We found that capsaicin suppressed 5-FU-induced autophagy via activation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway in CCA cells, which rendered the cells more susceptible to 5-FU both in vitro and in vivo. Here, AKT1 is linked to cholangiocarcinoma.