Molecular and epigenetic analysis of the fragile histidine triad tumour suppressor gene in equine sarcoids
- Equal contributors
1 Institute for Animal Production System in Mediterranean Environment, National Research Council, Via Argine, 1085, 80147 Naples, Italy
2 Department of Pathology and Animal Health, University of Naples Federico II, Via Veterinaria, 1, 80137 Naples, Italy
3 Institute of Genetic and Biophysics ABT, National Research Council, Via P. Castellino 111, 80131 Naples, Italy
4 IRCCS Neuromed, Pozzilli, Italy
BMC Veterinary Research 2012, 8:30 doi:10.1186/1746-6148-8-30Published: 16 March 2012
Sarcoids are peculiar equine benign tumours. Their onset is associated with Bovine Papillomavirus type -1 or -2 (BPV-1/2) infection. Little is known about the molecular interplay between viral infection and neoplastic transformation. The data regarding papillomavirus infections in human species show the inactivation of a number of tumour suppressor genes as basic mechanism of transformation. In this study the putative role of the tumour suppressor gene Fragile Histidine Triad (FHIT) in sarcoid tumour was investigated in different experimental models. The expression of the oncosuppressor protein was assessed in normal and sarcoid cells and tissue.
Nine paraffin embedded sarcoids and sarcoid derived cell lines were analysed for the expression of FHIT protein by immunohistochemistry, immunofluorescence techniques and western blotting. These analyses revealed the absence of signal in seven out of nine sarcoids. The two sarcoid derived cell lines too showed a reduced signal of the protein. To investigate the causes of the altered protein expression, the samples were analysed for the DNA methylation profile of the CpG island associated with the FHIT promoter. The analysis of the 32 CpGs encompassing the region of interest showed no significative differential methylation profile between pathological tissues and cell lines and their normal counterparts.
This study represent a further evidence of the role of a tumour suppressor gene in equine sarcoids and approaches the epigenetic regulation in this well known equine neoplasm. The data obtained in sarcoid tissues and sarcoid derived cell lines suggest that also in horse, as in humans, there is a possible involvement of the tumour suppressor FHIT gene in BPV induced tumours. DNA methylation seems not to be involved in the gene expression alteration. Further studies are needed to understand the basic molecular mechanisms involved in reduced FHIT expression.