We aimed to delineate how pyrrolidine dithiocarbamate (PDTC) affects nuclear factor κB (NF-κB) and to determine its antitumor activity alone and in combination with cisplatin in human cervical cancer SiHa cells. The SiHa cells were treated with various concentrations of PDTC and/or cisplatin at various time intervals. Cell proliferation and apoptosis were determined using a water-soluble tetrazolium salt 8 assay and flow cytometry. Electrophoretic mobility shift assay was used to assess NF-κB activity. Pyrrolidine dithiocarbamate (2.5-100 µmol/L) was found to inhibit the growth of SiHa cell lines. Cisplatin (0.01-20.0 μg/mL) and PDTC (2.5-20.0 µmol/L) combined demonstrated additive inhibitive effects on cell growth and increased the level of apoptosis. In addition, PDTC blocked cisplatin-induced activation of NF-κB, leading to enhanced apoptosis and increased chemosensitivity to cisplatin. Taken together, PDTC has significant potential as a chemotherapy agent, alone or in combination with cisplatin.