Expression levels and potential roles of DPF3 and its related genes of Smarcd3 and MEF2 in colonic tissues from patients with Hirschsprung's disease

Objective To detect the expression levels of DPF3, Smarcd3 and MEF2 in proximal anastomosis (control) and stenotic segments of colonic tissues from patients with HD and analyze their potential pathogenic roles in Hirschsprung's disease (HD). Methods All resected colonic tissues were obtained randomly from the HD patients at Children's Hospital of Chongqing Medical University between June 2012 and April 2014. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used for detecting DPF3, Smarcd3 and MEF2 mRNAs and protein expression in different portions of colonic tissues respectively. Immunohistochemistry and light microscope digital radiography were used for analyzing protein staining. GraphPad Prism 5 (GraphPad Software, Inc, USA) and SPSS statistical software (version 17.0) were used for spreadsheet and statistical analysis. Results The expression levels of DPF3 mRNA and protein were 1.76±1.30, 1.07±0.93 and 2.31±0.62, 1.95±0.57 in proximal anastomosis and stenotic segments respectively. The expression levels of Smarcd3 mRNA and protein were 0.68±0.62, 0.38±0.42 and 0.85±0.43, 0.61±0.34 in proximal anastomosis and stenotic segments respectively. And the expressions of MEF2 mRNA and protein in proximal anastomosis and stenotic segments were 1.01±0.63, 0.68±0.39 and 2.29±0.38, 1.83±0.76 respectively. Besides, both mRNA and protein of DPF3, Smarcd3 and MEF2 were remarkably less in stenotic segments than those in proximal anastomosis. And positive staining of DPF3, Smarcd3 and MEF2 was observed in proximal anatomosis. However, no significant staining was found in stenotic segment. Conclusions DPF3, Smarcd3 and MEF2 may be involved in the process of HD. Key words: Congenital megacolon; Polymerase chain reaction; Gene expression