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All trans retinoic acid (ATRA) plays an important role in many cellular processes and is a potent promising substance for cancer therapy. The self-renewal is a prominent feature of cancer stem cells that is tightly controlled by a number of specific genes, and is also mediated by the cell signaling pathways. The Notch signal pathway has been shown to be one of the few major molecular signaling pathways of cancer stem cells, which regulates self-renewal and survival of cancer stem cells. In this study, we showed that ATRA reduced the expression of important genes involved in self-renewal of cells including Sox2, KLF4, DMNT1 and MYC as well as TBGUT markers such as CD24, MUC1 and CD90. Furthermore, we indicate that the ATRA-induced expression of self-renewal genes and cancer stem cell markers of gastric cancer stem cells can be mediated by the regulation of the Notch signaling pathway.