Peer Review #2 of "Discovery of potential targets of Triptolide through inverse docking in ovarian cancer cells (v0.1)"

Abstract

Triptolide (TPL) is proposed as an effective anticancer agent known for its anti-proliferation of a variety of cancer cells including ovarian cancer cells. Although some studies have been conducted, the mechanism by which TPL acts on ovarian cancer remains to be clearly described. Herein, systematic work based on bioinformatics was carried out to discover the potential targets of TPL in SKOV-3 cells. TPL induces the early apoptosis of SKOV-3 cells in a dose-and time-dependent manner with an IC 50 =40±0.89 nM when cells are incubated for 48h. Moreover, 20 nM TPL significantly promotes early apoptosis at a rate of 40.73%. Using a self-designed inverse molecular docking protocol, we fish the top nineteen probable targets of TPL from the target library which are built on 2250 proteins extracted from the Protein Data Bank. The 2D-DIGE assay reveals that the expression of eight genes is affected by TPL. The results of western blotting and qRT-PCR assay suggest that 40 nM of TPL up-regulates the level of Annexin A 5 (6.34±0.07 fold) and ATP syn thase (4.08±0.08 fold), and down-regulates the level of β-Tubulin (0.11±0.12 fold) and HSP90 (0.21±0.09 fold). More details of TPL affecting on Annexin A5 signaling pathway will be discovered in the future. Our results define some potential targets of TPL, with the hope that this agent could be used as therapy for the preclinical treatment of ovarian cancer.