“UNRAVELLING THE MECHANISMS OF HUMAN AMNIOTIC MEMBRANE PROAPOPTOTIC EFFECT IN CELULLULAR HEPATOCARCINOMA MODELS”

LUCIANO PÉREZ; RODRIGO RIEDEL; NATALY DE DIOS; ANTONIO PÉREZ-PÉREZ; MARIANA JAIME; VICTOR SÁNCHEZ MARGALET; CECILIA LAURA VARONE; JULIETA MAYMÓ

MAR DEL PLATA

Congreso; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2022

Sociedad Argentina de Investigación Clínica

Unravelling the mechanisms of human amniotic membrane proapoptotic effect in cellular hepatocarcinoma modelsAuthors: Luciano A. Pérez1, Rodrigo Riedel1, Nataly De Dios1, Antonio Pérez-Pérez2, Mariana Jaime3, Víctor Sánchez-Margalet3, Cecilia L. Varone1 and Julieta Maymó1.Affiliations:1Biological Chemistry, IQUIBICEN, CONICET-FCEN, UBA, Ciudad Autónoma de Buenos Aires, Argentina2Medical Biochemistry and Molecular Biology, Sevilla University, Sevilla, Spain3Maternity, Posadas Hospital, Buenos Aires, ArgentinaThe stem cells, and particularly the placental stem cells, have called the focus of attention for their therapeutic potential to treat different diseases, including cancer. There is plenty evidence about the anti-tumoral effects of the human amniotic membrane given by their antiproliferative, antiangiogenic and proapoptotic properties. The amnion and its cells both secrete unknown factors and physically interact with tumor cells. Liver cancer is the fifth cause of cancer in the world, with a poor prognosis and survival. Alternative treatments to radio- or chemotherapy have been searched. A few studies have demonstrated the antitumoral effects of the amniotic membrane and their stem cells, but little is known about the molecular and cellular mechanisms involved We have previously demonstrated that the amniotic membrane conditioned medium (AM-CM) inhibits hepatocarcinoma cells proliferation and survival. The aim of this work was to study the apoptotic mechanisms involved in hepatocarcinoma cells death after treating them with AM-CM. First, we have observed that AM-CM induced both early and late apoptosis process, in HepG2 and Huh-7 hepatocarcinoma cells, evaluated by Annexin V/PI staining and flow cytometry analysis. In this way, we have also detected late apoptosis occurrence in HepG2 cells treated during 72 h with AM-CM, determined by DNA fragmentation assay. When analyzing the intrinsic pathway, we have observed that AM-CM treatment promoted an increase in Bax/Bcl-2 ratio and in cytochrome c expression, measured by Western blot and immunofluorescence. The extrinsic pathway was also analyzed. Thus, we determined by Western blot a decrease in total Caspase-8 and Bid expression, in HepG2 cells treated during 24 and 72 h with AM-CM. Our results position amnion-derived stem cells as emerging candidates in anticancer therapy.