Knockdown of CKAP2 Inhibits Proliferation, Migration, and Aggregate Formation in Aggressive Breast Cancer

Santos, Alexsandro dos; Ouellete, Geneviève; Diorio, Caroline; Elowe, Sabine; Durocher, Francine

doi:10.3390/cancers14153759

Cited by 3 publications

(1 citation statement)

References 80 publications

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“…The copyright holder for this preprint this version posted October 27, 2023. ; https://doi.org/10.1101/2023.10.27.564280 doi: bioRxiv preprint segregation (16)(17)(18)(19). CKAP2 knock-down leads to spindle assembly defects (16) and CKAP2 overexpression has been associated with tumor development (20)(21)(22) and is negatively correlated with survival rates in cancer patients (23). Our recent in vitro reconstitution experiments have demonstrated that CKAP2 substantially increases microtubule nucleation and growth rates (24), however, whether and how CKAP2 regulates microtubule dynamics and chromosome segregation in cells has yet to be explored.…”

Section: Introductionmentioning

confidence: 99%

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

Paim,

Lopez-Jauregui,

McAlear

et al. 2023

Preprint

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Regulation of microtubule dynamics by microtubule-associated proteins (MAPs) is essential for mitotic spindle assembly and chromosome segregation. Altered microtubule dynamics, particularly increased microtubule growth rates, were found to be a contributing factor for the development of chromosomal instability, which potentiates tumorigenesis. The MAP XMAP215/CKAP5 is the only known microtubule growth factor, and whether other MAPs regulate microtubule growth in cells is unclear. Our recent in vitro reconstitution experiments have demonstrated that Cytoskeleton-Associated Protein 2 (CKAP2) increases microtubule nucleation and growth rates, and here we find that CKAP2 is also an essential microtubule growth factor in cells. By applying CRISPR-Cas9 knock-in and knock-out as well as microtubule plus-end tracking live cell imaging, we show that CKAP2 is a mitotic spindle protein that ensures faithful chromosome segregation by regulating microtubule growth. Live cell imaging of endogenously-labelled CKAP2 showed that it localizes to the spindle during mitosis, and rapidly shifts its localization to the chromatin upon mitotic exit before being degraded. Cells lacking CKAP2 display reduced microtubule growth rates and an increased proportion of chromosome segregation errors and aneuploidy that may be a result of an accumulation of kinetochore-microtubule mis-attachments. Microtubule growth rates and chromosome segregation fidelity can be rescued upon CKAP2 expression in knock-out cells, revealing a direct link between CKAP2 expression and microtubule dynamics. Our results unveil a role of CKAP2 in regulating microtubule growth in cells and provide a mechanistic explanation for the oncogenic potential of CKAP2 misregulation.

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Section: Introductionmentioning

confidence: 99%

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

Paim,

Lopez-Jauregui,

McAlear

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

Paim,

Lopez-Jauregui,

McAlear

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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Regulation of microtubule dynamics by microtubule-associated proteins (MAPs) is essential for mitotic spindle assembly and chromosome segregation. Altered microtubule dynamics, particularly increased microtubule growth rates, were found to be a contributing factor for the development of chromosomal instability, which potentiates tumorigenesis. The MAP XMAP215/CKAP5 is the only known microtubule growth factor, and whether other MAPs regulate microtubule growth in cells is unclear. Our recent in vitro reconstitution experiments have demonstrated that Cytoskeleton-Associated Protein 2 (CKAP2) increases microtubule nucleation and growth rates, and here, we find that CKAP2 is also an essential microtubule growth factor in cells. By applying CRISPR-Cas9 knock-in and knock-out (KO) as well as microtubule plus-end tracking live cell imaging, we show that CKAP2 is a mitotic spindle protein that ensures faithful chromosome segregation by regulating microtubule growth. Live cell imaging of endogenously labeled CKAP2 showed that it localizes to the spindle during mitosis and rapidly shifts its localization to the chromatin upon mitotic exit before being degraded. Cells lacking CKAP2 display reduced microtubule growth rates and an increased proportion of chromosome segregation errors and aneuploidy that may be a result of an accumulation of kinetochore–microtubule misattachments. Microtubule growth rates and chromosome segregation fidelity can be rescued upon ectopic CKAP2 expression in KO cells, revealing a direct link between CKAP2 expression and microtubule dynamics. Our results unveil a role of CKAP2 in regulating microtubule growth in cells and provide a mechanistic explanation for the oncogenic potential of CKAP2 misregulation.

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ZC3H13 Enhances the Malignancy of Cervical Cancer by Regulating m6A Modification of CKAP2

Zhang,

Chen,

Chen

et al. 2023

Crit Rev Immunol

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Sustained expression of zinc finger CCCH-type containing 13 (ZC3H13) in tumors is essential for cancer cell malignancy; however, our understanding of its clinical effects and mechanisms in cervical cancer (CC) is limited. In this study, we aimed to reveal the effect on CC progression of ZC3H13-mediated N6-methyladenosine (m6A) modification to stabilize cytoskeleton-associated protein 2 (CKAP2) expression. CC tissues and paired adjacent normal tissues were collected from 50 patients. qRT-PCR was used to clarify ZC3H13 and CKAP2 expression levels in the CC tissues. The functional roles of ZC3H13 and CKAP2 in CC were analyzed by detecting the changes in CC cell proliferation, migration, invasion, and tumor growth <i>in vivo</i>. The regulatory relationship between ZC3H13 and CKAP2 was investigated by confirming m6A modification levels and their expression correlation. ZC3H13 and CKAP2 were highly expressed in CC and linked with poor prognosis. We observed that ZC3H13 inhibition decreased CC cell proliferation, invasion, and migration, while its facilitation promoted CC cell malignancy. ZC3H13 mediated m6A modification of CKAP2 to enhance CKAP2 expression in CC cells. Furthermore, CKAP2 overexpression partially restored the malignant phenotypic promotion induced by ZC3H13 overexpression in CC cells. In summary, this study revealed that ZC3H13-mediating m6A modification of CKAP2 promotes CC development. This finding should be conducive to an understanding of the role of ZC3H13-m6A-CKAP2 in CC and should provide an effective therapeutic target for this cancer.

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Knockdown of CKAP2 Inhibits Proliferation, Migration, and Aggregate Formation in Aggressive Breast Cancer

Cited by 3 publications

References 80 publications

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

The spindle protein CKAP2 regulates microtubule dynamics and ensures faithful chromosome segregation

ZC3H13 Enhances the Malignancy of Cervical Cancer by Regulating m6A Modification of CKAP2

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