Exploring cysteine regulation in cancer cell survival with a highly specific “Lock and Key” fluorescent probe for cysteine

Abstract

To probe the regulatory roles of cysteine (Cys) in cancer cell survival, a highly selective and sensitive fluorescent Cys probe <b>SiR</b> was developed by employing a novel "lock and key" strategy, which allows Cys to be detected without any interference or probe consumption caused by the intracellular high concentration of glutathione (GSH). Using <b>SiR</b>, we confirmed that inhibiting cystine (Cys₂) transporter system x_c ^- to deplete intracellular Cys is more efficient than inhibiting glutamate-cysteine ligase GCL to deplete intracellular GSH for sensitizing cancer cells to chemotherapy. Moreover, with the probe, a possible self-protection mechanism of cancer cells was indicated: when extracellular Cys sources are blocked, cancer cells could still survive by multidrug resistance protein transporter (Mrp1)-mediated export of intracellular GSH/GSSG as sources to supply intracellular Cys for resisting detrimental oxidative stress. Based on this finding, we further confirmed that abrogating the self-protection mechanism is an even more efficient strategy for sensitizing cancer cells to chemotherapy.