Targeting the Anaphase-promoting Complex/cyclosome (APC/C) Enhanced Antiproliferative and Apoptotic Response in Bladder Cancer

Overview

Journal Saudi J Biol Sci

Specialty Biology

Date 2023 Feb 16

PMID 36794046

Authors

Hatice Sevim Nalkiran

Dilara Akcora Yildiz

Faruk Saydam

Ali Irfan Guzel

Ihsan Nalkiran

Affiliations

Soon will be listed here.

Abstract

Improving the chemotherapy sensitivity of bladder cancer is a current clinical challenge. It is critical to seek out effective combination therapies that include low doses of cisplatin due to its dose-limiting toxicity. This study aims to investigate the cytotoxic effects of the combination therapy including proTAME, a small molecule inhibitor, targeting Cdc-20 and to determine the expression levels of several APC/C pathway-related genes that may play a role in the chemotherapy response of RT-4 (bladder cancer) and ARPE-19 (normal epithelial) cells. The IC20 and IC50 values were determined by MTS assay. The expression levels of apoptosis-associated ( and and APC/C-associated , , and genes were assessed by qRT-PCR. Cell colonization ability and apoptosis were examined by clonogenic survival experiment and Annexin V/PI staining, respectively. Low-dose combination therapy showed a superior inhibition effect on RT-4 cells by increasing cell death and inhibiting colony formation. Triple-agent combination therapy further increased the percentage of late apoptotic and necrotic cells compared to the doublet-therapy with gemcitabine and cisplatin. ProTAME-containing combination therapies resulted in an elevation in Bax/Bcl-2 ratio in RT-4 cells, while a significant decrease was observed in proTAME-treated ARPE-19 cells. Cdc-20 expression in proTAME combined treatment groups were found to be decreased compared to their control groups. Low-dose triple-agent combination induced cytotoxicity and apoptosis in RT-4 cells effectively. It is essential to evaluate the role of APC/C pathway-associated potential biomarkers as therapeutic targets and define new combination therapy regimens to achieve improved tolerability in bladder cancer patients in the future.

Citing Articles

CDC20 and CCNB1 Overexpression as Prognostic Markers in Bladder Cancer.

Sevim Nalkiran H, Biri I, Nalkiran I, Uzun H, Durur S, Bedir R Diagnostics (Basel). 2025; 15(1.

PMID: 39795587 PMC: 11719780. DOI: 10.3390/diagnostics15010059.

Bioinformatics analysis and experimental validation reveal that CDC20 overexpression promotes bladder cancer progression and potential underlying mechanisms.

Liu Y, Zou S, Gao X Genes Genomics. 2024; 46(4):437-449.

PMID: 38438666 DOI: 10.1007/s13258-024-01505-x.

References

Spiess P, Agarwal N, Bangs R, Boorjian S, Buyyounouski M, Clark P . Bladder Cancer, Version 5.2017, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw. 2017; 15(10):1240-1267. DOI: 10.6004/jnccn.2017.0156. View

Sonpavde G, Watson D, Tourtellott M, Cowey C, Hellerstedt B, Hutson T . Administration of cisplatin-based chemotherapy for advanced urothelial carcinoma in the community. Clin Genitourin Cancer. 2012; 10(1):1-5. DOI: 10.1016/j.clgc.2011.11.005. View

Kato T, Daigo Y, Aragaki M, Ishikawa K, Sato M, Kaji M . Overexpression of CDC20 predicts poor prognosis in primary non-small cell lung cancer patients. J Surg Oncol. 2012; 106(4):423-30. DOI: 10.1002/jso.23109. View

Flaig T, Spiess P, Agarwal N, Bangs R, Boorjian S, Buyyounouski M . Bladder Cancer, Version 3.2020, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw. 2020; 18(3):329-354. DOI: 10.6004/jnccn.2020.0011. View

Galsky M, Pal S, Chowdhury S, Harshman L, Crabb S, Wong Y . Comparative effectiveness of gemcitabine plus cisplatin versus methotrexate, vinblastine, doxorubicin, plus cisplatin as neoadjuvant therapy for muscle-invasive bladder cancer. Cancer. 2015; 121(15):2586-93. DOI: 10.1002/cncr.29387. View

Rizzo M, Giannatempo P, Porta C . Biological Therapeutic Advances for the Treatment of Advanced Urothelial Cancers. Biologics. 2021; 15:441-450. PMC: 8570720. DOI: 10.2147/BTT.S290311. View

