A topsoil sample obtained from a highly industrialized area was tested on CALUX® cell line and exposed cells processed with proteomic and bioinformatics analyses. GC-MS/MS analysis of topsoil extracts was also performed to confirm PCDD, PCDF and dl-PCBs presence. CALUX® and GC-MS/MS data confirmed the presence of PCDD/Fs and dl-PCBs in the topsoil extract. Proteomic analysis of the cells exposed to the soil extracts identified 43 differential proteins. Enrichment analysis highlights biological processes such as cellular response to chemical stimulus, to stress, response to inorganic substance, regulation of translation, regulation of apoptotic process as well as response to organonitrogen compounds in light of particular drugs and compounds, extrapolated by bioinformatics and linked to the identified proteins modifications. These results confirm the complex epidemiological situation occurring among Taranto inhabitants and underline the need to further investigate the presence and sources of inferred chemicals in Taranto soils. The combined analysis reveals a more complex scenarios of chemicals able to affect cellular pathways and leading to toxicities rather than those identified by the bioassay. The approach to couple proteomic to CALUX® is a promising tool and deserves further investigation to be applied in soil pollution monitoring and risk assessment.