Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells

Beyza Reisoğlu; M. Alper Arslan

doi:10.23902/trkjnat.202573

Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells

Abstract

Background: Triple-negative breast cancer (TNBC) is an aggressive cancer type associated with poor prognosis and limited therapeutic options. Due to the absence of targetable receptors, conventional chemotherapy remains the primary treatment approach. ERCC1 is a critical component of the nucleotide excision repair system, responsible for repairing DNA damaged by platinum-based agents like carboplatin. Heat shock response (HSR) is a fundamental, stressinduced defense mechanism that supports cancer cell survival.

Aims: This study aims to investigate the effects of HSR inhibition by KNK437 on ERCC1 gene expression and carboplatin sensitivity in the TNBC cell line MDA-MB-231. Methods: The IC50 values of carboplatin and KNK437 were determined using WST-8 cytotoxicity assay. ERCC1 gene expression levels were quantified by real-time quantitative polymerase chain reaction. Apoptotic and necrotic cell death induced by carboplatin and KNK437 was assessed by flow cytometry using FITC-Annexin V assay.

Results: The IC50 values of carboplatin and KNK437 were 247.5 µM and 89.74 µM, respectively. Carboplatin or KNK437 monotherapy significantly decreased ERCC1 expression by 42.8% and 49.5%, respectively, while their combined application caused a 54.9% reduction. Furthermore, co-treatment markedly increased total cell death by 34.1% compared to carboplatin alone. Interestingly, necrosis induced by carboplatin shifted toward apoptosis upon co-treatment with KNK437, as confirmed by light microscopy and flow cytometry.

Conclusion: HSR inhibition by KNK437 enhances carboplatin sensitivity in TNBC cells and downregulates ERCC1 gene expression. Given the aggressive nature of TNBC and its limited treatment options, our results suggest that KNK437 may offer therapeutic advantages when combined with carboplatin, particularly in contexts where carboplatin-induced necrosis contributes to inflammationrelated complications.

Keywords

Supporting Institution

Ondokuz Mayıs University Scientific Research Projects Coordination Unit

Project Number

BAP04-A-2024-4886

Ethical Statement

Not applicable.

Thanks

This study is based on the master’s thesis of Beyza Reisoğlu, conducted under the supervision of Assist. Prof. Dr. M. Alper Arslan as the thesis advisor. We would like to thank OMU-KÖKMER Center for their assistance with flow cytometry used in Annexin V cell death assay

References

Akkız, H., Şimşek, H., Balcı, D., Ülger, Y., Onan, E., Akçaer, N., & Delik, A. (2025). Inflammation and cancer: Molecular mechanisms and clinical consequences. Frontiers in Oncology, 15, 1564572. https://doi.org/10.3389/fonc.2025.1564572
Aranda-Anzaldo, A., Dent, M. A. R., Segura-Anaya, E., & MartinezGomez, A. (2024). Protein folding, cellular stress and cancer. Progress in Biophysics and Molecular Biology, 191, 40–57. https://doi.org/10.1016/j.pbiomolbio.2024.07.001
Balvan, J., Krizova, A., Gumulec, J., Raudenska, M., Sladek, Z., Sedlackova, M., Babula, P., Sztalmachova, M., Kizek, R., Chmelik, R., & Masarik, M. (2015). Multimodal holographic microscopy: Distinction between apoptosis and oncosis. PLOS ONE, 10(3), e0121674. https://doi.org/10.1371/journal.pone.0121674
Barrio, S., Gallardo, M., Arenas, A., Ayala, R., Rapado, I., Rueda, D., JimenezUbieto, A., Albizua, E., Burgaleta, C., Gilsanz, F., & Martinez-Lopez, J. (2013). Inhibition of related JAK/STAT pathways with molecular targeted drugs shows strong synergy with ruxolitinib in chronic myeloproliferative neoplasm. British Journal of Haematology, 161(5), 667–676. https://doi.org/10.1111/bjh.12308
Bian, L., Yu, P., Wen, J., Li, N., Huang, W., Xie, X., & Ye, F. (2021). Survival benefit of platinum-based regimen in early stage triple negative breast cancer: A meta-analysis of randomized controlled trials. NPJ Breast Cancer, 7(1), 157. https://doi.org/10.1038/s41523-021-00367-w
Bou Zerdan, M., Ghorayeb, T., Saliba, F., Allam, S., Bou Zerdan, M., Yaghi, M., Bilani, N., Jaafar, R., & Nahleh, Z. (2022). Triple negative breast cancer: Updates on classification and treatment in 2021. Cancers, 14(5), 1253. https://doi.org/10.3390/cancers14051253
Brabec, V., & Kasparkova, J. (2005). Modifications of DNA by platinum complexes: Relation to resistance of tumors to platinum antitumor drugs. Drug Resistance Updates, 8(3), 131–146. https://doi.org/10.1016/j.drup.2005.04.006
Clowse, M. E., & Wigley, F. M. (2003). Digital necrosis related to carboplatin and gemcitabine therapy in systemic sclerosis. The Journal of Rheumatology, 30(6), 1341–1343. https://www.ncbi.nlm.nih.gov/pubmed/12784412

