Universidad de Buenos Aires. CONICET. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN). Ciudad Universitaria Pabellón 2, 4° piso, (1428), Buenos Aires, Argentina.
Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Ciudad Universitaria Pabellón 2, 4° piso, (1428), Buenos Aires, Argentina.
Sci Rep. 2019 Oct 2;9(1):14193. doi: 10.1038/s41598-019-50648-5.
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 antiproliferative, antiangiogenic and proapoptotic properties of the amniotic membrane. 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. In this work we aimed to study the antiproliferative properties of the human amniotic membrane conditioned medium (AM-CM) in hepatocarcinoma cells. In addition, we have analyzed the regulation of pro and antiOncomiRs expression involved in hepatocarcinoma physiology. We have determined by H-thymidine incorporation assay that AM-CM inhibits DNA synthesis in HepG2 cells after 72 h of treatment. AM-CM pure or diluted at 50% and 25% also diminished HepG2 and HuH-7 cells viability and cell number. Furthermore, AM-CM induced cell cycle arrest in G2/M. When proliferation mechanisms were analyzed we found that AM-CM reduced the expression of both Cyclin D1 mRNA and protein. Nuclear expression of Ki-67 was also reduced. We observed that this CM was able to promote the expression of p53 and p21 mRNA and proteins, leading to cell growth arrest. Moreover, AM-CM induced an increase in nuclear p21 localization, observed by immunofluorescence. As p53 levels were increased, Mdm-2 expression was downregulated. Interestingly, HepG2 and HuH-7 cells treatment with AM-CM during 24 and 72 h produced an upregulation of antiOncomiRs 15a and 210, and a downregulation of proOncomiRs 206 and 145. We provide new evidence about the promising novel applications of human amniotic membrane in liver cancer.
胎盘干细胞因其在治疗包括癌症在内的多种疾病方面的治疗潜力而成为关注焦点。有大量证据表明羊膜具有抗增殖、抗血管生成和促凋亡的特性。肝癌是世界上第五大癌症，预后和生存率较差。人们一直在寻找放射或化学疗法的替代方法。在这项工作中，我们旨在研究人羊膜条件培养基（AM-CM）对肝癌细胞的增殖抑制特性。此外，我们还分析了与肝癌生理学相关的原癌 miRNA 和抑癌 miRNA 表达的调节。通过 H-胸苷掺入测定法，我们确定 AM-CM 在处理 72 小时后抑制 HepG2 细胞的 DNA 合成。AM-CM 纯品或稀释至 50%和 25%也减少了 HepG2 和 HuH-7 细胞的活力和细胞数量。此外，AM-CM 诱导细胞周期停滞在 G2/M 期。当分析增殖机制时，我们发现 AM-CM 降低了 Cyclin D1 mRNA 和蛋白的表达。Ki-67 的核表达也减少了。我们观察到这种 CM 能够促进 p53 和 p21 mRNA 和蛋白的表达，导致细胞生长停滞。此外，AM-CM 诱导核内 p21 定位增加，通过免疫荧光观察到。随着 p53 水平的升高，Mdm-2 的表达下调。有趣的是，HepG2 和 HuH-7 细胞在处理 AM-CM 24 和 72 小时后，抗原癌 miRNA 15a 和 210 的表达上调，而原癌 miRNA 206 和 145 的表达下调。我们提供了关于人羊膜在肝癌中具有有前途的新应用的新证据。
