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Targeting aurora kinases limits tumour growth through DNA damage-mediated senescence and blockade of NF-κB impairs this drug-induced senescence.

Liu Y, Hawkins OE, Su Y, Vilgelm AE, Sobolik T, Thu YM, Kantrow S, Splittgerber RC, Short S, Amiri KI, Ecsedy JA, Sosman JA, Kelley MC, Richmond A - EMBO Mol Med (2012)

Bottom Line: Mechanistic analyses revealed that inhibition of aurora kinases induced polyploidy and the ATM/Chk2 DNA damage response, which mediated senescence and a NF-κB-related, senescence-associated secretory phenotype (SASP).Blockade of IKKβ/NF-κB led to reversal of MLN8237-induced senescence and SASP.Altogether, these data demonstrate that induction of senescence, coupled with immune surveillance, can limit melanoma growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterans Affairs, Tennessee Valley Healthcare System, Nashville, TN, USA.

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Inhibition of aurora kinases induces cellular senescence in vitro independent of p53Hs294T (p53WT), SK-Mel-2 (p53 mutant), SK-Mel-5 (p53WT) and SK-Mel-28 (p53 mutant) cells were treated with 1 µM MLN8237 for 6 or 12 days. After treatment, viable cells were counted and compared to the initial cell number. Data indicate mean values ± SD (n = 3) from one representative of three independent experiments. p-value shown represents difference between treated group and day 0 control group (Student's t-test).Cultures of Hs294T, SK-Mel-2, SK-Mel-5 and SK-Mel-28 cells were treated with MLN8237 or vehicle for indicated time points and apoptosis was analysed by FACS analysis for propidium iodide (PI) and Annexin V staining. Data indicate mean values ± SD (n = 3) from triplicate experiments.Hs294T cells were treated with 1 µM MLN8237 or vehicle for 5 days and the change in morphology was captured with an AxioVision microscope.Hs294T cells were treated with 1 µM MLN8237 for 5 days, and senescence was determined by β-galactosidase staining.Melanoma cell lines with wild-type p53 (Hs294T and SK-Mel-5) or mutated p53 (SK-Mel-2 and SK-Mel-28) were treated with 1 µM or 5 µM MLN8237 for 5 days. After treatment, p53, p63, p73, p21, p16 and GAPDH were analysed by Western blot.Hs294T and SK-Mel-28 cells were treated with 1 µM MLN8237 or vehicle in the presence or absence of the p53 inhibitor pifithrin-α (PTF-α, 5 µM) or DMSO for 5 days. After treatment, β-galactosidase staining was performed. All experiments were conducted at least three times independently with reproducible results.
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fig02: Inhibition of aurora kinases induces cellular senescence in vitro independent of p53Hs294T (p53WT), SK-Mel-2 (p53 mutant), SK-Mel-5 (p53WT) and SK-Mel-28 (p53 mutant) cells were treated with 1 µM MLN8237 for 6 or 12 days. After treatment, viable cells were counted and compared to the initial cell number. Data indicate mean values ± SD (n = 3) from one representative of three independent experiments. p-value shown represents difference between treated group and day 0 control group (Student's t-test).Cultures of Hs294T, SK-Mel-2, SK-Mel-5 and SK-Mel-28 cells were treated with MLN8237 or vehicle for indicated time points and apoptosis was analysed by FACS analysis for propidium iodide (PI) and Annexin V staining. Data indicate mean values ± SD (n = 3) from triplicate experiments.Hs294T cells were treated with 1 µM MLN8237 or vehicle for 5 days and the change in morphology was captured with an AxioVision microscope.Hs294T cells were treated with 1 µM MLN8237 for 5 days, and senescence was determined by β-galactosidase staining.Melanoma cell lines with wild-type p53 (Hs294T and SK-Mel-5) or mutated p53 (SK-Mel-2 and SK-Mel-28) were treated with 1 µM or 5 µM MLN8237 for 5 days. After treatment, p53, p63, p73, p21, p16 and GAPDH were analysed by Western blot.Hs294T and SK-Mel-28 cells were treated with 1 µM MLN8237 or vehicle in the presence or absence of the p53 inhibitor pifithrin-α (PTF-α, 5 µM) or DMSO for 5 days. After treatment, β-galactosidase staining was performed. All experiments were conducted at least three times independently with reproducible results.

Mentions: Although previous reports demonstrated that blocking AURKA/B using small-molecule inhibitors induces widespread apoptosis in different types of human cancer (Dar et al, 2008; Gorgun et al, 2010), we observed little apoptosis in MLN8054/MLN8237-treated melanoma patient tumour implants (Supporting Information Fig S9). In vitro studies showed that while the treatment with MLN8237 markedly reduced the number of viable cells (Fig 2A), it induced apoptosis in only 25% of SK-Mel-2 cells and in <10% of cells in three other melanoma cell lines (Fig 2B).


