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Objective: The purpose of the present study was to determine whether type Ⅲ IFN can modulate the autophagic response in human osteosarcoma cell.Methods: Human osteosarcoma cell were treated with Interferon-λ1.We investigated that Interferon-λ1 could inhibit the invasive ability of osteosarcoma cells by Matrigel invasion assay.Autophagy were assessed by acridine orange staining, MDC staining and Transmission electron microscopy.Results: In this study, we found that Interferon-λ1 could inhibit the invasive ability of osteosarcoma cells.Acridine orange staining and MDC staining showed that Interferon-λ1 triggered the accumulation of acidic vesicular and autolysosomes in osteosarcoma cell.The acridine orange osteosarcoma cell ratios were 3.6±0.5%, 4.5±0.8%, and 12.4±1.7% after treatment with 1, 10, and 100ng/mL Interferon-λ1 for 48 h.Osteosarcoma cell cells treated with 100 ng/mL Interferon-λ1 for 48 h developed autophagosome-like characteristics, including single-or double-membrane vacuoles containing intact and degraded cellular debris.Conclusions: Intefferon-λ1 could inhibit the invasive ability of osteosarcoma cells.Autophagy can be induced in a dose-dependent manner by treatment with Interferon-λ1 in osteosarcoma cell.