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Evaluation of the expression levels of BRAFV600E mRNA in primary tumors of thyroid cancer using an ultrasensitive mutation assay.
Tran, Tien Viet; Dang, Kien Xuan; Pham, Quynh Huong; Nguyen, Ung Dinh; Trinh, Nhung Thi Trang; Hoang, Luong Van; Ho, Son Anh; Nguyen, Ba Van; Nguyen, Duc Trong; Trinh, Dung Tuan; Tran, Dung Ngoc; Orpana, Arto; Stenman, Ulf-Håkan; Stenman, Jakob; Ho, Tho Huu.
Afiliação
  • Tran TV; 103 Military Hospital, Vietnam Military Medical University, Hanoi, Vietnam.
  • Dang KX; Minerva Foundation Institute for Medical Research, Helsinki, Finland.
  • Pham QH; Department of Genomics and Cytogenetics, Institute of Biomedicine and Pharmacy (IBP), Vietnam Military Medical University, 222 Phung Hung street, Ha Dong district, Hanoi, Vietnam.
  • Nguyen UD; Department of Genomics and Cytogenetics, Institute of Biomedicine and Pharmacy (IBP), Vietnam Military Medical University, 222 Phung Hung street, Ha Dong district, Hanoi, Vietnam.
  • Trinh NTT; Department of Genomics and Cytogenetics, Institute of Biomedicine and Pharmacy (IBP), Vietnam Military Medical University, 222 Phung Hung street, Ha Dong district, Hanoi, Vietnam.
  • Hoang LV; Institute of Biomedicine and Pharmacy (IBP), Vietnam Military Medical University, Hanoi, Vietnam.
  • Ho SA; Institute of Biomedicine and Pharmacy (IBP), Vietnam Military Medical University, Hanoi, Vietnam.
  • Nguyen BV; Oncology Centre, 103 Military Hospital, Vietnam Military Medical University, Hanoi, Vietnam.
  • Nguyen DT; School of Medicine and Pharmacy, Vietnam National University, Hanoi, Vietnam.
  • Trinh DT; Pathology Department, 108 Military Central Hospital, Hanoi, Vietnam.
  • Tran DN; Department of Pathology, 103 Military Hospital, Vietnam Military Medical University, Hanoi, Vietnam.
  • Orpana A; Laboratory of Genetics, HUSLAB, Helsinki University Central Hospital, Helsinki, Finland.
  • Stenman UH; Department of Clinical Chemistry, Medicum, Helsinki University Hospital, University of Helsinki, Helsinki, Finland.
  • Stenman J; Minerva Foundation Institute for Medical Research, Helsinki, Finland.
  • Ho TH; Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.
BMC Cancer ; 20(1): 368, 2020 May 01.
Article em En | MEDLINE | ID: mdl-32357861
ABSTRACT

BACKGROUND:

The BRAFV600E gene encodes for the mutant BRAFV600E protein, which triggers downstream oncogenic signaling in thyroid cancer. Since most currently available methods have focused on detecting BRAFV600E mutations in tumor DNA, there is limited information about the level of BRAFV600E mRNA in primary tumors of thyroid cancer, and the diagnostic relevance of these RNA mutations is not known.

METHODS:

Sixty-two patients with thyroid cancer and non-malignant thyroid disease were included in the study. Armed with an ultrasensitive technique for mRNA-based mutation analysis based on a two step RT-qPCR method, we analysed the expression levels of the mutated BRAFV600E mRNA in formalin-fixed paraffin-embedded samples of thyroid tissues. Sanger sequencing for detection of BRAFV600E DNA was performed in parallel for comparison and normalization of BRAFV600E mRNA expression levels.

RESULTS:

The mRNA-based mutation detection assay enables detection of the BRAFV600E mRNA transcripts in a 10,000-fold excess of wildtype BRAF counterparts. While BRAFV600E mutations could be detected by Sanger sequencing in 13 out of 32 malignant thyroid cancer FFPE tissue samples, the mRNA-based assay detected mutations in additionally 5 cases, improving the detection rate from 40.6 to 56.3%. Furthermore, we observed a surprisingly large, 3-log variability, in the expression level of the BRAFV600E mRNA in FFPE samples of thyroid cancer tissue.

CONCLUSIONS:

The expression levels of BRAFV600E mRNA was characterized in the primary tumors of thyroid cancer using an ultrasensitive mRNA-based mutation assay. Our data inspires further studies on the prognostic and diagnostic relevance of the BRAFV600E mRNA levels as a molecular biomarker for the diagnosis and monitoring of various genetic and malignant diseases.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Mensageiro / Neoplasias da Glândula Tireoide / Carcinoma Papilar / Proteínas Proto-Oncogênicas B-raf / Câncer Papilífero da Tireoide / Mutação Tipo de estudo: Evaluation_studies / Observational_studies / Prognostic_studies Limite: Female / Humans / Male Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Mensageiro / Neoplasias da Glândula Tireoide / Carcinoma Papilar / Proteínas Proto-Oncogênicas B-raf / Câncer Papilífero da Tireoide / Mutação Tipo de estudo: Evaluation_studies / Observational_studies / Prognostic_studies Limite: Female / Humans / Male Idioma: En Ano de publicação: 2020 Tipo de documento: Article