Targeting aurora kinases limits tumour growth through DNA damage-mediated senescence and blockade of NF-κB impairs this drug-induced senescence.

Liu Y, Hawkins OE, Su Y, Vilgelm AE, Sobolik T, Thu YM, Kantrow S, Splittgerber RC, Short S, Amiri KI, Ecsedy JA, Sosman JA, Kelley MC, Richmond A - EMBO Mol Med (2012)

Inhibition of aurora kinases induces cellular senescence in vitro independent of p53Hs294T (p53WT), SK-Mel-2 (p53 mutant), SK-Mel-5 (p53WT) and SK-Mel-28 (p53 mutant) cells were treated with 1 µM MLN8237 for 6 or 12 days. After treatment, viable cells were counted and compared to the initial cell number. Data indicate mean values ± SD (n = 3) from one representative of three independent experiments. p-value shown represents difference between treated group and day 0 control group (Student's t-test).Cultures of Hs294T, SK-Mel-2, SK-Mel-5 and SK-Mel-28 cells were treated with MLN8237 or vehicle for indicated time points and apoptosis was analysed by FACS analysis for propidium iodide (PI) and Annexin V staining. Data indicate mean values ± SD (n = 3) from triplicate experiments.Hs294T cells were treated with 1 µM MLN8237 or vehicle for 5 days and the change in morphology was captured with an AxioVision microscope.Hs294T cells were treated with 1 µM MLN8237 for 5 days, and senescence was determined by β-galactosidase staining.Melanoma cell lines with wild-type p53 (Hs294T and SK-Mel-5) or mutated p53 (SK-Mel-2 and SK-Mel-28) were treated with 1 µM or 5 µM MLN8237 for 5 days. After treatment, p53, p63, p73, p21, p16 and GAPDH were analysed by Western blot.Hs294T and SK-Mel-28 cells were treated with 1 µM MLN8237 or vehicle in the presence or absence of the p53 inhibitor pifithrin-α (PTF-α, 5 µM) or DMSO for 5 days. After treatment, β-galactosidase staining was performed. All experiments were conducted at least three times independently with reproducible results.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3569660&req=5

fig02: Inhibition of aurora kinases induces cellular senescence in vitro independent of p53Hs294T (p53WT), SK-Mel-2 (p53 mutant), SK-Mel-5 (p53WT) and SK-Mel-28 (p53 mutant) cells were treated with 1 µM MLN8237 for 6 or 12 days. After treatment, viable cells were counted and compared to the initial cell number. Data indicate mean values ± SD (n = 3) from one representative of three independent experiments. p-value shown represents difference between treated group and day 0 control group (Student's t-test).Cultures of Hs294T, SK-Mel-2, SK-Mel-5 and SK-Mel-28 cells were treated with MLN8237 or vehicle for indicated time points and apoptosis was analysed by FACS analysis for propidium iodide (PI) and Annexin V staining. Data indicate mean values ± SD (n = 3) from triplicate experiments.Hs294T cells were treated with 1 µM MLN8237 or vehicle for 5 days and the change in morphology was captured with an AxioVision microscope.Hs294T cells were treated with 1 µM MLN8237 for 5 days, and senescence was determined by β-galactosidase staining.Melanoma cell lines with wild-type p53 (Hs294T and SK-Mel-5) or mutated p53 (SK-Mel-2 and SK-Mel-28) were treated with 1 µM or 5 µM MLN8237 for 5 days. After treatment, p53, p63, p73, p21, p16 and GAPDH were analysed by Western blot.Hs294T and SK-Mel-28 cells were treated with 1 µM MLN8237 or vehicle in the presence or absence of the p53 inhibitor pifithrin-α (PTF-α, 5 µM) or DMSO for 5 days. After treatment, β-galactosidase staining was performed. All experiments were conducted at least three times independently with reproducible results.
Mentions: Although previous reports demonstrated that blocking AURKA/B using small-molecule inhibitors induces widespread apoptosis in different types of human cancer (Dar et al, 2008; Gorgun et al, 2010), we observed little apoptosis in MLN8054/MLN8237-treated melanoma patient tumour implants (Supporting Information Fig S9). In vitro studies showed that while the treatment with MLN8237 markedly reduced the number of viable cells (Fig 2A), it induced apoptosis in only 25% of SK-Mel-2 cells and in <10% of cells in three other melanoma cell lines (Fig 2B).

Bottom Line: Mechanistic analyses revealed that inhibition of aurora kinases induced polyploidy and the ATM/Chk2 DNA damage response, which mediated senescence and a NF-κB-related, senescence-associated secretory phenotype (SASP).Blockade of IKKβ/NF-κB led to reversal of MLN8237-induced senescence and SASP.Altogether, these data demonstrate that induction of senescence, coupled with immune surveillance, can limit melanoma growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterans Affairs, Tennessee Valley Healthcare System, Nashville, TN, USA.

Show MeSH
Related in: MedlinePlus