Cadmium-treated(1.0 mg/L), Bt-treated(Bacillus thuringiensis protein, 0.1 mg/L), and Cd+Bt- treated flies were used as food for Pirata subpiraticus(P. subpiraticus), and the enrichment of Cd and Bt protein and expression of antioxidant enzymes in the spiders were investigated. In addition, RNA-seq was employed to analyze the differentially expressed genes(DEGs) of spiders under Cd+Bt treatment, and the enrichment and regulation pathways of DEGs was constructed using bioinformatics methods. It is found that compared with Cd(Cd content in the spiders was(6.965 ± 0.21 mg/kg) and Bt(Bt content in the spiders was(3.82 ± 0.28) μg/kg groups, the spiders in the Cd+Bt group showed higher Cd(12.23 ± 0.86) mg/kg and Bt(7.98 ± 0.49) μg/kg contents(P<0.05). In addition, compared with control, Cd and Bt groups, the Cd+Bt treatment increased the activities of glutathione peroxidase, glutathione S-transferase, and superoxide dismutase in spiders(P<0.05). RNA-seq was conducted on spiders in Cd+Bt treatment and in the control, a total of 6914 DEGs were obtained, and it is found that genes(519) involved in endopeptidase activity, hormone biosynthesis, and pheromone metabolism were all down-regulated obviously. Fourteen genes involved in oocyte meiosis were significantly down-regulated under Cd+Bt treatment. Through the analysis of transcription factors and protein interactions, we also found that transcription factors such as ZBTB participate in stress-related regulatory networks, which mainly affect the transcription and translation process of genetic information.