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Long non-coding RNA (lncRNA) are key regulatory molecules that are implicated in diverse biological processes and human diseases, including preeclampsia. However, their expression and functions in the progression of preeclampsia remains largely unclear. In this study, lncRNA DLX6-AS1 was confirmed to be significantly upregulated in the placentas of patients with preeclampsia, compared with normal controls. Overexpression of DLX6-AS1 dramatically decreased proliferation, migration and invasion in trophoblast JEG3 and HTR-8/SVneo cells. Luciferase activity and RNA pull-down assays showed that DLX6-AS1 interacted with miR-376c, and that overexpression of DLX6-AS1 significantly reduced expression of miR-376c in HTR-8/SVneo cells. Also, miR-376c targeted and downregulated GADD45A in HTR-8/SVneo cells. Overexpression of GADD45A effectively attenuated a miR-376c-induced increase in the proliferation, migration and invasion of HTR-8/SVneo cells. Further research showed that DLX6-AS1-induced inhibition of trophoblast proliferation, migration and invasion was effectively neutralized by GADD45A knockdown and miR-376c overexpression. Taken together, these findings suggest that DLX6-AS1 may contribute to preeclampsia by impairing proliferative, migratory and invasive abilities of trophoblasts via the miR-376c/GADD45A axis.lncRNA DLX6-AS1 is upregulated in placentas from women with preeclampsia.DLX6-AS1 inhibits trophoblastic migration and invasion.DLX6-AS1 affects trophoblastic migration and invasion via miR-376c/GADD45A pathway.