Regulation of miRNA-21 by reactive oxygen species-activated ERK/NF-κB in arsenite-induced cell transformation
Graphical abstract
Highlights
► Chronic exposure to arsenite induces the transformation of HELF cells. ► Arsenite improves miR-21 expression by reactive oxygen species activation of the ERK/NF-κB pathway. ► NF-κB regulates miR-21 expression by binding directly to the promoter of miR-21. ► MiR-21 has a feedback effect in regulating ERK activation via Spry1. ► MiR-21 affects the malignancy potential of arsenite-transformed HELF cells.
Section snippets
Cell culture
Immortalized HELF cells were obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences (Shanghai, China). These cells are human sarcoma virus 40 immortalized, nontumorigenic, diploid fibroblasts from the lungs of hysterotomy-derived embryos [19]. Cells were maintained in 5% CO2 at 37 °C in Dulbecco's modified Eagle medium (DMEM; Life Technologies/Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Life Technologies/Gibco), penicillin (100 U/ml),
Upregulation of miR-21 and activation of ERK and NF-κB induced by arsenite in HELF cells
To determine if arsenite increases transformation, HELF cells were incubated with or without 1.0 μM arsenite continuously. After 30 passages (about 15 weeks) of exposure, arsenite was removed from the cell culture medium before the initiation of experiments. Anchorage-independent growth is a characteristic gained by cancer cells conferring mobility without the consequence of apoptosis [18]. Colonies formed from arsenite-exposed cells were both larger (Supplementary Fig. S2A) and greater in number
Discussion
Recent mechanistic studies of arsenite carcinogenesis have directly or indirectly shown the involvement of altered epigenetic regulation in gene expression changes induced by arsenite exposure [34]. Arsenite is a well-established human carcinogen [35], [36]. A close association and positive correlation exists between arsenite exposure and increased incidences of various forms of cancer [7], [35]. Although various arsenite-regulated genes and processes have been described, many of the molecular
Acknowledgments
The authors thank Donald L. Hill (University of Alabama at Birmingham, USA) for editing. This work was supported by the Natural Science Foundations of China (30872146 and 81072327), the Research Fund for the Doctoral Program of Higher Education of China (20103234110005), the Key Program of the Educational Commission of Jiangsu Province of China (11KJA330002), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (2010).
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These authors contributed equally to this work.