Rapid and precise detection of tumor biomarkers in serum is of great significance for early diagnosis, treatment assessment, and recurrence monitoring of cancers. The fabrication of user-friendly electrochemiluminescence (ECL) aptasensors with effective signal amplification is highly desirable. In this study, an ECL aptasensor was developed with luminol-based ternary ECL platform using dissolved oxygen as endogenous co-reactant and nanochannel-confined platinum nanoparticles (PtNPs) as co-reactant promoter, which can realize highly sensitive detection of the tumor biomarker, carcinoembryonic antigen (CEA). On the cost-effective indium tin oxide (ITO) electrode, amino-modified vertically-ordered mesoporous silica film (NH2-VMSF) was easily grown to form high-density nanochannel array with ultrasmall nanochannels. PtNPs were then in-situ synthesized within the nanochannels via electrodeposition. Ternary ECL system was successfully fabricated as confined PtNPs can catalyze the electroreduction of dissolved oxygen at negative potentials as well as electrochemical oxidation of luminol, enhancing the ECL signal of luminol. The recognitive aptamer (Apt) was covalently immobilized on the outer surface of NH2-VMSF, resulting in the fabrication of an aptasensor. In presence of CEA, a decreased ECL signal was obtained as the formed complex on the recognition interface hindered the diffusion of ECL emitter to the supporting electrode. Based on this signal turn-off mode, sensitive detection of CEA was achieved ranged from 10 fg mL−1 to 100 ng mL−1, with a low detection limit (DL, 0.4 fg mL−1). The aptasensor displayed high selectivity and good reproducibility. The nanochannel-based aptasensor and ternary ECL system demonstrates great potential in convenient and sensitive detection of cancer biomarker.