A versatile and sensitive quantum dot (QD)-based "signal-off" electrochemiluminescence (ECL) sensing system was constructed using target-initiated dual Mg2+-dependent DNAzyme (MNAzyme) recycling and catalytic hairpin assembly (CHA) amplification strategies. After the cascade amplification, numerous ferrocene-labeled Y-shaped DNA complexes generated on the QD-modified electrode surface. In the presence of hemin, moreover, the terminal sequence of the formed complex could assemble into hemin/G-quadruplex. Therefore, the highly efficient ECL quenching was achieved due to the multiple quenching mechanisms, including electron/energy transfer between ferrocene and QDs, the steric hindrance effects, and the horseradish peroxidase-mimicking activity of hemin/G-quadruplex. Furthermore, owing to the flexibility in regulating the recognition sequences of MNAzyme, the assaying targets can be programmed. Based on the cascade amplification and multiple ECL quenching mechanisms, the developed programmable "signal-off" ECL sensing platform demonstrates excellent sensitivity and the detection limits of 35.00 aM, 3.71 fM, and 0.28 pM (S/N = 3) for target DNA, aptamer substrate (ATP as a model), and ion (Ag+ as a model), respectively.