Format

Send to

Choose Destination
Connect Tissue Res. 2018 Oct 2. doi: 10.1080/03008207.2018.1530770. [Epub ahead of print]

Reactive oxygen species are responsible for the cell aggregation and production of pro-inflammatory mediators in phorbol ester (PMA)-treated U937 cells on gelatin-coated dishes through upregulation of autophagy.

Author information

1
a China-Japan Research Institute of Medical and Pharmaceutical Sciences , Shenyang Pharmaceutical University , Shenyang 110016 , China.
2
b Institute of Advanced Biomedical Engineering and Science , Tokyo Women's Medical University , Tokyo 162-8666 , Japan.
3
c Nippi Research Institute of Biomatrix , Toride , Ibaraki 302-0017 , Japan.
4
d Department of Medical Education and Primary Care , Kyoto Prefectural University of Medicine , Kyoto 602-8566 , Japan.
5
e Department of Clinical and Pharmaceutical Sciences , Showa Pharmaceutical University , Tokyo 194-8543 , Japan.

Abstract

PURPOSE:

Our previous studies indicate that phorbol 12-myristate 13-acetate (PMA)-treated U937 cells cultured on collagen I -coated dishes express lowered production of pro-inflammatory mediators in parallel through reduced Reactive oxygen species (ROS) levels. By contrast, PMA-treated U937 cells on gelatin, the denatured collagen, show enhanced production of pro-inflammatory mediators, mediated by up-regulating autophagy levels. The present study is aimed to investigate the effect of ROS levels in PMA-treated U937 cells cultured on gelatin-coated surface.

MATERIAL AND METHODS:

MTT assay, flow cytometric analysis of ROS and autophagy, biochemical detection of antioxidant levels, ELISA assay, and western blot were used.

RESULTS:

Gelatin-coating increased ROS levels in PMA-treated U937 cells. Increased ROS levels are involved in the regulation of cell aggregation and the release of pro-inflammatory mediators in gelatin-coated culture. These results lead to the query about the crosstalk between the two positive regulators, the autophagy and ROS. Autophagy induction is attenuated by NAC treatment, but the treatment with autophagy inhibitor, 3-methyladenine, does not affect ROS levels, suggesting ROS are upstream of autophagy in the regulation axis of differentiated U937 cells on gelatin-coated surface. Further study confirmed that upregulation of autophagy was responsible for ROS-induced cell aggregation and production of pro-inflammatory mediators.

CONCLUSION:

The results suggest that gelatin-coating promotes the aggregation of PMA-treated U937 cells and the production of pro-inflammatory mediators by ROS-autophagy signaling pathway.

KEYWORDS:

Gelatin; ROS; autophagy; multicellular aggregates; pro-inflammatory mediators

Supplemental Content

Full text links

Icon for Taylor & Francis
Loading ...
Support Center