CDC20 Holds Novel Regulation Mechanism in RPA1 During Different Stages of DNA Damage to Induce Radio-Chemoresistance

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 Aug 10

PMID 39125953

Authors

Yang Gao

Pengbo Wen

Chenran Shao

Cheng Ye

Yuji Chen

Junyu You

Zhongjing Su

Affiliations

Soon will be listed here.

Abstract

Targeting CDC20 can enhance the radiosensitivity of tumor cells, but the function and mechanism of CDC20 on DNA damage repair response remains vague. To examine that issue, tumor cell lines, including KYSE200, KYSE450, and HCT116, were utilized to detect the expression, function, and underlying mechanism of CDC20 in radio-chemoresistance. Western blot and immunofluorescence staining were employed to confirm CDC20 expression and location, and radiation could upregulate the expression of CDC20 in the cell nucleus. The homologous recombination (HR) and non-homologous end joining (NHEJ) reporter gene systems were utilized to explore the impact of CDC20 on DNA damage repair, indicating that CDC20 could promote HR repair and radio/chemo-resistance. In the early stages of DNA damage, CDC20 stabilizes the RPA1 protein through protein-protein interactions, activating the ATR-mediated signaling cascade, thereby aiding in genomic repair. In the later stages, CDC20 assists in the subsequent steps of damage repair by the ubiquitin-mediated degradation of RPA1. CCK-8 and colony formation assay were used to detect the function of CDC20 in cell vitality and proliferation, and targeting CDC20 can exacerbate the increase in DNA damage levels caused by cisplatin or etoposide. A tumor xenograft model was conducted in BALB/c-nu/nu mice to confirm the function of CDC20 in vivo, confirming the in vitro results. In conclusion, this study provides further validation of the potential clinical significance of CDC20 as a strategy to overcome radio-chemoresistance via uncovering a novel role of CDC20 in regulating RPA1 during DNA damage repair.

References

He W, Meng J . CDC20: a novel therapeutic target in cancer. Am J Transl Res. 2023; 15(2):678-693. PMC: 10006751. View

Zhang Y, Wu L, Wang Z, Wang J, Roychoudhury S, Tomasik B . Replication Stress: A Review of Novel Targets to Enhance Radiosensitivity-From Bench to Clinic. Front Oncol. 2022; 12:838637. PMC: 9305609. DOI: 10.3389/fonc.2022.838637. View

Bray F, Laversanne M, Sung H, Ferlay J, Siegel R, Soerjomataram I . Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024; 74(3):229-263. DOI: 10.3322/caac.21834. View

Campos A, Clemente-Blanco A . Cell Cycle and DNA Repair Regulation in the Damage Response: Protein Phosphatases Take Over the Reins. Int J Mol Sci. 2020; 21(2). PMC: 7014277. DOI: 10.3390/ijms21020446. View

Jeong S, Bui Q, Kwak M, Lee J, Lee P . Targeting Cdc20 for cancer therapy. Biochim Biophys Acta Rev Cancer. 2022; 1877(6):188824. DOI: 10.1016/j.bbcan.2022.188824. View

Larsen B, Benada J, Yung P, Bell R, Pappas G, Urban V . Cancer cells use self-inflicted DNA breaks to evade growth limits imposed by genotoxic stress. Science. 2022; 376(6592):476-483. DOI: 10.1126/science.abi6378. View

Volonte D, Sedorovitz M, Galbiati F . Impaired Cdc20 signaling promotes senescence in normal cells and apoptosis in non-small cell lung cancer cells. J Biol Chem. 2022; 298(10):102405. PMC: 9490043. DOI: 10.1016/j.jbc.2022.102405. View

Liu X, Song M, Wang P, Zhao R, Chen H, Zhang M . Targeted therapy of the AKT kinase inhibits esophageal squamous cell carcinoma growth in vitro and in vivo. Int J Cancer. 2019; 145(4):1007-1019. PMC: 6618024. DOI: 10.1002/ijc.32285. View

Valente D, Gentileschi M, Guerrisi A, Bruzzaniti V, Morrone A, Soddu S . Factors to Consider for the Correct Use of γH2AX in the Evaluation of DNA Double-Strand Breaks Damage Caused by Ionizing Radiation. Cancers (Basel). 2022; 14(24). PMC: 9776434. DOI: 10.3390/cancers14246204. View

10.

