Hydrogen Sulfide Mediates Tumor Cell Resistance to Thioredoxin Inhibitor

Thioredoxin (Trx) is a pro-oncogenic molecule that underlies tumor initiation, progression and chemo-resistance. PX-12, a Trx inhibitor, has been used to treat certain tumors. Currently, factors predicting tumor sensitivity to PX-12 are unclear. Given that hydrogen sulfide (H₂S), a gaseous bio-mediator, promotes Trx activity, we speculated that it might affect tumor response to PX-12. Here, we tested this possibility. Exposure of several different types of tumor cells to PX-12 caused cell death, which was reversely correlated with the levels of H₂S-synthesizing enzyme CSE and endogenous H₂S. Inhibition of CSE sensitized tumor cells to PX-12, whereas addition of exogenous H₂S elevated PX-12 resistance. Further experiments showed that H₂S abolished PX-12-mediated inhibition on Trx. Mechanistic analyses revealed that H₂S stimulated Trx activity. It promoted Trx from the oxidized to the reduced state. In addition, H₂S directly cleaved the disulfide bond in PX-12, causing PX-12 deactivation. Additional studies found that, besides Trx, PX-12 also interacted with the thiol residues of other proteins. Intriguingly, H₂S-mediated cell resistance to PX-12 could also be achieved through promotion of the thiol activity of these proteins. Addition of H₂S-modified protein into culture significantly enhanced cell resistance to PX-12, whereas blockade of extracellular sulfhydryl residues sensitized cells to PX-12. Collectively, our study revealed that H₂S mediated tumor cell resistance to PX-12 through multiple mechanisms involving induction of thiol activity in multiple proteins and direct inactivation of PX-12. H₂S could be used to predict tumor response to PX-12 and could be targeted to enhance the therapeutic efficacy of PX-12.