Long-term downregulation of Polo-like kinase 1 increases the cyclin-dependent kinase inhibitor p21<sup>WAF1/CIP1</sup>

Kreis, Nina-Naomi; Sommer, Katharina; Sanhaji, Mourad; Krämer, Andrea; Matthess, Yves; Kaufmann, M.; Strebhardt, Klaus; Yuan, Juping

doi:10.4161/cc.8.3.7651

Cited by 53 publications

(84 citation statements)

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“…Western blot analysis, cytosolic and nuclear extract separation Western blot analysis was carried out as previously described (Kreis et al, 2009). Mouse monoclonal antibodies against cyclin B1, Cdk1, Plk1, p53, p14ARF, MDM2, caspase-3, rabbit polyclonal antibodies against p53 and E6-AP, goat antibodies against HPV E6 and Aurora A, goat anti-mouse/ anti-rabbit and chicken anti-goat secondary antibodies were purchased from Santa Cruz (Heidelberg, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Mouse monoclonal antibodies against calnexin and lamin B1 were from BD Biosciences (Heidelberg, Germany) and Biozol, respectively. Western blots were quantified as described previously (Kreis et al, 2009). Cytosolic and nuclear extracts were prepared by using the Nuclear Complex Co-IP Kit (Active Motif, Rixensart, Belgium).…”

Section: Methodsmentioning

confidence: 99%

“…HeLa 776 clones with various levels of cyclin B1 were generated as described previously (Yuan et al, 2006). Cell synchronization to the G1/S boundary by double thymidine block and to prometaphase by thymidine/nocodazole treatment and cell lysis were carried out, as described (Kreis et al, 2009). Camptothecin, cycloheximide, RO-3306 and ZM447439 were from Fluka (Buchs, Switzerland), Sigma-Aldrich (Taufkirchen, Germany), Roche Applied Science (Mannheim, Germany) and Biozol (Eching, Germany), respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Camptothecin, cycloheximide, RO-3306 and ZM447439 were from Fluka (Buchs, Switzerland), Sigma-Aldrich (Taufkirchen, Germany), Roche Applied Science (Mannheim, Germany) and Biozol (Eching, Germany), respectively. CaSki stable cell clones expressing shRNA cyclin B1/GFP were established as described (Kreis et al, 2009).…”

Section: Methodsmentioning

confidence: 99%

“…Finally, it is well known that disruption of mitosis alone can promote the sensitivity of cancer cells to chemotherapeutics, which we have also observed (Yuan et al, 2006;Androic et al, 2008;Kreis et al, 2009). To address the function of p53 in this scenario, HeLa cells were depleted of either cyclin B1, p53 or both of them and then subjected to camptothecin.…”

Section: Dna Damage Response Of Hela 776-6 Cellsmentioning

confidence: 99%

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Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

et al. 2010

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Abrogation of functional p53 is responsible for malignant cell transformation and the maintenance of malignant state of human papillomavirus-infected cancer cells. Thus, restoration of p53 has been regarded as an important strategy for molecular intervention combating papillomavirus-associated malignancies. We show here that depleting cyclin B1 stabilizes and reactivates p53 in papillomavirusinfected cervical cancer cell lines HeLa and CaSki. HeLa cells depleted of cyclin B1 exhibit mitotic defects in spindle formation and chromosome alignment. Downregulation of cyclin B1 increases p14 alternative reading frame of p16, the positive regulator of p53, and decreases phosphorylation of Ser315 in p53. Whereas RO-3306, a selective inhibitor of cyclin-dependent kinase 1 (Cdk1), suppresses this phosphorylation at Ser315 of p53, ZM447439, targeting Aurora A/B kinases, shows no effect. Further analyses in HeLa cells and HCT116 p53(À/À) cells suggest that the Ser315 phosphorylation by Cdk1 regulates negatively the protein stability and the function of p53. Moreover, increased p53 in HeLa cells is functional by showing its increased downstream effectors p21, mouse double minute 2 and Bax. Restoration of p53 and silencing cyclin B1 render cervical carcinoma cells more susceptible to DNA damage agent camptothecin. Taken together, targeting cyclin B1 might be an attractive strategy for preventing and treating papillomavirus-associated cancer by reactivating p53 and by reducing the Cdk1 activity.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Dna Damage Response Of Hela 776-6 Cellsmentioning

confidence: 99%

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Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

et al. 2010

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RITA modulates cell migration and invasion by affecting focal adhesion dynamics

