Microdissecˋˋo e captura a laser na investigaˋˋo do gene TP53 em tecidos inclu赤dos em parafina

作者：Ihlaseh; Shadia Muhammad; Oliveira; Maria Luiza Cotrim Sartor de; Silva; Glenda Nicioli da; Franchi; Carla Adriene da Silva; Camargo; Joo Lauro Viana de

来源：Jornal Brasileiro de Patologia e Medicina Laboratorial, 2007.

DOI：10.1590/S1676-24442007000100012

• laser-capture microdissection
• dna extraction
• nested-pcr
• tp53 gene
• ciclophilin gene
• paraffin-embedded tissues.

backgorund: laser-capture micro-dissection (lcm) is a recently developed procedure that provides single cells or specific cell groups for molecular analysis. currently, there are few lcm systems in brazil, in such a way that it is necessary to disseminate the technical procedures inherent to the methodology, and also to characterize its enormous potential for diagnosis and research. objective: this study describes the standardization of lcm and dna extraction from formalin fixed and paraffin-embedded tissues. material and method: the gene tp53 exon 8 and the cyclophilin gene were studied in normal and neoplastic liver and kidney samples from a chemical carcinogenesis model in rat. dna extraction was confirmed by nested-pcr. results: histological sections preparation for lcm and the nested-pcr procedures were standardized; 48.3% amplifications of the gene tp53 exon 8 and 51.7% of the cyclophilin gene samples were obtained. when at least one of the gene segments was considered, 79.3% samples presented amplification. discussion and conclusion: procedures for dna extraction from formalin fixed and paraffin-embedded tissues collected by lcm were standardized. they can be useful for dna collection for molecular studies.