In view of the urgent need of determining polychlorinated biphenyls in the environment, we developed a highly sensitive and selective photoelectrochemical (PEC) aptasensor for determination of 3,3′,4,4′-tetrachlorobiphenyl (PCB77) by immobilizing aptamer on N-doped TiO2 nanotubes (N-doped TiO2 NTs). To improve analytical performance of the PEC sensor, the complementary DNA functionalized CdS quantum dots (DNA-CdS QDs) were introduced onto N-doped TiO2 NTs by hybridization. In addition of PCB77, owing to high affinity of aptamer to PCB77, PCB77-aptamer complexes were formed by being bound of PCB77 whilst DNA-CdS QDs were released from the sensing surface. The complexes with poor conductivity hindered the interfacial electron transfer, leading to the photocurrent decrease. The more important is the release of DNA-CdS QDs enhanced the photocurrent decrease, playing the role of signal amplification. The photocurrent change was utilized to detect PCB77 quantitatively. The PEC aptasensor exhibited excellent analytical performance for detection of PCB77 with wide linear range of 0.1–100 ng/L and a low detection limit of 0.1 ng/L. It manifested outstanding selectivity for PCB77 in control experiments by employing six interferents which had similar structure or coexisted with PCB77. Besides, the PEC aptasensor was used to detect the content of PBC77 in the environment.