Amniotic membrane conditioned medium inhibits tumoral cells proliferation and modulates related microRNAs expression in hepatocarcinoma models in vitro

RODRIGO RIEDEL; MARIANA JAIME; JULIETA MAYMÓ; BERNARDO MASKIN; CECILIA VARONE; ANTONIO PEREZ PEREZ; VICTOR SANCHEZ MARGALET

Mar del Plata

Congreso; LXIII Reunión anual SAIC SAI SAFIS 2018; 2018

The 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. We have previously demonstrated that the amniotic membrane conditioned medium (AM-CM) inhibits DNA synthesis and viability in hepatocarcinoma cells. In the present work we aimed to evaluate the antiproliferative properties of the AM-CM in hepatocarcinoma cells. We have analyzed the expression of key cell cycle proteins (cyclin D1, p53, p21, MDM-2) by Western blot and qRT-PCR, in HepG2 cells treated with AM-CM. In addition, we have analyzed the regulation of miR-15a, miR-210, miR-206 and miR-145 expression (pro and antiOncomiRs involved in hepatocarcinoma physiology) by qRT-PCR. We found that AM-CM reduced (5 ± 0.5 fold) the expression of both Cyclin D1 mRNA and protein. We observed that this conditioned medium was able to promote the expression (2.2 ± 0.3 fold) of p53 and (4 ± 0.05 fold) p21 mRNA and proteins, leading cells to growth arrest. Moreover, AM-CM induced an increase (3 ± 0.3 fold) in nuclear p21 expression comparing with the cytoplasmic one, observed by immunofluorescence assay. As p53 levels were increased, MDM-2 expression was downregulated (3.1 ± 0.2 fold), as expected. Interestingly, HepG2 and Huh-7 treatment with AM-CM produced an upregulation of antiOncomiRs 15a and 210, and a downregulation of the proOncomiRs 206 and 145. We provide new evidence about the promising novel applications of the human amniotic membrane in liver cancer.