Abstract
Cell division cyclin 25 C (CDC25C) functions as an antigen linked to hepatocellular carcinoma (HCC) and is vital for its diagnosis, treatment, and prognosis. However, the precise pathways underlying CDC25C-mediated inhibition of HCC growth remain incompletely understood. In this study, we created a CDC25C-downregulation model in AML12 and Hepa1-6 cell lines, complemented by a tumor xenograft in C57BL/6 mice. We evaluated the malignant biological behaviors and subcellular structural morphology of the CDC25C-downregulation model. Additionally, we quantified mitochondrial calcium levels, reactive oxygen species (ROS) concentrations, mitochondrial stress-related molecules, and autophagy-related proteins. Furthermore, we observed the morphological characteristics of apoptosis in CDC25C-downregulated cells, evaluated the apoptosis rate, and identified key molecules involved in the mitochondrial apoptosis pathway. Our findings indicated that the downregulation of CDC25C inhibited the proliferation, migration and invasion of HCC cells. Further analysis demonstrated that apoptosis in HCC cells was ultimately induced by CDC25C downregulation, which also triggered the mitochondrial stress response and autophagy. In contrast, normal hepatocytes exhibited the opposite effect, with the exception of autophagy. In summary, downregulation of CDC25C does not inhibit the growth of normal hepatocytes but inhibits HCC progression; its anti-HCC effect may be associated with the mitochondrial stress response as well as mitochondria-mediated autophagy and apoptosis.
Data availability
No datasets were generated or analysed during the current study.
References
Tong, H. et al. MACC1-AS1 promotes hepatocellular carcinoma cell invasion and proliferation by regulating PAX8. Aging (Albany NY). 12, 70–79. https://doi.org/10.18632/aging.102585 (2020).
Wong, G. L. et al. Novel machine learning models outperform risk scores in predicting hepatocellular carcinoma in patients with chronic viral hepatitis. JHEP Rep. 4, 100441. https://doi.org/10.1016/j.jhepr.2022.100441 (2022).
Yu, Z. et al. G250 Antigen-Targeting Drug-Loaded nanobubbles combined with ultrasound targeted nanobubble destruction: A potential novel treatment for renal cell carcinoma. Int. J. Nanomed. 15, 81–95. https://doi.org/10.2147/IJN.S230879 (2020).
Liu, K. et al. The role of CDC25C in cell cycle regulation and clinical cancer therapy: a systematic review. Cancer Cell. Int. 20, 213. https://doi.org/10.1186/s12935-020-01304-w (2020).
Liu, J. S. et al. Aloperine induces apoptosis and G2/M cell cycle arrest in hepatocellular carcinoma cells through the PI3K/Akt signaling pathway. Phytomedicine 61, 152843. https://doi.org/10.1016/j.phymed.2019.152843 (2019).
Wang, H. K., Xu, X. H., Wang, S. M. & Zhang, H. Y. Preparation of hepatocellular carcinoma mRNA vaccines based on potential tumor targets and immunophenotypes. Transl Cancer Res. 13, 173–190. https://doi.org/10.21037/tcr-23-1237 (2024).
Zhu, S. et al. hsa_circ_0013401 accelerates the growth and metastasis and prevents apoptosis and autophagy of neuroblastoma cells by sponging miR-195 to release PAK2. Oxid. Med. Cell. Longev. 2021, 9936154. https://doi.org/10.1155/2021/9936154 (2021).
Hu, J. et al. Alanyl-Glutamine protects against Lipopolysaccharide-Induced liver injury in mice via alleviating oxidative Stress, inhibiting Inflammation, and regulating autophagy. Antioxid. (Basel). 11, 1070. https://doi.org/10.3390/antiox11061070 (2022).
Zheng, Z. et al. Site-1 protease controls osteoclastogenesis by mediating LC3 transcription. Cell. Death Differ. 28, 2001–2018. https://doi.org/10.1038/s41418-020-00731-6 (2021).
