A new, unexpected action of olomoucine, a CDK inhibitor, on normal human cells: Up‐regulation of CLIMP‐63, a cytoskeleton‐linking membrane protein

Węsierska‐Gądek, Józefa; Gueorguieva, Marieta; Kramer, Matthias P.; Ranftler, Carmen; Sarg, Bettina; Lindner, Herbert

doi:10.1002/jcb.21596

Cited by 10 publications

(9 citation statements)

References 51 publications

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“…Its cell cycle inhibitory effect strongly differs between cancer and normal cells. Whereas the exponentially growing human MCF‐7 breast cancer cells, which show a high mitotic potential, are arrested in G 2 phase of the cell cycle, 6 ROSC has almost no effect on the cell cycle of normal MRC‐5 human embryo fibroblasts, 8 which have only a very low mitotic index. Moreover, ROSC has stronger antiproliferative action in MCF‐7 than in MRC‐5 cells.…”

Section: Introductionmentioning

confidence: 99%

Roscovitine Differentially Affects Asynchronously Growing and Synchronized Human MCF‐7 Breast Cancer Cells

Maurer

Komina

Węsierska‐Gądek

2009

Annals of the New York Academy of Sciences

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Roscovitine (ROSC), a selective blocker of cyclin-dependent kinases (CDKs) efficiently inhibits proliferation of exponentially growing human MCF-7 breast cancer cells by induction of cell cycle arrest and p53-mediated apoptosis. ROSC blocks MCF-7 cells in G(2) phase in a time- and concentration-dependent manner. However, ROSC exerts a much weaker antiproliferative effect on human diploid fibroblasts. Therefore, in this study we questioned whether and to what extent the antiproliferative effect of ROSC depends on the cell cycle status of cancer cells exposed to the drug. We investigated the action of ROSC on asynchronous, exponentially growing, and on synchronized human MCF-7 breast cancer cells. MCF-7 cells were arrested in G(1) phase after serum withdrawal and in S phase by hydroxyurea. After serum refeeding, synchronized cells started to reenter the active cell cycle after 12 h. Exposure of G(1)-synchronized cells to ROSC prolonged the cell cycle arrest and was accompanied by a decrease in S-phase cells after 24 h. A similar but weaker trend occurred after ROSC administration, to cells released from G(1) arrest for 4 h. ROSC diminished the frequency of S-phase cells. Exposure of MCF-7 cells released from G(1) arrest to ROSC for 24 h resulted in an increase of the G(1)-cell population by 20%. Exposure to ROSC of MCF-7 cells released from S-phase block increased the ratio of S-phase cells. These results indicate that the cell cycle status of cancer cells prior to the onset of therapy determines the outcome of treatment.

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Section: Introductionmentioning

confidence: 99%

Roscovitine Differentially Affects Asynchronously Growing and Synchronized Human MCF‐7 Breast Cancer Cells

Maurer

Komina

Węsierska‐Gądek

2009

Annals of the New York Academy of Sciences

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“…OLO, a substituted purine analogue, is an approximately 10‐ to 20‐fold weaker CDK inhibitor than ROSC. OLO has almost no effect on the proliferation and cell cycle progression of human diploid MRC‐5 fibroblasts 23 but strongly affects the viability of human HL‐60 cancer cells. Because MRC‐5 cells exhibit a low mitotic index and the reduced ratio of S phase compared with HL‐60 cells, the question of what might determine the sensitivity of cells to OLO arose: primarily the proliferative status of the treated cells or perhaps other endogenous factors?…”

Section: Discussionmentioning

confidence: 99%

“…It has been recently reported that OLO did not affect the viability of normal human MRC‐5 fibroblasts, but it inhibited the proliferation of human HL‐60 leukemia cells. After exposure of HL‐60 cells to OLO at a final concentration of 150 μmol/L for 24 h, the number of living cells was markedly reduced 23 . However, the exact mechanism by which leukemic cells were eliminated has not been shown.…”

Section: Introductionmentioning

confidence: 99%

Is Olomoucine, a Weak CDK2 Inhibitor, Able to Induce Apoptosis in Cancer Cells?

