Preliminary Study On The Effects Of Differential Genes And Vitamin A On The Causes Of Hirschsprung’s Disease

SECTION ONE Differential expression of FOXA1, DUSP6, and HA117 in colon segments of Hirschsprung’s diseaseObjective To describe the expression profiles of FOXA1, DUSP6, and HA117 in different portions of the colon of patients diagnosed with Hirschsprung’s disease (HD).Methods Colon specimens were collected from34 HD patients and grouped into 3 segments:anastomosis, dilated segment and stenotic segment. Levels of FOXA1, DUSP6, and HA117 RNA were evaluated by real-time PCR. Levels of FOXA1 and DUSP6 protein were analyzed by immunohistochemistry and Western blottingResults The levels of FOXA1 and DUSP6 RNA were significantly lower in the stenotic segment compared to anastomosis (P< 0.05). The level of HA117 RNA was significantly higher in the stenotic segment compared to anastomosis (P< 0.05). In anastomosis, FOXA1 and DUSP6 were both expressed at the protein level in ganglion cells and nerve fibers between the circular and longitudinal muscles. In the stenotic segments, positive staining for FOXA1 and DUSP6 was diminished. The levels of FOXA1 and DUSP6 protein were significantly lower in the stenotic segment compared to anastomosis (P< 0.05).Conclusion Suppression of the FOXA1/DUSP6 signaling pathway may contribute to the development of HD. LncRNA HA117 may have an anti-differentiation function, and play a pivotal role in the progression of HD.SECTION TWO Expression of Vitamin A in Peripheral blood and Differential expression of RARa and HA117 in colon segments of children with Hirschsprung’s diseaseObjective To describe the expression profiles of Vitamin A in blood serum and RARa, HA117 in different portions of the colon of patients diagnosed with Hirschsprung’s disease (HD).Methods Peripheral blood and colon specimens were collected from 17 HD patients. The concentration of Vitamin A was detected in blood serum, and the colon specimens were grouped into 3 segments: anastomosis, dilated segment and stenotic segment. Levels of RARa mRNA and HA117 RNA were evaluated by real-time PCR. RARa protein expression were analyzed by Western Blot and immunohistochemistry.Results The concentration of Vitamin A in peripheral blood is 0.73±0.05μmol/L, which is below the normal level. The levels of RARα mRNA expression was significantly lower in the stenotic segment compared to anastomosis (P< 0.05). The level of HA117 RNA expression was significantly higher in the stenotic segment compared to anastomosis (P< 0.05). The levels of RARα protein expression was significantly lower in the dilated segment (P<0.05) and stenotic segment (P<0.05) compared to anastomosis.Conclusion The Vitamin A and RARα may contribute to the development of enteric nervous system (ENS). The suppression of the Vitamin A and RARα may result in HD.LncRNA HA117 may have an anti-differentiation function, and play a pivotal role in the progression of HD.