Inhibition of Cyclin F Promotes Cellular Senescence through Cyclin-dependent Kinase 1-mediated Cell Cycle Regulation

Abstract

Objective

Kidney renal clear cell carcinoma (KIRC) is a common renal malignancy that has a poor prognosis. As a member of the F box family, cyclin F (CCNF) plays an important regulatory role in normal tissues and tumors. However, the underlying mechanism by which CCNF promotes KIRC proliferation still remains unclear.

Methods

Bioinformatics methods were used to analyze The Cancer Genome Atlas (TCGA) database to obtain gene expression and clinical prognosis data. The CCK8 assay, EdU assay, and xenograft assay were used to detect cell proliferation. The cell senescence and potential mechanism were assessed by SA-β-gal staining, Western blotting, as well as ELISA.

Results

Our data showed that CCNF was highly expressed in KIRC patients. Meanwhile, downregulation of CCNF inhibited cell proliferation in vivo and in vitro. Further studies showed that the reduction of CCNF promoted cell senescence by decreasing cyclin-dependent kinase 1 (CDK1), increasing the proinflammatory factors interleukin (IL)-6 and IL-8, and then enhancing the expression of p21 and p53.

Conclusion

We propose that the high expression of CCNF in KIRC may play a key role in tumorigenesis by regulating cell senescence. Therefore, CCNF shows promise as a new biomarker to predict the clinical prognosis of KIRC patients and as an effective therapeutic target.

Keywords

cyclin F / kidney renal clear cell carcinoma / clinical outcome / cyclin-dependent kinase 1 / senescence

References

[1] PadalaSA, BarsoukA, ThandraKC, et al.. Epidemiology of Renal Cell Carcinoma. World J Oncol, 2022, 11(3): 79-87

[2] HsiehJJ, PurdueMP, SignorettiS, et al.. Renal Cell Carcinoma. Nat Rev Dis Primers, 2017, 3: 17009

[3] MillerKD, NogueiraL, MariottoAB, et al.. Cancer Treatment and Survivorship Statistics, 2019. CA Cancer J Clin, 2019, 69(5): 363-385

[4] MotzerRJ, JonaschE, AgarwalN, et al.. Kidney Cancer, Version 3.2015. J Natl Compr Canc Netw, 2015, 13(2): 151-159

[5] D’AngiolellaV, DonatoV, VijayakumarS, et al.. Scf(Cyclin F) Controls Centrosome Homeostasis and Mitotic Fidelity through Cp110 Degradation. Nature, 2010, 466(7302): 138-142

[6] YangZL, YangH, TuoT, et al.. Increased Expression of Cyclin F in Liver Cancer Predicts Poor Prognosis: A Study Based on Tcga Database. Medicine (Baltimore), 2021, 100(31): e26623-e23

[7] D’AngiolellaV, DonatoV, ForresterFM, et al.. Cyclin F-Mediated Degradation of Ribonucleotide Reductase M2 Controls Genome Integrity and DNA Repair. Cell, 2012, 149(5): 1023-1034

[8] GalperJ, RaynerSL, HoganAL, et al.. Cyclin F: A Component of an E3 Ubiquitin Ligase Complex with Roles in Neurodegeneration and Cancer. Int J Biochem Cell Biol, 2017, 89: 216-220

[9] ChangSC, HungCS, ZhangBX, et al.. A Novel Signature of Ccnf-Associated E3 Ligases Collaborate and Counter Each Other in Breast Cancer. Cancers (Basel), 2021, 13(12): 2873

[10] FuJ, QiuH, CaiM, et al.. Low Cyclin F Expression in Hepatocellular Carcinoma Associates with Poor Differentiation and Unfavorable Prognosis. Cancer Sci, 2013, 104(4): 508-515

[11] XiaS, LinY, LinJ, et al.. Increased Expression of Ticrr Predicts Poor Clinical Outcomes: A Potential Therapeutic Target for Papillary Renal Cell Carcinoma. Front Genet, 2020, 11: 605378

[12] SubramanianA, TamayoP, MoothaVK, et al.. Gene Set Enrichment Analysis: A Knowledge-Based Approach for Interpreting Genome-Wide Expression Profiles. Proc Natl Acad Sci USA, 2005, 102(43): 15545-15550

[13] YuG, WangLG, HanY, et al.. Clusterprofiler: An R Package for Comparing Biological Themes among Gene Clusters. OMICS, 2012, 16(5): 284-287

[14] SzklarczykD, GableAL, LyonD, et al.. String V11: Protein-Protein Association Networks with Increased Coverage, Supporting Functional Discovery in Genome-Wide Experimental Datasets. Nucleic Acids Res, 2019, 47(D1): D607-D613

[15] LiuJ, LichtenbergT, HoadleyKA, et al.. An Integrated Tcga Pan-Cancer Clinical Data Resource to Drive High-Quality Survival Outcome Analytics. Cell, 2018, 173(2): 400-416.e11

[16] BaiC, RichmanR, ElledgeSJ. Human Cyclin F. EMBO J, 1994, 13(24): 6087-6098

[17] LiY, GuoH, WangZ, et al.. Cyclin F and Kif20a, Foxm1 Target Genes, Increase Proliferation and Invasion of Ovarian Cancer Cells. Exp Cell Res, 2020, 395(2): 112212

[18] ComissoE, ScarolaM, RossoM, et al.. Oct4 Controls Mitotic Stability and Inactivates the Rb Tumor Suppressor Pathway to Enhance Ovarian Cancer Aggressiveness. Oncogene, 2017, 36(30): 4253-4266

[19] LiaoH, JiF, YingS. Cdk1: Beyond Cell Cycle Regulation. Aging (Albany NY), 2017, 9(12): 2465-2466

[20] BaeT, WeonKY, LeeJW, et al.. Restoration of Paclitaxel Resistance by Cdk1 Intervention in Drug-Resistant Ovarian Cancer. Carcinogenesis, 2015, 36(12): 1561-1571

[21] MüllersE, Silva CascalesH, BurdovaK, et al.. Residual Cdk1/2 Activity after DNA Damage Promotes Senescence. Aging Cell, 2017, 16(3): 575-584

[22] RiesslandM, KolisnykB, KimTW, et al.. Loss of Satb1 Induces P21-Dependent Cellular Senescence in Post-Mitotic Dopaminergic Neurons. Cell Stem Cell, 2019, 25(4): 514–530-e8