methodologies. An increase in the copy number of 9p13.3 was also confirmed by fluorescence in situ hybridization analysis. Moreover, the interleukin 11 receptor alpha(IL11RA) and maternal embryonic leucine zipper kinase(MELK) genes, which are present in the 9p13.3 amplicon, revealed gains of the MELK gene in all the cell lines studied. Additionally, a gain in the copy number of IL11 RA and MELK was observed in 19.1%(13/68) and 55.9%(38/68) of primary gastric adenocarcinoma samples, respectively. CONCLUSION The characterization of a small gain region at 9p13.3 in gastric cancer cell lines and primary gastric adenocarcinoma samples has revealed MELK as a candidate target gene that is possibly related to the development of gastric cancer.

关键词： IL11RA Gastric cancer Genomic profiling MELK 9p13.3

DNA and histone methylation in gastric carcinogenesis

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World Journal of Gastroenterology 2013年第8期19卷 1182-1192页

作者： danielle queiroz calcagno Carolina Oliveira Gigek Elizabeth Suchi Chen Rommel Rodriguez Burbano Marília de Arruda Cardoso Smith Department of Morphology and Genetics Federal University of Sao PauloSao PauloSP 04023-900Brazil Laboratory of Human Cytogenetics Institute of Biological SciencesFederal University of ParáBelémPA 66075-110Brazil

Epigenetic alterations contribute significantly to the development and progression of gastric cancer,one of the leading causes of cancer death *** refers to the number of modifications of the chromatin structure that affect gene expression without altering the primary sequence of DNA,and these changes lead to transcriptional activation or silencing of the *** the years,the study of epigenetic processes has increased,and novel therapeutic approaches that target DNA methylation and histone modifications have emerged.A greater understanding of epigenetics and the therapeutic potential of manipulating these processes is necessary for gastric cancer ***,we review recent research on the effects of aberrant DNA and histone methylation on the onset and progression of gastric tumors and the development of compounds that target enzymes that regulate the epigenome.

关键词： Epigenetic DNA methylation Histone methylation Gastric cancer Gastric carcinogenesis

Promoter hypermethylation of CDH1, FHIT, MTAP and PLAGL1 in gastric adenocarcinoma in individuals from Northern Brazil

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World Journal of Gastroenterology 2007年第18期13卷 2568-2574页

作者： Mariana Ferreira Leal Eleonidas Moura Lima Patrícia Natália Oliveira Silva Paulo Pimentel Assumpo danielle queiroz calcagno Spencer Luiz Marques Payo Rommel Rodríguez Burbano Marília de Arruda Cardoso Smith Genetics Division Department of Morphology Federal University of So Paulo So Paulo SP Brazil Department of Biology Campus Ministro Reis Velloso/Parnába Federal University of Piauí PI Brazil Joo de Barros Barreto University Hospital Federal University of Pará Belém PA Brazil Human Cytogenetics and Toxicological Genetics Laboratory Department of Biology Center of Biological Sciences Federal University of Pará Belém PA Brazil Genetics and Molecular Biology Hemocenter Faculty of Medicine of Marília Marília SP Brazil

AIM: To evaluate the methylation status of CDH1, FHIT, MTAP and PLAGL1 promoters and the association of these findings with clinico-pathological characteristics. METHODS: Methylation-specific PCR (MSP) assay was performed in 13 nonneoplastic gastric adenocarcinoma, 30 intestinal-type gastric adenocarcinoma and 35 diffuse- type gastric adenocarcinoma samples from individuals in Northern Brazil. Statistical analyses were performed using the chi-square or Fisher’s exact test to assess associations between methylation status and clinico- pathological characteristics. RESULTS: Hypermethylation frequencies of CDH1, FHIT, MTAP and PLAGL1 promoter were 98.7%, 53.9%, 23.1% and 29.5%, respectively. Hypermethylation of three or four genes revealed a significant association with diffuse-type gastric cancer compared with nonneoplastic cancer. A higher hypermethylation frequency wassignificantly associated with H pylori infection in gastric cancers, especially with diffuse-type. Cancer samples without lymph node metastasis showed a higher FHIT hypermethylation frequency. MTAP hypermethylation was associated with H pylori in gastric cancer samples, as well as with diffuse-type compared with intestinal-type. In diffuse-type, MTAP hypermethylation was associated with female gender. CONCLUSION: Our findings show differential gene methylation in tumoral tissue, which allows us to conclude that hypermethylation is associated with gastric carcinogenesis. MTAP promoter hypermethylation can be characterized as a marker of diffuse-type gastric cancer, especially in women and may help in diagnosis, prognosis and therapies. The H pylori infectious agent was present in 44.9% of the samples. This infection may be correlated with the carcinogenic process through the gene promoter hypermethylation, especially the MTAP promoter in diffuse-type. A higher H pylori infection in diffuse-type may be due to greater genetic predisposition.