Wandl

Węsierska‐Gądek

2009

Annals of the New York Academy of Sciences

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Olomoucine (OLO), a substituted purine analogue, is a much weaker inhibitor of cyclin-dependent kinases than other closely related ATP derivatives. It has been recently reported that OLO did not affect the viability of normal human MRC-5 fibroblasts, but it inhibited the proliferation of human HL-60 leukemia cells. Therefore, it was interesting to explore the antiproliferative effect of OLO and to characterize its action on distinct human cancer cells differing in the functional status of the cell cycle. Human HeLa cervical carcinoma and HL-60 leukemia cells were continuously exposed to increasing concentrations of OLO for 24 h and 48 h or alternatively, cells after treatment for 24 h were postincubated in a drug-free medium. Surprisingly, OLO more strongly affected the proliferation of HL-60 cells than that of HeLa cells. Flow cytometric analyses revealed that OLO at higher doses increases the frequency of a hypoploid HL-60 cell population representing cells undergoing apoptosis. These results substantiated the data of the determination of the number of viable cells. Moreover, OLO at higher doses modulates the cell cycle progression of tested cancer cells. Detailed analyses of the DNA concentration in single cells revealed that OLO-mediated reduction of the number of G(1)-phase cells was accompanied by an increase of the frequency of G(2)-phase cells. The kinetics of these changes differed between both tested cancer cell lines, suggesting that some cancer cells exhibit increased susceptibility to OLO action. It remains to clarify whether the strong proapoptotic effect of OLO observed in HL-60 cells depends on their differentiation status.

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“…Olomoucine is also a 2,6,9-trisubstituted purine [1]. Olomoucine displays lower pharmacological potency than roscovitine but a similar, though not identical, activity profile [7]. Interestingly, olomoucine has an inactive isomer, named iso-olomoucine that becomes the ideal negative control for biological experiments [8,9].…”

Section: Introductionmentioning

confidence: 98%

Transcriptional modulation of apoptosis regulators by roscovitine and related compounds

et al. 2011

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Chemical inhibitors of cyclin-dependent kinase (CDK), like roscovitine, are promising drugs in the context of new cancer therapies. Roscovitine and related compounds, like seliciclib and olomoucine, are effective inducers of apoptosis in many proliferating cells in culture. These compounds are known to activate the intrinsic or mitochondrial pathway of apoptosis. In order to better characterize this intrinsic pathway, a transcriptional analysis was performed using the reverse transcriptase-multiplex ligation-dependent probe amplification procedure (RT-MLPA). In five cell lines, we detected an early and marked reduction of most transcripts, which is consistent with the disruption of transcription that results from the inhibition of CDK7 and CDK9. However, the mRNA of p53-upregulated modulator of apoptosis (PUMA) gene escaped from this transcription inhibition in neuroblastoma cells with a functional p53 protein. The increase of PUMA mRNA was not found in roscovitine-treated cell lines defective in p53, which underwent apoptosis like their p53 proficient counterparts. In addition, in SH-SY5Y cells, sublethal and lethal concentrations of roscovitine produced equivalent increases of PUMA mRNA and protein. In conclusion, the increased expression of PUMA was not associated with apoptosis induction. On the contrary, mRNA and protein depletion of MCL-1 gene correlated the best with cell demise. Moreover, NOXA protein suffered a far minor decrease than MCL-1. Because of the selective neutralization of NOXA by MCL-1, we hypothesize that the disruption of this balance is a critical event in apoptosis induction by roscovitine and related compounds.

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A new, unexpected action of olomoucine, a CDK inhibitor, on normal human cells: Up‐regulation of CLIMP‐63, a cytoskeleton‐linking membrane protein

Cited by 10 publications

References 51 publications

Roscovitine Differentially Affects Asynchronously Growing and Synchronized Human MCF‐7 Breast Cancer Cells

Roscovitine Differentially Affects Asynchronously Growing and Synchronized Human MCF‐7 Breast Cancer Cells

Is Olomoucine, a Weak CDK2 Inhibitor, Able to Induce Apoptosis in Cancer Cells?

Transcriptional modulation of apoptosis regulators by roscovitine and related compounds

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