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Involvement of NYD‐SP15 in growth and oxidative‐stress responses of ARPE‐19
Author(s) -
Xu Yidan,
Wang Ling,
Cao Liu,
Chen Lixun,
Liu Qinghuai
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.27104
Subject(s) - downregulation and upregulation , chemistry , cell growth , microbiology and biotechnology , transfection , oxidative stress , protein kinase b , apoptosis , gene knockdown , mapk/erk pathway , signal transduction , biology , biochemistry , gene
The aim of this study was to investigate the role of NYD‐SP15 in the growth and oxidative‐stress responses of ARPE‐19 cells. ARPE‐19 cell lines overexpressing wild type or RNA interference against NYD‐SP15 were established via lentivirus transfection. Cell growth and proliferation, migration, apoptosis, and cell cycle progression were monitored using the Cell Counting Kit‐8 assay, the wound scratch assay, and flow cytometry, respectively. Caspase‐3/8/9 activity was examined using the caspase‐3/8/9 assay kit. An hydrogen peroxide (H 2 O 2 )–induced oxidative‐stress damage model was used to study the effect of NYD‐SP15 knockdown by examining the activity of reactive oxygen species (ROS). Expressions of Kelch‐like ECH‐associated protein 1 (Keap‐1)/heme oxygenase‐1 (HO‐1)/nuclear factor erythroid 2–related factor 2 (Nrf2), mitogen‐activated protein kinase (MAPK), and Akt were detected by Western blot analysis. The mRNA chip of NYD‐SP15 overexpressed ARPE‐19 cells as well as controls were performed by one array plus process. Overexpression (OE) of NYD‐SP15 inhibited the proliferation and migration of ARPE‐19 cells, and led to apoptosis and caspase‐3/9 activation. OE of NYD‐SP15 inhibited MAPKs and Akt signaling. Downregulation of NYD‐SP15 had no effect on the growth of normally cultured ARP19 cells with 10% fetal bovine serum, but promoted the growth of ARP19 cells in the presence of starvation challenge. Gene chip showed that OE of NYD‐SP15 led to downregulation of 254 genes and upregulation of 57 genes. Downregulation of NYD‐SP15 also exerted a protective effect on H 2 O 2 ‐induced cell apoptosis and ROS. NYD‐SP15 downregulation led to increments in the expression of Nrf2, Keap‐1, and HO‐1 in response to 200 μM H 2 O 2 . NYD‐SP15 might inhibit the growth, proliferation, and migration and promote apoptosis of ARPE‐19 cells via MAPK and Akt signaling. Downregulation of NYD‐SP15 could protect ARPE‐19 cells from H 2 O 2 ‐induced oxidative damage by active Keap‐1/HO‐1/Nrf2, Akt, and MAPK signaling.