Liu, Y. et al. Phosphoinositide-3-kinase regulatory subunit 4 participates in the occurrence and development of amyotrophic lateral sclerosis by regulating autophagy. Neural Regen Res. 17, 1609–1616. https://doi.org/10.4103/1673-5374.330621 (2022).
Tran, S., Fairlie, W. D. & Lee, E. F. BECLIN1: protein Structure, function and regulation. Cells 10, 1522. https://doi.org/10.3390/cells10061522 (2021).
Uzhachenko, R., Shanker, A., Yarbrough, W. G. & Ivanova, A. V. Mitochondria, calcium, and tumor suppressor Fus1: at the crossroad of cancer, inflammation, and autoimmunity. Oncotarget 6, 20754–20772. https://doi.org/10.18632/oncotarget.4537 (2015).
Lee, Y. G., Park, D. H. & Chae, Y. C. Role of mitochondrial stress response in cancer progression. Cells 11, 771. https://doi.org/10.3390/cells11050771 (2022).
Liu, Y. et al. ATF5 regulates tubulointerstitial injury in diabetic kidney disease via mitochondrial unfolded protein response. Mol. Med. 29, 57. https://doi.org/10.1186/s10020-023-00651-4 (2023).
Kumar, R. et al. A mitochondrial unfolded protein response inhibitor suppresses prostate cancer growth in mice via HSP60. J. Clin. Invest. 132, e149906. https://doi.org/10.1172/JCI149906 (2022).
Lopez-Crisosto, C. et al. Endoplasmic reticulum-mitochondria coupling increases during doxycycline-induced mitochondrial stress in HeLa cells. Cell. Death Dis. 12, 657. https://doi.org/10.1038/s41419-021-03945-9 (2021).
Kang, L., Tian, Y., Guo, X., Chu, X. & Xue, Y. Long noncoding RNA ANPODRT overexpression protects nucleus pulposus cells from oxidative stress and apoptosis by activating Keap1-Nrf2 signaling. Oxid. Med. Cell Longev. 2021 6645005. https://doi.org/10.1155/2021/6645005 (2021).
Liao, K. F., Chiu, T. L., Chang, S. F., Wang, M. J. & Chiu, S. C. Hispolon induces Apoptosis, suppresses migration and invasion of glioblastoma cells and inhibits GBM xenograft tumor growth in vivo. Molecules 26, 4497. https://doi.org/10.3390/molecules26154497 (2021).
Sun, L. et al. Beclin-1-independent autophagy mediates programmed cancer cell death through interplays with Endoplasmic reticulum and/or mitochondria in Colbat chloride-induced hypoxia. Am. J. Cancer Res. 5, 2626–2642 (2015). PMID: 26609472.
Li, Y. F. et al. Cell division Cyclin 25 C knockdown inhibits hepatocellular carcinoma development by inducing Endoplasmic reticulum stress. World J. Gastroenterol. 30, 2564–2574. https://doi.org/10.3748/wjg.v30.i19.2564 (2024).
Xun, R., Lu, H. & Wang, X. Identification of CDC25C as a potential biomarker in hepatocellular carcinoma using bioinformatics analysis. Technol. Cancer Res. Treat. 19, 1533033820967474. https://doi.org/10.1177/1533033820967474 (2020).
Chan, M. L. et al. Zerumbone, a ginger sesquiterpene, induces apoptosis and autophagy in human hormone-refractory prostate cancers through tubulin binding and crosstalk between Endoplasmic reticulum stress and mitochondrial insult. Naunyn Schmiedebergs Arch. Pharmacol. 388, 1223–1236. https://doi.org/10.1007/s00210-015-1152-z (2015).
Zhang, S. et al. Homocysteine promotes atherosclerosis through macrophage pyroptosis via Endoplasmic reticulum stress and calcium disorder. Mol. Med. 29, 73. https://doi.org/10.1186/s10020-023-00656-z (2023).
Joseph, L. C. et al. PKCδ causes sepsis-induced cardiomyopathy by inducing mitochondrial dysfunction. Am. J. Physiol. Heart Circ. Physiol. 318, H778–H786. https://doi.org/10.1152/ajpheart.00749.2019 (2020).
