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Aberrant microRNA (miRNA) expression has been linked to cancer development. In this study, we aimed to investigate whether the anti-cancer effect of miRNA-299-3p on laryngeal cancer Hep-2 cells is mediated through targeting human telomerase reverse transcriptase (hTERT). The expression of miR-299-3p in laryngeal cancer Hep-2 cells and human osteosarcoma U2OS cells was quantified by stem-loop-mediated reverse transcription quantitative polymerase chain reaction. miR-299-3p mimic was transfected into Hep-2 cells to induce overexpression of miR-299-3p. A CCK-8 assay was performed to identify the effects of miR-299-3p overexpression on the proliferation of Hep-2 cells. Western blot analysis was carried out to determine the expression of hTERT protein. A significant decrease was noted in the expression of miR-299-3p in Hep-2 cells compared with that of U2OS cells (P<0.05). Overexpression of miR-299-3p resulted in a notable inhibition of cellular proliferation (P<0.05), as well as downregulation of hTERT mRNA and protein in Hep-2 cells (P>0.05). The expression of miR-299-3p is downregulated in human laryngeal cancer Hep-2 cells. Overexpression of miR-299-3p inhibits Hep-2 cell growth by targeting the 3'-untranslated region of hTERT mRNA. [ABSTRACT FROM AUTHOR]