A simple method for RNA isolation from formalin-fixed and paraffin-embedded lymphatic tissues

• Published in: Experimental and molecular pathology 2003-06, Vol.74 (3), p.336-340
• Main Authors: Körbler, Tajana, Grs̆ković, Marica, Dominis, Marija, Antica, Mariastefania
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; DNA Primers - chemistry; Electrophoresis, Agar Gel; Fixatives; Formaldehyde; Formalin fixed; Hodgkin Disease - genetics; Hodgkin Disease - metabolism; Hodgkin Disease - pathology; Humans; Investigative techniques, diagnostic techniques (general aspects); Lymph nodes; Lymph Nodes - chemistry; Lymph Nodes - pathology; Lymphoma, Non-Hodgkin - genetics; Lymphoma, Non-Hodgkin - metabolism; Lymphoma, Non-Hodgkin - pathology; Medical sciences; Miscellaneous. Technology; Paraffin embedded; Paraffin Embedding; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA isolation; RNA, Messenger - metabolism; RNA, Neoplasm - isolation & purification; Transcription factor; Transcription Factors
• ISSN: 0014-4800; 1096-0945
• DOI: 10.1016/S0014-4800(03)00024-8

Gene activation that lies beneath lymphoid cell differentiation has been one of the most explored issues in immunology in the recent years. However, the analysis of this molecular event in lymphoproliferative diseases is often hampered by the lack of fresh material. Most tissues available for routine histological investigation are formalin fixed and paraffin embedded. Gene expression in such specimens could be analyzed using reverse transcription of mRNA and the polymerase chain reaction (RT-PCR). Therefore we adjusted and established a method for mRNA isolation from such specimens by a combination of previously reported protocols and a modification of the phenol/chloroform extraction method. Given the significance of transcription factors in the human hemopoietic system, we investigated whether mRNA could be successfully isolated from archival tissue for a study on expression of Ikaros family transcription factors in lymphatic tissue. Although quantitative analysis of RNA isolated from archival tissue is probably not feasible due to the unpredictable degree of RNA isolation varying from sample to sample, we show here that screening analysis is possible and simple.