Possible Mechanism Research Of Autophagy Induced By Folate Deficiency Via MiRNAs

Folic acid is a form of water-soluble vitamin B9, as a carrier of one carbon unit, which mainly involved in two biochemical process of DNA synthesis and DNA methylation. Folate deficiency can result in many diseases such as birth defects, aging and tumor. It can also induce cell death. Autophagy is a type of cell death which is a basic catabolic mechanism of survival for cells with fasting. The previous study of our team has confirmed that apoptosis can be induced by folate deficiency in mouse embryonic stem cells and miRNAs are the key factor of proliferation and apoptosis in embryo development. Other study have demonstrated that autophagy play a key role in development of vertebrates. However, the relationship between folic acid, miRNA and autophagy has not been characterized. In this study, mouse embryonic stem cells (mESCs) were used for research. The aim is to exploring whether folate deficiency could induce autophagy and whether miRNAs could involve in this process. Then we tested critical proteins of autophagy, screened and verified the miRNAs and mRNAs associated with autophagy so as to elaborate the relationship between folate deficiency, miRNA regulation and autophagy with multi-dimension.ObjectiveTo study the relationship between folate deficiency, autophagy and miRNAs, and the possible regulating mechanism in mESCs.Methods1. mESCs were treated with both normal folate levels (Control) and low folate levels (folate deficiency, FD medium)for36hours. Then CCK-8assay was applied to assessing the proliferation of mESCs in these two cultured systems. After cell harvesting, autophagy-related molecules such as LC3B and Beclinl were tested by Western Blotting and real-time PCR.