Triclosan (TCS) is released into the terrestrial environment via the application of sewage sludge and reclaimed water to agricultural land. More attention has been paid to its effect on non-target soil organisms. In the present study, chronic toxic effects of TCS on earthworms at a wide range of concentrations were investigated. The reproduction, DNA damage, and expression levels of heat shock protein (Hsp70) gene of earthworms were studied as toxicity endpoints. The results showed that the reproduction of earthworms were significantly reduced (p < 0.05) after exposure to the concentrations ranges from 50 to 300 mg kg−1, with a half-maximal effective concentration (EC50) of 142.11 mg kg−1. DNA damage, detected by the comet assay, was observed and there was a clear significant (R2 = 0.941) relationship between TCS concentrations and DNA damage, with the EC50 value of 8.85 mg kg−1. The expression levels of Hsp70 gene of earthworms were found to be up-regulated under the experimental conditions. The expression level of hsp70 gene increased, up to about 2.28 folds that in the control at 50 mg kg−1. The EC50 value based on the Hsp70 biomarker was 1.79 mg kg−1. Thus, among the three toxicity endpoints, the Hsp70 gene was more sensitive to TCS in soil.