Tumour-specific microRNA expression pattern in canine intestinal T-cell-lymphomas

Abstract

Intestinal T-cell lymphomas are common in dogs, but histopathological diagnosis remains challenging because of accompanying enteritis with lymphocyte involvement. Invasively taken full-layer biopsies are still required for reliable differentiation. The detection of specific microRNA expression patterns in canine intestinal T-cell lymphoma could provide new possibilities to differ intestinal lymphoma from benign inflammation and could lead to further understanding of lymphomagenesis. The objective of this study was to characterize microRNA expression in distinct groups of formalin-fixed and paraffin-embedded samples from canine intestinal T-cell lymphomas, lymphoplasmacellular enteritis and healthy intestinal tissue. In a preliminary test with two samples per group, total RNA was extracted (RNEasy FFPE Kit, Qiagen), reverse transcribed (miScript II RT Kit, Qiagen) and pre-amplified (miScript PreAmp PCR Kit, Qiagen). We performed comparative quantitative PCR on microRNA PCR Array plates (Qiagen) with pre-fabricated reactions for 183 different mature canine microRNAs. Subsequently, 12 microRNAs with conspicuous expression changes in the lymphoma group were selected and microRNA expression of all samples (n = 8) per group was analysed with individual microRNA assays (miScript Primer Assays, Qiagen) on the reverse transcribed RNA without pre-amplification. Our results revealed lymphoma-specific expression patterns, with down-regulation of the tumour-suppressing microRNAs miR-194, miR-192, miR-141 and miR-203, and up-regulation of oncogenic microRNAs, including microRNAs from the miR-106a~363 cluster. In addition, we detected only slight expression alterations between healthy intestinal tissue and lymphoplasmacellular enteritis cases. We conclude that microRNA expression patterns can be used to separate T-cell lymphomas from healthy tissue and benign inflammatory disorders.

Keywords: dog; lymphoma; microRNA; reverse transcriptase polymerase chain reaction.