Journal article
Microcystin-LR activates serine/threonine kinases and alters the phosphoproteome in human HepaRG cells
Toxicon (Oxford), Vol.249, p.108072
10/01/2024
PMID: 39154757
Abstract
Microcystin-LR (MCLR) exposure has been associated with development of hepatocellular carcinoma (HCC). Many of the carcinogenic mechanisms for MCLR have been attributed to the induction of cell survival and proliferation through altered protein phosphorylation pathways by inhibition of protein phosphatases 1 (PP1) and PP2A. The current study determined MCLR effects on the phosphoproteome in human HepaRG cells. Differentiated HepaRG cells were treated with either vehicle or MCLR followed by phosphoproteomic analysis and Western blotting of MAPK-activated proteins. MCLR decreased cell viability at 24 h at doses as low as 0.03 mu M. MCLR also caused a dose-dependent increase in phosphorylation of signaling and stress kinases. The number of decreased phosphosites by 0.1 mu M MCLR was similar between the 2 h (212) and 24 h (154) timepoints. In contrast, a greater number of phosphosites were increased at 24 h (567) versus the 2 h timepoint (136), indicating the hyperphosphorylation state caused by MCLR-mediated inhibition of PPs is time-dependent. A kinase perturbation analysis predicted that MCLR exposure at both 2 h and 24 h increased the function of aurora kinase B (AURKB), checkpoint kinase 1 (CHEK1), and serum and glucocorticoid-regulated kinase 1 (SGK1). STRING database analysis of the phosphosites altered by MCLR exposure revealed pathways associated with cell proliferation and survival, including ribosomal protein S6 kinase (RSK), and vascular endothelial growth factor receptor (VEGFR2)-mediated vascular permeability. In addition, several cancer-related KEGG pathways were enriched at both 2 h and 24 h timepoints, and multiple cancer-related disease-gene associations were identified at the 24 h timepoint. Many of the kinases and pathways described above play crucial roles in the development of HCC by affecting processes such as invasion and metastasis. Overall, our data indicate that MCLR-mediated changes in protein phosphorylation involve biological pathways related to carcinogenesis that may contribute to the development of HCC.
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Details
- Title
- Microcystin-LR activates serine/threonine kinases and alters the phosphoproteome in human HepaRG cells
- Creators
- Victor O. Ikumawoyi - Washington State University, Department of Pharmaceutical SciencesKatherine D. Lynch - Washington State University SpokaneDayne T. Iverson - Washington State University SpokaneM. Ridge Call - Washington State Univ, Dept Pharmaceut Sci, Spokane, WA 99202 USAGuihua Eileen Yue - Washington State University, Department of Pharmaceutical SciencesBhagwat Prasad - Washington State University, Department of Pharmaceutical SciencesJohn D. Clarke - Washington State University, Department of Pharmaceutical Sciences
- Publication Details
- Toxicon (Oxford), Vol.249, p.108072
- Academic Unit
- Department of Pharmaceutical Sciences
- Publisher
- Elsevier
- Number of pages
- 14
- Grant note
- Washington State University R01ES032558 / National Institute of Environmental Health Sciences; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Environmental Health Sciences (NIEHS)
- Identifiers
- 99901181237001842
- Language
- English
- Resource Type
- Journal article