Chick Early Amniotic Fluid (ceAF) Deters Tumorigenesis via Cell Cycle Arrest and Apoptosis

SIMPLE SUMMARY: Amniotic fluids have been utilized extensively in several clinical medicine applications, including cutaneous wound healing, myocardial infarctions, and regeneration of organs, as they are the source of many key components, signaling molecules, cytokines, and peptides. Here in this study, we reported the applications of chick early amniotic fluid (ceAF) against tumorigenesis and showed how ceAF contributes to significant inhibition of metastasis of cancer cells, including breast cancer and colon carcinoma cell lines, and compared with a regular cell line. Our study also highlights the efficacy of ceAF in a dose-dependent manner in a different type of cancers. The in vivo study and biochemical parameters of the tumor xenografts also provided similar findings indicating the promising therapeutic potentials of ceAF. ABSTRACT: In recent years, amniotic fluids have gained attention in cancer research. They have an influential role in protecting embryos against several anomalies. Chick early amniotic fluid (ceAF)—amniotic fluid isolated from growing chicken—has been used in many other studies, including myocardial infarctions and skin regeneration. In this study, we employed ceAF’s promising therapeutic applications against tumorigenesis in both in vitro and in vivo studies. We selected three robust proliferating tumor cell lines: BCaP37, MCF7, and RKO. We found that selective dosage is required to obtain maximum impact to deter tumorigenesis. ceAF not only disrupted the uniform colonies of tumor cell lines via disturbing mitochondrial transmembrane potential, but also arrested many cells at growing G1 state via working agonistically with aphidicolin. The significant inhibition of tumor metastasis by ceAF was indicated by in vivo models. This leads to apoptosis analysis as verified by annexin-V staining stays and immunoblotting of critical proteins as cell cycle meditators and apoptosis regulators. Not only on the protein level, but we also tested ceAF’s therapeutic potentials on mRNA levels as indicated by quantitative real-time PCR summarizing the promising role of ceAF in deterring tumor progression. In conclusion, our study reveals the potent role of ceAF against tumorigenesis in breast cancer and colon carcinoma. Further studies will be required to determine the critical components present in ceAF and its purification to narrow down this study.