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. 2015 Sep 15;8(9):14999-5004.
eCollection 2015.

Interferon-λ1 suppresses invasion and enhances autophagy in human osteosarcoma cell

Dongmei Gao  1 , Jun Zhao  2 , Xiuyi Li  1 , Yuanyuan Xia  1 , Yu Cai  1 , Jiwen Pan  1 , Hong Zhou  1 , Yong Fang  1 , Shouzhu Zhang  1 , He Wen  1
Affiliations

Interferon-λ1 suppresses invasion and enhances autophagy in human osteosarcoma cell

Dongmei Gao et al. Int J Clin Exp Med. .

Abstract

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 100 ng/mL Interferon-λ1 for 48 h. Osteosarcoma cell cells treated with 100 ng/mL Interferon-λ1 for 48 h developed autophagy some-like characteristics, including single or double-membrane vacuoles containing intact and degraded cellular debris.

Conclusions: Interferon-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.

Keywords: Interferon-λ1; acridine orange; autophagy; transmission electron microscopy.

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Figures

Figure 1
Figure 1
Cell invasion assay after treatment with IFN-λ1 for 48 h. A. Representative cell invasion assay for MG-63 cells. Invasion assays were carried out in 12-well transwell inserts of polycarbonate filters with 12.0 μm pores coated with 200 μl of 0.1% matrigel. After 48 h incubation period, the membranes were collected and stained. A significant reduction in the number of invaded cells indicated the decrease in invasive capacity. B. Quantitative evaluation of matrigel invasion assay. The data represented are mean ± SD of 10 randomly selected microscopic fields from 3 independent wells (*P < 0.01 compared with the control mean values). 1: Control group. 2: Treated with 1 ng/ml IFN-λ1 group. 3: Treated with 10 ng/ml IFN-λ1 group. 4: Treated with 100 ng/ml IFN-λ1 group.
Figure 2
Figure 2
Antiproliferative effect of IFN-λ1. MG-63 cells were treated with IFN-λ1. Cell viability was determined by MTT assay. 1: Control group. 2: Treated with 1 ng/ml IFN-λ1 group. 3: Treated with 10 ng/ml IFN-λ1 group. 4: Treated with 100 ng/ml IFN-λ1 group. (*P < 0.01 compared with the control mean values).
Figure 3
Figure 3
Modulation of autophagy by IFN-λ1 in MG-63 cells. Cells were treated with IFN-λ1 for 48 h at concentrations of 1 ng/ml, 10 ng/ml, and 100 ng/ml. Cells were then stained with acridine orange 1: Control group. 2: Cells treated with 1 ng/ml IFN-λ1. 3: Cells treated with 10 ng/ml IFN-λ1. 4: Cells treated with 100 ng/ml IFN-λ1.
Figure 4
Figure 4
Modulation of autophagy by IFN-λ1 in MG-63 cells. Cells were treated with IFN-λ1 for 48 h at concentrations of 1 ng/ml, 10 ng/ml, and 100 ng/ml. Cells were then stained with MDC. 1: Control group. 2: Cells treated with 1 ng/ml IFN-λ1. 3: Cells treated with 10 ng/ml IFN-λ1. 4: Cells treated with 100 ng/ml IFN-λ1.
Figure 5
Figure 5
Autophagic vacuoles quantified by flow cytometry. 1: Control group. 2: Cells treated with 1 ng/ml IFN-λ1. 3: Cells treated with 10 ng/ml IFN-λ1. 4: Cells treated with 100 ng/ml IFN-λ1. (#P < 0.05 relative to control values and *P < 0.01 relative to control values).
Figure 6
Figure 6
Transmission electron images of MG-63 cells treated with IFN-λ1. A: Control group. B: Cells treated with 100 ng/ml IFN-λ1 group.
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