Qiu Z, Oleinick N, Zhang J . ATR/CHK1 inhibitors and cancer therapy. Radiother Oncol. 2017; 126(3):450-464. PMC: 5856582. DOI: 10.1016/j.radonc.2017.09.043. View

11.

Novo N, Romero-Tamayo S, Marcuello C, Boneta S, Blasco-Machin I, Velazquez-Campoy A . Beyond a platform protein for the degradosome assembly: The Apoptosis-Inducing Factor as an efficient nuclease involved in chromatinolysis. PNAS Nexus. 2023; 2(2):pgac312. PMC: 9944232. DOI: 10.1093/pnasnexus/pgac312. View

12.

Di Z, Sanyuan S, Hong L, Dahai Y . Enhanced radiosensitivity and G2/M arrest were observed in radioresistant esophageal cancer cells by knocking down RPA expression. Cell Biochem Biophys. 2014; 70(2):887-91. DOI: 10.1007/s12013-014-9995-3. View

13.

Zhou W, Yao Y, Scott A, Wilder-Romans K, Dresser J, Werner C . Purine metabolism regulates DNA repair and therapy resistance in glioblastoma. Nat Commun. 2020; 11(1):3811. PMC: 7393131. DOI: 10.1038/s41467-020-17512-x. View

14.

Zhang Z, Huo H, Liao K, Wang Z, Gong Z, Li Y . RPA1 downregulation enhances nasopharyngeal cancer radiosensitivity via blocking RAD51 to the DNA damage site. Exp Cell Res. 2018; 371(2):330-341. DOI: 10.1016/j.yexcr.2018.08.025. View

15.

Wang R, Shang Y, Chen B, Xu F, Zhang J, Zhang Z . Protein disulfide isomerase blocks the interaction of LC3II-PHB2 and promotes mTOR signaling to regulate autophagy and radio/chemo-sensitivity. Cell Death Dis. 2022; 13(10):851. PMC: 9537141. DOI: 10.1038/s41419-022-05302-w. View

16.

Kim H, Brill S . Rfc4 interacts with Rpa1 and is required for both DNA replication and DNA damage checkpoints in Saccharomyces cerevisiae. Mol Cell Biol. 2001; 21(11):3725-37. PMC: 87010. DOI: 10.1128/MCB.21.11.3725-3737.2001. View

17.

Yam C, Lim H, Surana U . DNA damage checkpoint execution and the rules of its disengagement. Front Cell Dev Biol. 2022; 10:1020643. PMC: 9582513. DOI: 10.3389/fcell.2022.1020643. View

18.

Bhattacharya S, Srinivasan K, Abdisalaam S, Su F, Raj P, Dozmorov I . RAD51 interconnects between DNA replication, DNA repair and immunity. Nucleic Acids Res. 2017; 45(8):4590-4605. PMC: 5416901. DOI: 10.1093/nar/gkx126. View

19.

Gao Y, Chen B, Wang R, Xu A, Wu L, Lu H . Knockdown of RRM1 in tumor cells promotes radio-/chemotherapy induced ferroptosis by regulating p53 ubiquitination and p21-GPX4 signaling axis. Cell Death Discov. 2022; 8(1):343. PMC: 9343379. DOI: 10.1038/s41420-022-01140-z. View

20.

Tian Y, Liu G, Wang H, Tian Z, Cai Z, Zhang F . Valproic acid sensitizes breast cancer cells to hydroxyurea through inhibiting RPA2 hyperphosphorylation-mediated DNA repair pathway. DNA Repair (Amst). 2017; 58:1-12. DOI: 10.1016/j.dnarep.2017.08.002. View