Belli, M. et al. Oxygen concentration alters mitochondrial structure and function in in vitro fertilized preimplantation mouse embryos. Hum. Reprod. 35, 1476. https://doi.org/10.1093/humrep/deaa047 (2020).
Horibe, T. & Hoogenraad, N. J. The Chop gene contains an element for the positive regulation of the mitochondrial unfolded protein response. PLoS One. 2, e835. https://doi.org/10.1371/journal.pone.0000835 (2007).
Wardelmann, K. et al. Insulin action in the brain regulates mitochondrial stress responses and reduces diet-induced weight gain. Mol. Metab. 21, 68–81. https://doi.org/10.1016/j.molmet.2019.01.001 (2019).
Dancourt, J. & Melia, T. J. Lipidation of the autophagy proteins LC3 and GABARAP is a membrane-curvature dependent process. Autophagy 10, 1470–1471. https://doi.org/10.4161/auto.29468 (2014).
Lee, Y. & Weihl, C. C. Regulation of SQSTM1/p62 via UBA domain ubiquitination and its role in disease. Autophagy 13, 1615–1616. https://doi.org/10.1080/15548627.2017.1339845 (2017).
Li, X. et al. Beclin1- and Atg13-dependent autophagy activation and Morroniside have synergistic effect on osteoblastogenesis. Exp. Biol. Med. (Maywood). 247, 1764–1775. https://doi.org/10.1177/15353702221116879 (2022).
Wirawan, E. et al. Caspase-mediated cleavage of Beclin-1 inactivates Beclin-1-induced autophagy and enhances apoptosis by promoting the release of proapoptotic factors from mitochondria. Cell. Death Dis. 1, e18. https://doi.org/10.1038/cddis.2009.16 (2010).
Saha, A., Saleem, S., Paidi, R. K. & Biswas, S. C. BH3-only proteins Puma and Beclin1 regulate autophagic death in neurons in response to Amyloid-β. Cell. Death Discov. 7, 356. https://doi.org/10.1038/s41420-021-00748-x (2021).
Zeng, S. et al. Role of GABAA receptor depolarization-mediated VGCC activation in sevoflurane-induced cognitive impairment in neonatal mice. Front. Cell. Neurosci. 16, 964227. https://doi.org/10.3389/fncel.2022.964227 (2022).
Liu, Z. et al. ZWZ-3, a fluorescent probe targeting mitochondria for melanoma imaging and therapy. Front. Pharmacol. 13, 829684. https://doi.org/10.3389/fphar.2022.829684 (2022).
Deng, X. et al. Paeoniflorin protects hepatocytes from APAP-induced damage through launching autophagy via the MAPK/mTOR signaling pathway. Cell. Mol. Biol. Lett. 29, 119. https://doi.org/10.1186/s11658-024-00631-4 (2024).
Funding
This work was supported by the Natural Science Foundation of Guangxi (2023GXNSFAA026070 and 2025GXNSFAA069047) and the Middle/Young aged Teachers’ Research Ability Improvement Project of Guangxi Higher Education (2024KY0107).
Author information
Authors and Affiliations
Contributions
Farong Mo and Naixia Chao designed and supervised the study. Xinyu Miao, Fangyuan Zheng, Xiaohui Mo, Lifei Wang, Zhengziyi Zhang, Yanfei Li and Yaoyao Zhang performed experiments. Xinyu Miao and Fangyuan Zheng wrote the original manuscript and produced figures. Critical revisions were made by Farong Mo and Naixia Chao. All authors have read and agreed to the published version of the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Institutional review board
All animal experiments were reviewed and approved by the Ethics Review Committee for Animal Experiments at Guangxi Medical University (Approval Number: 202201093) and followed the ARRIVE guidelines. We confirm that all experiments were performed in accordance with relevant guidelines and regulations.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Miao, X., Zheng, F., Mo, X. et al. CDC25C downregulation suppresses HCC growth via mitochondrial stress-induced autophagy and apoptosis. Sci Rep (2026). https://doi.org/10.1038/s41598-026-36351-2
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-36351-2