Rouette A, Parent S, Girouard J, Leblanc V, Asselin E . Cisplatin increases B-cell-lymphoma-2 expression via activation of protein kinase C and Akt2 in endometrial cancer cells. Int J Cancer. 2011; 130(8):1755-67. DOI: 10.1002/ijc.26183. View

Zeng X, King R . An APC/C inhibitor stabilizes cyclin B1 by prematurely terminating ubiquitination. Nat Chem Biol. 2012; 8(4):383-92. PMC: 3307893. DOI: 10.1038/nchembio.801. View

Lee S, Kim J, Nam H, Gao B, Yin P, Qin B . Parkin Regulates Mitosis and Genomic Stability through Cdc20/Cdh1. Mol Cell. 2015; 60(1):21-34. PMC: 4592523. DOI: 10.1016/j.molcel.2015.08.011. View

10.

Wan L, Chen M, Cao J, Dai X, Yin Q, Zhang J . The APC/C E3 Ligase Complex Activator FZR1 Restricts BRAF Oncogenic Function. Cancer Discov. 2017; 7(4):424-441. PMC: 5380472. DOI: 10.1158/2159-8290.CD-16-0647. View

11.

Bhattacharya S, Ray R, Chaum E, Johnson D, Johnson L . Inhibition of Mdm2 sensitizes human retinal pigment epithelial cells to apoptosis. Invest Ophthalmol Vis Sci. 2011; 52(6):3368-80. PMC: 3109032. DOI: 10.1167/iovs.10-6991. View

12.

Eguren M, Alvarez-Fernandez M, Garcia F, Lopez-Contreras A, Fujimitsu K, Yaguchi H . A synthetic lethal interaction between APC/C and topoisomerase poisons uncovered by proteomic screens. Cell Rep. 2014; 6(4):670-83. DOI: 10.1016/j.celrep.2014.01.017. View

13.

Zeng X, Sigoillot F, Gaur S, Choi S, Pfaff K, Oh D . Pharmacologic inhibition of the anaphase-promoting complex induces a spindle checkpoint-dependent mitotic arrest in the absence of spindle damage. Cancer Cell. 2010; 18(4):382-95. PMC: 2957475. DOI: 10.1016/j.ccr.2010.08.010. View

14.

Kim S, Yu H . Mutual regulation between the spindle checkpoint and APC/C. Semin Cell Dev Biol. 2011; 22(6):551-8. PMC: 3225258. DOI: 10.1016/j.semcdb.2011.03.008. View

15.

Mondal G, Sengupta S, Panda C, Gollin S, Saunders W, Roychoudhury S . Overexpression of Cdc20 leads to impairment of the spindle assembly checkpoint and aneuploidization in oral cancer. Carcinogenesis. 2006; 28(1):81-92. DOI: 10.1093/carcin/bgl100. View

16.

Rape M, Reddy S, Kirschner M . The processivity of multiubiquitination by the APC determines the order of substrate degradation. Cell. 2006; 124(1):89-103. DOI: 10.1016/j.cell.2005.10.032. View

17.

Roufayel R, Younes K, Al-Sabi A, Murshid N . BH3-Only Proteins Noxa and Puma Are Key Regulators of Induced Apoptosis. Life (Basel). 2022; 12(2). PMC: 8875537. DOI: 10.3390/life12020256. View

18.

Zaravinos A, Lambrou G, Boulalas I, Delakas D, Spandidos D . Identification of common differentially expressed genes in urinary bladder cancer. PLoS One. 2011; 6(4):e18135. PMC: 3070717. DOI: 10.1371/journal.pone.0018135. View

19.

Silva G, Filoni L, Salvadori M, Salvadori D . Gemcitabine/Cisplatin Treatment Induces Concomitant SERTAD1, CDKN2B and GADD45A Modulation and Cellular Changes in Bladder Cancer Cells Regardless of the Site of TP53 Mutation. Pathol Oncol Res. 2017; 24(2):407-417. DOI: 10.1007/s12253-017-0255-x. View

20.

Izumi K, Iwamoto H, Yaegashi H, Shigehara K, Nohara T, Kadono Y . Gemcitabine Plus Cisplatin Split Gemcitabine Plus Carboplatin for Advanced Urothelial Cancer With Cisplatin-unfit Renal Function. In Vivo. 2018; 33(1):167-172. PMC: 6364082. DOI: 10.21873/invivo.11454. View