Stemness reducible effects of glyasperin A against NCCIT teratocarcinomas.

There is a great deal of interest in cancer stem cells (CSCs) in cancer therapy related to hallmarks of stemness and capacity for tumor relapse. Inhibition of CSCs by natural compounds represents a potential therapeutic approach. Thus, this study aimed to investigate potential effects of glyasperin A (GlyA) against CSCs by regulating possible stemness markers and signaling pathways as well. The NCCIT cell, a cell line derived from extragonadal germ cell tumors, was used as the cancer stem cell model for this study. The growth of teratocarcinoma cells under GlyA treatment in two-dimensional (2D) and three-dimensional (3D) cell culture patterns was revealed by MTT assay. The flow cytometry analysis was accessed to determine the expression of stemness surface markers and cell cycle arrest. In addition, the immunoblotting and human phospho-kinase array were applied to determine the possible target molecules in NCCIT cells under GlyA intervention. The collected results revealed that GlyA strongly inhibited the growth of NCCIT cells. The compound made cell cycle progression from the G0/G1 to the S phase. GlyA induced apoptosis by upregulating Bax and phosphorylated ERK1/2 protein levels. On the other hand, it also downregulated the expression of several transcription factors (Nanog, Oct4, c-Myc), which are closely correlated with stemness. Moreover, the levels of some proteins involved in the Akt/mTOR/IKK signaling pathways, which play critical roles in maintaining the self-renewal and proliferative features of CSCs, were also decreased. These results show that GlyA affects multiple targets that sustain the stemness features and proliferation or cell death of CSCs.