In vitro studies of the TGFbeta/Smad pathway induced by boron neutron capture therapy (BNCT)

PERALTA T; ROSSICH L; NIEVAS S; CARPANO M; CUROTTO P; THORP S; POZZI E; JUVENAL G; DAGROSA MA

Mar del Plata

Congreso; LXIV Reunion anual de la sociedad argentina de investigacion clinica; 2019

Sociedad Argentina de Investigacion Clinica

Introduction: BNCT is based on the nuclear reaction 10B (n, α)7Li and the radiation field produced is a mixture of high and low LET components which activates the DNA damage response (DDR). It is well known the role of TGF beta1 in homoeostatic growth control; however it has been less studied its complex role in regulating responses to genotoxic stress (through transcription factors Smads). The aim of these studies was to analyze the TGF beta/Smad signaling pathway as part of the DDR arising from BNCT. Materials and Methods: HT29 human colon cancer cells were seeded in bottles of 25cm2 and distributed in 4 groups: 1) Control; 2) BNCT (BPA + neutrons); 3) NCT (neutrons alone) and 4) Bystander effect from BNCT. The irradiation was carried out in RA3 reactor (Neutron flux of 1.1010 n / cm2sec). After 2 h of incubation at 37 °C, the total RNA was extracted with Trizol and real time PCR was performed for each gene: TGF beta1, Smad2 Smad7, ATM and ATR. Results: the expression of TGF beta1 and Smad7 increased in all the BNCT and NCT groups respect to the Control (p˂0.05). The indirectly irradiated group (Bystander) showed an increase only for TGF beta 1. The expression of Smad2 decreased respect to the Control group in all the irradiated groups. ATM showed a significative decrease, while ATR showed an increase (p