The downregulation of VPS9D1-AS1 expression regulates cell proliferation, invation, migration, mobility and cell cycle. (A) Interference efficiency of sh-VPS9D1-AS1-1,2,3 in ESCC cells was evaluated by qRT-PCR. (B) CCK-8 assays were performed that VPS9D1-AS1 knockdown significantly inhibited the proliferation of Eca109 and TE-13 cells. (C) Transwell assays exhibited that VPS9D1-AS1 knockdown significantly inhibited the invation, and migration of Eca109 and TE-13 cells. (D) Wound healing assays were performed that VPS9D1-AS1 knockdown significantly inhibited the mobility of Eca109 and TE-13 cells. (E) VPS9D1-AS1 knockdown resulted in cell cycle arrest in Eca109 and TE-13 cells. (F) VPS9D1-AS1 knockdown resulted in decreased expression of cell cycle critical proteins in Eca109 and TE-13 cells. (Magnification: 200×,*P<.05, **P<.01, ***P<.001, and ****P<.0001).