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The Role of SHP1 as a Tumor Suppressor in Bladder Cancer Cell Lines
Shannon McNall, MD, Daniel Pohl, MD, Kailey Hooper, BS, Travis Sullivan, MS, Kimberly Rieger-Christ, PhD.
Lahey Hospital and Medical Center, Burlington, MA, USA.
BACKGROUND: Protein tyrosine phosphatase nonreceptor type 6, also known as SHP1, is a tyrosine phosphatase that serves as a signaling molecule and a tumor suppressor in certain cancers. Lower SHP1 expression in bladder tumors was associated with poorer overall survival in The Cancer Genome Atlas (TCGA) dataset. The role of SHP1 in bladder cancer continues to be investigated. We aim to assess SHP1 expression patterns and its impact on the epithelial-mesenchymal transition (EMT) phenotype in bladder cancer cell lines, as well as the mechanism by which SHP1 affects bladder cancer cellular activity.
METHODS: SHP1 levels were evaluated in 12 human bladder cancer cell lines. SHP1 knock-down and forced expression were performed in four cell lines using lentiviral transduction. SHP1 protein expression was evaluated via western blot. Downstream targets of SHP1, such as Akt, were also evaluated. Cell proliferation of the transduced cell lines was evaluated using the MT Cell Viability Assay. Likewise, cell migration and invasion were determined using Transwell assays, with or without Matrigel.
RESULTS: There was significantly lower SHP1 protein expression in highly invasive cell lines compared to poorly invasive (p<0.01). In vitro alterations in SHP1 expression significantly affected the EMT phenotype of bladder cancer cell lines. Knocking-down expression of SHP1 in poorly invasive cell lines led to increased proliferation, invasion and migration of the cells (p<0.05), and phosphorylated (activated) Akt was increased in these cell lines (p<0.05). Forced expression of SHP1 in highly invasive cell lines led to decreased proliferation, invasion and migration of the cells (p<0.05), and phosphorylated Akt was decreased in these cell lines (p<0.05).
CONCLUSIONS: In bladder cancer cell lines, lower SHP1 expression is associated with features of more aggressive cancer. This information supports the utility of further research into SHP1 as a prognostic biomarker for bladder cancer or a target for future treatments.
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