ESPU Congress 2017 - Abstract book

CONGRESS OF THE ESPU

15:38–15:41

S2-2 (PP)

AUTOPHAGY, APOPTOSIS AND CELL PROLIFERATION

IN EXTROPHY-EPISPADIAS COMPLEX

Mahsa SHABANINIA, Ali TOURCHI, Heather DI CARLO and John GEARHART

The James Buchanan Brady Urological Institute, The Johns Hopkins School of Medicine, Department of Urology,

Baltimore, USA

PURPOSE

Very few pathophysiological mechanisms have been proposed as the etiology of bladder exs-

trophy (BE). Autophagy, or type II programmed cell death pathway, is an evolutionary conserved

process involving intracellular degradation and recycling of cytoplasmic organelles. A basal level

of autophagy is detected in most tissues, maintaining cellular homeostasis and viability through

development and differentiation of eukaryotic organisms. Herein, the authors investigated the state

of autophagy and its interactions with cells apoptosis and proliferation in patients with BE.

MATERIAL AND METHODS

Primary cultures of bladder smooth muscle cells were established from patients with successful neo-

natal bladder closure (group 1, N=5), delayed closure due to small bladder template (group 2, N=5)

and vesicoureteral reflux as control (group3, N=5). The myogenicity of the cultures was determined

using anti-desmin antibody. Immunofluorescence staining for LC3 was used to detect autophagy.

Cells apoptosis was assessed using TUNEL assay, 4′, 6-diamidino-2-phenylindole staining. Cellular

proliferation was assessed by image analysis of immunofluorescence staining for Ki-67.

RESULTS

Immunohistochemical staining revealed consistent positivity (greater than 95%) for Desmin in all

cultures that confirms the myogenicity of them. Apoptosis was significantly higher in delayed closure

group compared to other groups. Autophagy marker (LC3) was more expressed in delayed closure

group compared to the other groups. Cellular proliferation was significantly lower in delayed closure

group compared with control and successful neonatal closure groups.

CONCLUSIONS

Our results confirms that there are distinct differences in bladder smooth muscle cell function be-

tween control, successful neonatal closure and delayed closure cases due to small bladder template

which persist in culture. Children with slower bladder growth and small bladder templates showed

up-regulated autophagic process and increased apoptotic indices while experiencing a dramatic

decrease in their bladder smooth muscle cells proliferation.