Nano-drug delivery systems (nanoDDS) have been extensively investigated clinically to improve the therapeutic effect of anticancer drugs. However, the complicated synthesis during the preparation as well as the potential drug leakage during transportation has greatly limited their general application. In this work, a calixarene-integrated nanoDDS (CanD) that achieves tumor-targeted delivery and tracking of anti-cancer drugs in vivo is presented. The hypoxia-responsive calixarene (SAC4A) exhibits high binding affinity to a series of anti-cancer drugs and rhodamine B (RhB) under normoxic condition while decreasing the binding affinity under hypoxic condition, which leads to the drug release and fluorescence recovery simultaneously. Furthermore, the hypoxia-responsiveness of SAC4A conveys CanD with tumor-targeting ability, resulting in the enrichment of the drug in tumors and enhancement in tumor suppression in mice. Moreover, CanD could become a general platform allowing the delivery of a wide scope of anti-cancer drugs that have strong host-guest interaction with SAC4A.