关键词：巴西北部地区胃腺癌 CDH1 FHIT MTAP PLAGL1 启动子 DNA超甲基化

Traps and trumps from adjacent-to-tumor samples in gastric cancer research

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Chinese Journal of Cancer Research 2018年第5期30卷 564-567页

作者： Paulo Pimentel de Assumpcao Andre Salim Khayat Taissa Maira Thomaz Araujo Williams Fernandes Barra Geraldo Ishak Mine Maria Pereira Cruz Ramos Sidney Emanuel Batista dos Santos Andrea Kely Campos Ribeiro dos Santos Samia Demachki Paula Barauna de Assumpcao danielle queiroz calcagno Ney Pereira Carneiro dos Santos Monica Barauna de Assumpcao Fabiano Cordeiro Moreira Andre Mauricio Ribeiro dos Santos Carolina Barauna de Assumpcao Gregory Joseph Riggins Rommel Mario Rodriguez Burbano Oncology Research Center Federal University of Para Belem 66075-110 Brazil PhD Program in Genetics and Molecular Biology Federal University of Para Belem 66075-110 Brazil Gastrointestinal Endoscopy Service Joao de Barros Barreto University Hospital Belem 66073-000 Brazil Department of Surgical Oncology Oswaldo Cruz Germany Hospital Sao Paulo 01327-001 Brazil Department of Neurosurgery Johns Hopkins University Baltimore 21287 USA Laboratory of Molecular Biology Ophir Loyola Hospital Belem 66060-281 Brazil

The search for cancer biomarkers is frequently based on comparisons between tumors and adjacent-to-tumor samples. However, even after histological confirmation of been free of cancer cells, these adjacent-to-tumor samples might harbor molecular alterations which are not sufficient to cause them to look like cancer, but can differentiate these cells from normal cells. When comparing them, potential biomarkers are missed, and mainly the opportunity of finding initial aberrations presents in both tumors and adjacent samples, but not in true normal samples from non-cancer patients, resulting in misinterpretations about the carcinogenic process. Nevertheless, collecting adjacent-to-tumor samples brings trumps to be explored. The addition of samples from non-cancer patients opens an opportunity to increase the finds of the molecular cascade of events in the carcinogenic process. Differences between normal samples and adjacent samples might represent the first steps of the carcinogenic process. Adding samples of non-cancer patients to the analysis of molecular alterations relevant to the carcinogenic process opens a new window of opporttmides to the discovery of cancer biomarkers and molecular targets.

关键词： Adjacent-to-tumor trumps traps

Reference genes for quantitative RT-PCR data in gastric tissues and cell lines

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World Journal of Gastroenterology 2013年第41期19卷 7121-7128页

作者： Fernanda Wisnieski danielle queiroz calcagno Mariana Ferreira Leal Leonardo Caires dos Santos Carolina de Oliveira Gigek Elizabeth Suchi Chen Thaís Brilhante Pontes Paulo Pimentel Assumpo Mnica Barauna de Assumpo Smia Demachki Rommel Rodríguez Burbano Marília de Arruda Cardoso Smith Disciplina de Genética Departamento de Morfologia e GenéticaUniversidade Federal de So PauloSo Paulo 04023900Brazil Departamento de Ortopedia e Traumatologia Universidade Federal de So PauloSo Paulo 04038031Brazil Núcleo de Pesquisa em Oncologia Hospital Joo de Barros BarretoUniversidade Federal do ParáBelém 60673000Brazil Laboratório de Citogenética Humana Instituto de Ciências BiológicasUniversidade Federal do ParáBelém 66075110Brazil

AIM:To evaluate the suitability of reference genes in gastric tissue samples and cell ***:The suitability of genes ACTB,B2M,GAPDH,RPL29,and 18S rRNA was assessed in21 matched pairs of neoplastic and adjacent nonneoplastic gastric tissues from patients with gastric adenocarcinoma,27 normal gastric tissues from patients without cancer,and 4 cell lines using reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR).The ranking of the best single and combination of reference genes was determined by NormFinder,geNorm,BestKeeper,and *** addition,GenEx software was used to determine the optimal number of reference *** validate the results,the mRNA expression of a target gene,DNMT1,was quantified using the different reference gene combinations suggested by the various software packages for ***:ACTB was the best reference gene for all gastric tissues,cell lines and all gastric tissues plus cell ***+B2M or ACTB+B2M was the best combination of reference genes for all the gastric *** the other hand,ACTB+B2M was the best combination for all the cell lines tested and was also the best combination for analyses involving all the gastric tissues plus cell *** to the GenEx software,2 or 3 genes were the optimal number of references genes for all the gastric *** relative quantification of DNMT1 showed similar patterns when normalized by each combination of reference *** level of expression of DNMT1 in neoplastic,adjacent non-neoplastic and normal gastric tissues did not differ when these samples were normalized using GAPDH+B2M(P=0.32),ACTB+B2M(P=0.61),or GAPDH+B2M+ACTB(P=0.44).CONCLUSION:GAPDH+B2M or ACTB+B2M is the best combination of reference gene for all the gastric tissues,and ACTB+B2M is the best combination for the cell lines tested.

关键词： Gastric cancer Reference gene Normalization Gene expression Quantitative real-time polymerase chain reaction