et al. 2019

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RITA, the RBP‐J interacting and tubulin‐associated protein, has been reported to be related to tumor development, but the underlying mechanisms are not understood. Since RITA interacts with tubulin and coats microtubules of the cytoskeleton, we hypothesized that it is involved in cell motility. We show here that depletion of RITA reduces cell migration and invasion of diverse cancer cell lines and mouse embryonic fibroblasts. Cells depleted of RITA display stable focal adhesions (FA) with elevated active integrin, phosphorylated focal adhesion kinase, and paxillin. This is accompanied by enlarged size and disturbed turnover of FA. These cells also demonstrate increased polymerized tubulin. Interestingly, RITA is precipitated with the lipoma‐preferred partner (LPP), which is critical in actin cytoskeleton remodeling and cell migration. Suppression of RITA results in reduced LPP and α‐actinin at FA leading to compromised focal adhesion turnover and actin dynamics. This study identifies RITA as a novel crucial player in cell migration and invasion by affecting the turnover of FA through its interference with the dynamics of actin filaments and microtubules. Its deregulation may contribute to malignant progression.

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Rescue of p53 functions by in vitro‐transcribed mRNA impedes the growth of high‐grade serous ovarian cancer

Raab,

Kostova,

Peña‐Llopis

et al. 2023

Cancer Communications

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BackgroundThe cellular tumor protein p53 (TP53) is a tumor suppressor gene that is frequently mutated in human cancers. Among various cancer types, the very aggressive high‐grade serous ovarian carcinoma (HGSOC) exhibits the highest prevalence of TP53 mutations, present in >96% of cases. Despite intensive efforts to reactivate p53, no clinical drug has been approved to rescue p53 function. In this study, our primary objective was to administer in vitro‐transcribed (IVT) wild‐type (WT) p53‐mRNA to HGSOC cell lines, primary cells, and orthotopic mouse models, with the aim of exploring its impact on inhibiting tumor growth and dissemination, both in vitro and in vivo.MethodsTo restore the activity of p53, WT p53 was exogenously expressed in HGSOC cell lines using a mammalian vector system. Moreover, IVT WT p53 mRNA was delivered into different HGSOC model systems (primary cells and patient‐derived organoids) using liposomes and studied for proliferation, cell cycle progression, apoptosis, colony formation, and chromosomal instability. Transcriptomic alterations induced by p53 mRNA were analyzed using RNA sequencing in OVCAR‐8 and primary HGSOC cells, followed by ingenuity pathway analysis. In vivo effects on tumor growth and metastasis were studied using orthotopic xenografts and metastatic intraperitoneal mouse models.ResultsReactivation of the TP53 tumor suppressor gene was explored in different HGSOC model systems using newly designed IVT mRNA‐based methods. The introduction of WT p53 mRNA triggered dose‐dependent apoptosis, cell cycle arrest, and potent long‐lasting inhibition of HGSOC cell proliferation. Transcriptome analysis of OVCAR‐8 cells upon mRNA‐based p53 reactivation revealed significant alterations in gene expression related to p53 signaling, such as apoptosis, cell cycle regulation, and DNA damage. Restoring p53 function concurrently reduces chromosomal instability within the HGSOC cells, underscoring its crucial contribution in safeguarding genomic integrity by moderating the baseline occurrence of double‐strand breaks arising from replication stress. Furthermore, in various mouse models, treatment with p53 mRNA reduced tumor growth and inhibited tumor cell dissemination in the peritoneal cavity in a dose‐dependent manner.ConclusionsThe IVT mRNA‐based reactivation of p53 holds promise as a potential therapeutic strategy for HGSOC, providing valuable insights into the molecular mechanisms underlying p53 function and its relevance in ovarian cancer treatment.

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Long-term downregulation of Polo-like kinase 1 increases the cyclin-dependent kinase inhibitor p21^WAF1/CIP1

Cited by 53 publications

References 49 publications

Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

RITA modulates cell migration and invasion by affecting focal adhesion dynamics

Rescue of p53 functions by in vitro‐transcribed mRNA impedes the growth of high‐grade serous ovarian cancer

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Long-term downregulation of Polo-like kinase 1 increases the cyclin-dependent kinase inhibitor p21WAF1/CIP1

Cited by 53 publications

References 49 publications

Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

Restoration of the tumor suppressor p53 by downregulating cyclin B1 in human papillomavirus 16/18-infected cancer cells

RITA modulates cell migration and invasion by affecting focal adhesion dynamics

Rescue of p53 functions by in vitro‐transcribed mRNA impedes the growth of high‐grade serous ovarian cancer

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Long-term downregulation of Polo-like kinase 1 increases the cyclin-dependent kinase inhibitor p21^WAF1/CIP1