High levels of tumor-associated calcium signal transduction protein (TROP)-2 have been demonstrated to be strongly associated with tumor necrosis factor (TNF)-alpha levels in colon cancer. In the present study, the effect of TNF-alpha on the regulation of TROP-2 expression and its effect in colon cancer cell migration and invasion were investigated in vitro. TROP-2 protein levels were evaluated in HCT-116 human colon cancer cells cultured with various concentrations of TNF-alpha using western blot analysis. Changes in the migratory and invasive potential of the cells were evaluated using a wound healing and transwell assay, respectively. Then, TROP-2 expression was downregulated in cells by use of siRNA, and TROP-2 knockdown was confirmed at the mRNA and protein level by reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. The migration and invasion potential of cells transfected with TROP-2 siRNA was also evaluated. Levels of several mitogen-activated protein kinase proteins, namely p38, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), were detected in cells treated with TNF-a using western blot analysis. The results demonstrated that TROP-2 protein levels increased in cells treated with lower concentrations of TNF-alpha, but decreased in cells treated with higher concentrations of TNF-a, compared with untreated control. Maximum TROP-2 levels were observed in cells treated with 20 mu g/l TNF-alpha. Migration and invasion were enhanced in cells treated with 20 mu g/l TNF-alpha. When TROP-2 was silenced in colon cancer cells by siRNA, migration and invasion were significantly decreased compared with control cells. TNF-alpha stimulation activated the ERK1/2 pathway, but did not significantly affect p38 and JNK phosphorylation levels. Treatment with a specific ERK1/2 inhibitor suppressed the TNF-alpha-induced upregulation of TROP-2 and the TNF-alpha-induced colon cancer cell migration and invasion. In conclusion, the present results demonstrated that low concentrations of TNF-alpha significantly enhanced colon cancer cell migration and invasion by upregulating TROP-2 via the ERK1/2 signaling pathway.