Amniotic membrane conditioned medium promotes cell death and inhibits proliferation of hepatocarcinoma HepG2 cells

RODRIGO RIEDEL; MARIANA JAIME; CECILIA VARONE; ANTONIO PÉREZ-PÉREZ; ORNELLA PAROLINI; JULIETA MAYMÓ; BERNARDO MASKIN; VICTOR SÁNCHEZ MARGALET

Mar del Plata

Congreso; ? LX Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2016

SAIC

The placenta and fetal membranes have recently been proposed as an important stem cells source for regenerative medicine. Stem cells derived from amniotic membrane offer considerable advantages over other stem cells because of the ease of collection, their low immunogenicity and inability to form tumors after transplantation. Moreover, minimal ethical and legal barriers are associated with their use. Epithelial amniotic cells isolated from the amnion express embryonic stem cells markers and have the ability to differentiate towards all three germ layers. Not only are amnion-derived stem cells applicable in regenerative medicine, but also have antitumoral properties. Hepatic failure is one of the major causes of morbidity and mortality and despite the development in therapies, hepatocarcinoma rates are high worldwide. A few studies have demonstrated the antittumoral effects of the amniotic membrane and their cells but little is known about the molecular and cellular mechanisms involved. The aim of this work was to study some aspects of cell death and proliferation induced by amniotic membrane conditioned medium (AM-CM) in hepatocarcinoma cells. We have analyzed the expression of proapoptotic proteins (p53, Caspase-3, PARP-1) by Western blot, in HepG2 cells treated with AM-CM. We have also analyzed cell proliferation by tritiated-thymidine (H3-T) incorporation assay and cell viability by MTT assay. We found a significant increment in p53 expression, Caspase-3 fragmentation and cleaved PARP-1 -measured by western blot-, after 24, 48 and 72 h of treatment with AM-CM in hepatocarcinoma cells. We have also observed that AM-CM significant decreased cell proliferation and viability, up to 7-fold compared with control, measured by H3-T incorporation and MTT assay, respectively. Finally, we have observed by immunofluorescence a diminished Ki-67 expression in HepG2 cells treated with AM-CM. Our results begin positioning amnion-derived stem cells as emerging candidates in anticancer therapy.