RNA-seq in liver cancer cell and cinobufagin treatment cells

Cinobufagin, belonged to a kind of cardiotonic steroids (CSs), derived from the extracts of toad venom, which presented the significant anticancer properties as inhibitors of Na+/K+-ATPase (NKA) in many cancer cells. Nowadays, cinobufagin was usually used to apply for clinical treatments of patients in advanced stage of cancer, and improved their quality of life and prolonged their survival period. Unfortunately, long-term drug treatment had also led to multi-drug-resistance (MDR) likely other chemotherapy drugs. However, this detailed mechanism was not still clear. In the present study, we noticed that cinobufagin could trigger the protective autophagy to suppress cells apoptosis in liver cancer HepG2 and Huh-7 cells by inhibition of PI3K-AKT-mTOR pathway using RNA-seq method. We also confirmed that cinobufagin could inhibit cells proliferation and induce cells apoptosis, and generated cells autophagy by up-regulation expression of LC3B-II, Beclin 1 and Atg12-atg5. More importantly, autophagy inhibitor (MRT68921) enhances the anti-proliferation and pro-apoptosis effects of cinobufagin in HCC cells. Therefore, we thought that a combination of cinobufagin and autophagy inhibitors may be a potential effective strategy in treatment of HCC. Overall design: Examination of cinobufagin treatment for 0, 12, and 24 h in 2 cell types.