Autophagy and apoptosis are considered as two main kinds of programmed cell death. The crosstalk between autophagy and apoptosis is crucial for illustration of anti-tumor drug’s effects. Triptolide, a diterpene compound extracted from Tripterygium wilfordii Hook F, attracts increasing attention from researchers worldwide for its broad anti-tumor spectrum. Triptolide could bind to XPB (a subunit of the transcription factor TFⅡH) and leads to RNA polymerase Ⅱ-mediated transcription inhibition. Also, triptolide induces autophagy and apoptosis processes in cancer cells. Through regulating different kinds of related proteins, tumor repressor p53, plays key roles in the crosstalk between autophagy and apoptosis. Thus, in this study, the authors focused on triptolide induced p53-dependent autophagy and apoptosis processes in HeLa cells, and to demonstrate the regulation of crosstalk through p53-dependent transcription inhibitor and autophagy inhibitors treatment. Using CCK-8 assay and clonogenic assay, triptolide shows significant inhibition effect on HeLa cells. The apoptosis analysis by Western blot has shown that triptolide induces the cleavage of caspase3 and nuclear poly (ADP-ribose) polymerase (PARP). Immuno-fluorescence and Western blot assay have shown that triptolide also leads to LC3-Ⅱ accumulation and p62 degradations. Collectively, triptolide induces autophagy and apoptosis in a time- and dose-dependent manner. Meanwhile, p53 is significantly up-regulated and the mammalian target of rapamycin (mTOR) is down-regulated with triptolide treatment, and ultimately results in activation of autophagy and apoptosis. Western blot assay have shown that triptolide-induced autophagy and apoptosis are partly inhibited by utilizing p53-dependent transcription inhibitor (Pifithrin-α). Furthermore, the combination treatment of autophagy inhibitors with triptolide enhances triptolide induced apoptosis, and significantly improves inhibition effect of triptolide in HeLa cells. Triptolide induces p53-dependent autophagy and apoptosis in HeLa cells. p53 functions as a key regulator in the triptolide induced autophagy and apoptosis crosstalk.