The present study was performed to investigate the expression of HOXA9 in human laryngeal squamous cell carcinoma and its possible roles in the progression. The levels of HOXA9 mRNA and protein were evaluated in human laryngeal squamous cell carcinoma. Hep-2 cells were transfected with h-HOXA9-siRNA. CCK-8 was used to analyze cell proliferation. Flow cytometry (FCM) was used to analyze cell cycle. The mobility of cells was tested by transwell migration assay. The expression of HOXA9 in laryngeal squamous cell carcinoma was significantly higher than normal mucosa tissues. In in vitro experiments, downregulation of HOXA9 strongly inhibited cell growth in Hep-2 by arresting cells in G1 phase (p < 0.05). Transwell migration assay showed that more HOXA9-negative cells migrated to the lower side of the membrane than positive ones (p < 0.01). HOXA9 acts as an oncogene in laryngeal squamous cell carcinoma. It could promote the proliferation and migration of Hep-2 cells.