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Coiled-coil domain containing 3 suppresses breast cancer growth by protecting p53 from proteasome-mediated degradation

Abstract

Coiled-coil domain containing 3 (CCDC3) was previously shown to regulate liver lipid metabolism as a secretory protein. Here, we report an unexpected intracellular role of CCDC3 as a tumor suppressor in breast cancer (BrC). Bioinformatics datasets analysis showed that CCDC3 is under-expressed in BrCs, while its higher levels are correlated with higher overall survival and lower relapse of cancer patients, and CCDC3 is positively correlated with p53 and its target genes. Ectopic CCDC3 markedly suppressed proliferation, colony formation, and xenograft tumor growth by augmenting p53 activity in BrC cells. Depletion of endogenous CCDC3 by CRISPR-Cas9 increased proliferation and drug resistance of BrC cells by alleviating 5-Fluorouracil (5-FU)-induced p53 level and activity. Mechanistically, CCDC3 bound to the C-termini of p53 and MDM2, consequently stabilizing p53 in the nucleus and impairing MDM2 recruitment of p53 to the 26S proteosome without inhibiting p53 ubiquitination. p53 induced CCDC3 expression by binding to its promoter in BrC cells. Our results unveil a unique mechanism underlying CCDC3 activation of p53 in a positive feedback fashion to suppress BrC growth.

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Fig. 1: CCDC3 is down-regulated and positively correlated with p53 in breast cancers.
Fig. 2: CCDC3 suppresses the growth of breast cancer cells and breast tumors, while CCDC3 deficiency increases proliferation of Cal51 cells.
Fig. 3: CCDC3 stabilizes and activates wt p53.
Fig. 4: CCDC3 leads to accumulation of ubiquitinated wt p53 and MDM2.
Fig. 5: CCDC3 stabilizes p53 in the nucleus.
Fig. 6: CCDC3 interacts with p53 and MDM2 via their C-termini.
Fig. 7: CCDC3 binds to p53 in the nucleus and reduces the binding of p53 to S6b.
Fig. 8: CCDC3 is a transcriptional target of p53.

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Acknowledgements

We thank James Jackson, Wesley D. Frey, Joe Gray, and Jean-Paul Borg for offering some BrC cell lines (Cal 51, MDA-MB-175, MPE600, SKBR7) and the CRISPR-p53KO-mCherry construct (James Jackson) and Heather Machado for sharing her animal protocol (Mammary fat pad injection). HL and SXZ were supported in part by NIH-NCI grants R01CA095441, R01CA234605, and R01CA127724 as well as R21CA272890. HL was also in part supported by the Reynolds and Ryan Families Endowed Chair fund and the NIH-NCI grant U01CA252965.

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CYL and HML equally conducted the majority of the experiments, analyzed the results and organized them into figures under supervision of SXZ and HL; JHJ initiated the study and conducted some experiments on regulation of p53 by CCDC3; JQW and AS assisted in the animal experiment; CL assisted in some in vitro assay; YWZ assisted in the animal protocol; RLS assisted in cellular fractionation; SXZ conducted some in vitro and animal experiments with HML and AS; SXZ and HL instructed the whole project; CYL and HL composed the manuscript; CYL, HML, SXZ, and HL revised the manuscript.

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Correspondence to Shelya X. Zeng or Hua Lu.

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Li, C., Lee, H., Jung, J.H. et al. Coiled-coil domain containing 3 suppresses breast cancer growth by protecting p53 from proteasome-mediated degradation. Oncogene 42, 154–164 (2023). https://doi.org/10.1038/s41388-022-02541-1

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