Polychlorinated biphenyls (PCBs) are a group of chlorinated organic compounds. They are persistent in the environment and can threaten the health of humans and wildlife. Urban stormwater runoff is considered as an important source of PCBs to aquatic environments. The objective of this study is to provide information on the occurrence and removal of PCBs in stormwater; specifically, the occurrence, concentrations, and biological transformations of stormwater PCBs were studied together with their removal. Concentrations of 209 PCB congeners were determined in surface stormwater sediments collected from various roadway sites and bioretention media. The total PCB concentrations ranged from 8.3 to 57.4 ng/g dry weight (dw), with a mean value of 29.2 ng/g dw. Land use had an impact on the concentration of PCBs, where higher stormwater sediment PCB concentrations were found in dense urban areas (average: 39.8 ± 10.5 ng/g) compared to highways passing through greenspace (average: 18.0 ± 0.4 ng/g). PCB sorption tended to increase with the concentration of total organic carbon (TOC) and smaller particle size (< 75 µm) of stormwater particulate matter. In bioretention core samples, PCB concentrations decreased with bioretention media depth (from 30.0 ± 2.0 ng/g at the surface to 21.2 ± 4.8 ng/g at 40 cm depth), and with distance from the stormwater entrance (from 38.4 ± 2.3 ng/g at the entrance to 33.2 ± 2.9 ng/g at 3 m distance). A non-Aroclor congener, PCB 11, was detected in all samples, likely originating from yellow road paint. Putative organohalide respiring bacteria within Chloroflexi and aerobic PCB degrading bacteria containing the functional genes encoding for biphenyl dioxygenase (bphA) and ring cleavage (bphC) were detected in some of the stormwater sediments and bioretention media. The presence of such bacteria and a higher level of ortho-chlorinated biphenyls indicated the potential of PCB biotransformation in these samples. The performance of an on-campus bioretention indicated that bioretention is effective in removing PCBs from stormwater, with 64–92% reduction of dissolved PCB concentrations. Overall, urban stormwater is an important environmental source of PCBs. Bioretention has the potential to remove PCBs from stormwater via adsorption and biotransformation.