Abstract: Objective：To explore the effects of hypoxia and thymic stromal lymphopoietin (TSLP) on the invasion of cervical cancer cells in vitro. Methods：Enzyme linked immunosorbent assay (ELISA) was performed to analyze the effects of normoxia and hypoxia on the secretion level of TSLP, and to analyze the effects of recombinant human TSLP (rhTSLP) protein on the expression levels of matrix metalloprotein-2 (MMP-2), MMP-9， and IL-8 in cervical cancer cell lines HeLa and CaSki. Matrigel invasion assay was used to investigate the effects of rhTSLP, hypoxia, and antihuman TSLP （aTSLP） neutralizing antibody on the invasiveness of HeLa and CaSki cells in vitro. Results：Under hypoxia exposure, the levels of TSLP secretion in HeLa and CaSki cells were significantly increased (P＜0.01). RhTSLP (100 ng/mL) stimulation in vitro significantly enhanced the invasiveness of HeLa and CaSki cells (P＜0.001), and promoted the productions of MMP-2, MMP-9， and IL-8 (P＜0.01). The invasiveness of HeLa and CaSki cells can be promoted significantly under hypoxia exposure (P＜0.001). In contrast, aTSLP (5 μg/mL) inhibited the invasiveness of HeLa and CaSki cells (P＜0.05), and partly inhibited the stimulatory effect of hypoxia on invasiveness of HeLa and CaSki cells (P＜0.05). Conclusions：Hypoxia facilitates the invasion of cervical cancer cells in vitro by up-regulating the secretion of TSLP.