Dai, C., Whitesell, L., Rogers, A. B., & Lindquist, S. (2007). Heat shock factor 1 is a powerful multifaceted modifier of carcinogenesis. Cell, 130(6), 1005–1018. https://doi.org/10.1016/j.cell.2007.07.020
Desai, S., Liu, Z., Yao, J., Patel, N., Chen, J., Wu, Y., Ahn, E. E., Fodstad, O., & Tan, M. (2013). Heat shock factor 1 (HSF1) controls chemoresistance and autophagy through transcriptional regulation of autophagy-related protein 7 (ATG7). Journal of Biological Chemistry, 288(13), 9165–9176. https://doi.org/10.1074/jbc.M112.422071
Grunt, T. W., Somay, C., Pavelka, M., Ellinger, A., Dittrich, E., & Dittrich, C. (1991). The effects of dimethyl sulfoxide and retinoic acid on the cell growth and the phenotype of ovarian cancer cells. Journal of Cell Science, 100(3), 657–666. https://doi.org/10.1242/jcs.100.3.657
Hashmi, A. A., Ajaz, Y., Sajjad, M., Zia, F., Irfan, M., Abu Bakar, S. M., Khan, E. Y., & Faridi, N. (2025). Predictive value of excision repair cross complementation group 1 (ERCC1) by immunohistochemistry for determining neoadjuvant chemotherapy response in triple-negative breast cancers. The Breast Journal, 2025, 8410670. https://doi.org/10.1155/tbj/8410670
Hruban, R. H., Sternberg, S. S., Meyers, P., Fleisher, M., Menendez-Botet, C., & Boitnott, J. K. (1991). Fatal thrombocytopenia and liver failure associated with carboplatin therapy. Cancer Investigation, 9(3), 263–268. https://doi.org/10.3109/07357909109021323
Kawara, H., Akahori, R., Wakasugi, M., Sancar, A., & Matsunaga, T. (2019). DCAF7 is required for maintaining the cellular levels of ERCC1-XPF and nucleotide excision repair. Biochemical and Biophysical Research Communications, 519(1), 204–210. https://doi.org/10.1016/j.bbrc.2019.08.147
Kovács, D., Sigmond, T., Hotzi, B., Bohar, B., Fazekas, D., Deak, V., Vellai, T., & Barna, J. (2019). HSF1Base: A comprehensive database of HSF1 (heat shock factor 1) target genes. International Journal of Molecular Sciences, 20(22), 5815. https://doi.org/10.3390/ijms20225815
Lee, J. (2023). Current treatment landscape for early triple-negative breast cancer (TNBC). Journal of Clinical Medicine, 12(4), 1524. https://doi.org/10.3390/jcm12041524
Li, Q., Yu, J. J., Mu, C., Yunmbam, M. K., Slavsky, D., Cross, C. L., BostickBruton, F., & Reed, E. (2000). Association between the level of ERCC-1 expression and the repair of cisplatin-induced DNA damage in human ovarian cancer cells. Anticancer Research, 20, 645–652.
Lin, C., Cui, J., Peng, Z., Qian, K., Wu, R., Cheng, Y., & Yin, W. (2022). Efficacy of platinum-based and non-platinum-based drugs on triple-negative breast cancer: Meta-analysis. European Journal of Medical Research, 27(1), 201. https://doi.org/10.1186/s40001-022-00839-0
Ma, E. L. B., & El Kashef, W. F. (2017). ERCC1 expression in metastatic triple negative breast cancer patients treated with platinum-based chemotherapy. Asian Pacific Journal of Cancer Prevention, 18(2), 507–513. https://doi.org/10.22034/APJCP.2017.18.2.507
Marteijn, J. A., Lans, H., Vermeulen, W., & Hoeijmakers, J. H. (2014). Understanding nucleotide excision repair and its roles in cancer and ageing. Nature Reviews Molecular Cell Biology, 15(7), 465–481. https://doi.org/10.1038/nrm3822
Mendillo, M. L., Santagata, S., Koeva, M., Bell, G. W., Hu, R., Tamimi, R. M., Fraenkel, E., Ince, T. A., Whitesell, L., & Lindquist, S. (2012). HSF1 drives a transcriptional program distinct from heat shock to support highly malignant human cancers. Cell, 150(3), 549–562. https://doi.org/10.1016/j.cell.2012.06.031
