Abstract
In fibroblasts, beryllium salt causes activation of the p53 transcription factor and induction of a senescence-like state. It is not known whether Be2+ can affect the proliferation of cancer cells, which are generally unsusceptible to senescence. A172 glioblastoma and RKO colon carcinoma cell lines each have wildtype p53, so these cell types have the potential to be responsive to agents that activate p53. In A172 cells, BeSO4 produced a G0/G1-phase cell cycle arrest and increased expression of senescence-associated β-galactosidase, an enzymatic marker of senescence. BeSO4 caused phosphorylation of serine-15 of p53, accumulation of p53 protein, and expression of p21, the cyclin-dependent kinase inhibitor that is prominent during senescence. BeSO4 inhibited A172 growth with an IC50 = 4.7 μM in a 6-day proliferation assay. In contrast, BeSO4 had no effect on RKO cells, even though Be2+ uptake was similar for the two cell types. This differential responsiveness marks BeSO4 as a reagent capable of activating a separable branch of the p53 signaling network. A172 and RKO cells are known to exhibit p53-dependent upregulation of p21 in response to DNA damage. The RKO cells produced high levels of p21 when exposed to DNA damaging agents, yet failed to express p21 when treated with BeSO4. Conversely, BeSO4 did not cause DNA damage in A172 cells, yet it was a potent inducer of p21 expression. These observations indicate that the growth control pathway affected by BeSO4 is distinct from the DNA damage response pathway, even though both ultimately converge on p53 and p21.
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Acknowledgements
We thank Dr. Bruce Lehnert, Dr. David Ward, and Dr. Hokwon Cho for helpful discussions, and Ms. Swapna Mudireddy for pilot studies to evaluate the feasibility of quantifying intracellular beryllium by ICP-MS. This work was funded by National Institutes of Health grant P20 RR-016464 from the INBRE Program of the National Center for Research Resources.
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10534_2010_9352_MOESM1_ESM.tif
Fig. S1: Total cell number declines over time at a logarithmic rate in A172 cells treated with a cytotoxic agent (etoposide). Mean ± standard deviation, error bars are not visible because they are narrower than the filled circle symbol (TIFF 76 kb)
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Gorjala, P., Gary, R.K. Beryllium sulfate induces p21CDKN1A expression and a senescence-like cell cycle arrest in susceptible cancer cell types. Biometals 23, 1061–1073 (2010). https://doi.org/10.1007/s10534-010-9352-y
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DOI: https://doi.org/10.1007/s10534-010-9352-y