Metzger, R., Leichman, C. G., Danenberg, K. D., Danenberg, P. V., Lenz, H. J., Hayashi, K., Groshen, S., Salonga, D., Cohen, H., Laine, L., Crookes, P., Silberman, H., Baranda, J., Konda, B., & Leichman, L. (1998). ERCC1 mRNA levels complement thymidylate synthase mRNA levels in predicting response and survival for gastric cancer patients receiving combination cisplatin and fluorouracil chemotherapy. Journal of Clinical Oncology, 16(1), 309–316. https://doi.org/10.1200/JCO.1998.16.1.309
Page, T. J., Sikder, D., Yang, L., Pluta, L., Wolfinger, R. D., Kodadek, T., & Thomas, R. S. (2006). Genome-wide analysis of human HSF1 signaling reveals a transcriptional program linked to cellular adaptation and survival. Molecular BioSystems, 2(12), 627–639. https://doi.org/10.1039/b606129j
Pathak, N., Sharma, A., Elavarasi, A., Sankar, J., Deo, S. V. S., Sharma, D. N., Mathur, S., Kumar, S., Prasad, C. P., Kumar, A., & Batra, A. (2022). Moment of truth—Adding carboplatin to neoadjuvant/adjuvant chemotherapy in triple negative breast cancer improves overall survival: An individual participant data and trial-level meta-analysis. The Breast, 64, 7–18. https://doi.org/10.1016/j.breast.2022.04.006
Powers, M. V., & Workman, P. (2007). Inhibitors of the heat shock response: Biology and pharmacology. FEBS Letters, 581(19), 3758–3769. https://doi.org/10.1016/j.febslet.2007.05.040
Sidoni, A., Cartaginese, F., Colozza, M., Gori, S., & Crino, L. (2008). ERCC1 expression in triple negative breast carcinoma: The paradox revisited. Breast Cancer Research and Treatment, 111(3), 569–570. https://doi.org/10.1007/s10549-007-9804-4
de Sousa, G. F., Wlodarczyk, S. R., & Monteiro, G. (2014). Carboplatin: Molecular mechanisms of action associated with chemoresistance. Brazilian Journal of Pharmaceutical Sciences, 50(4), 693–701. https://doi.org/10.1590/S1984-82502014000400004
Tuncer, S., Gurbanov, R., Sheraj, I., Solel, E., Esenturk, O., & Banerjee, S. (2018). Low dose dimethyl sulfoxide driven gross molecular changes have the potential to interfere with various cellular processes. Scientific Reports, 8(1), 14828. https://doi.org/10.1038/s41598-018-33234-z
Verheijen, M., Lienhard, M., Schrooders, Y., Clayton, O., Nudischer, R., Boerno, S., Timmermann, B., Selevsek, N., Schlapbach, R., Gmuender, H., Gotta, S., Geraedts, J., Herwig, R., Kleinjans, J., & Caiment, F. (2019). DMSO induces drastic changes in human cellular processes and epigenetic landscape in vitro. Scientific Reports, 9(1), 4641. https://doi.org/10.1038/s41598-019-40660-0
Voorhees, P. M., Chen, Q., Kuhn, D. J., Small, G. W., Hunsucker, S. A., Strader, J. S., Corringham, R. E., Zaki, M. H., Nemeth, J. A., & Orlowski, R. Z. (2007). Inhibition of interleukin-6 signaling with CNTO 328 enhances the activity of bortezomib in preclinical models of multiple myeloma. Clinical Cancer Research, 13(21), 6469–6478. https://doi.org/10.1158/1078-0432.CCR-07-1293
Vyas, D., Laput, G., & Vyas, A. K. (2014). Chemotherapy-enhanced inflammation may lead to the failure of therapy and metastasis. OncoTargets and Therapy, 7, 1015–1023. https://doi.org/10.2147/OTT.S60114
Wang, S., Zhang, H., Cheng, L., Evans, C., & Pan, C. X. (2010). Analysis of the cytotoxic activity of carboplatin and gemcitabine combination. Anticancer Research, 30(11), 4573–4578. https://www.ncbi.nlm.nih.gov/pubmed/21115908
Welsh, C., Day, R., McGurk, C., Masters, J. R., Wood, R. D., & Koberle, B. (2004). Reduced levels of XPA, ERCC1 and XPF DNA repair proteins in testis tumor cell lines. International Journal of Cancer, 110(3), 352–361. https://doi.org/10.1002/ijc.20134
Yun, C. W., Kim, H. J., Lim, J. H., & Lee, S. H. (2019). Heat shock proteins: Agents of cancer development and therapeutic targets in anti-cancer therapy. Cells, 9(1), 60. https://doi.org/10.3390/cells9010060

Details

Primary Language

English

Subjects

Cell Metabolism

Journal Section

Research Article

Authors

Beyza Reisoğlu
Türkiye

M. Alper Arslan
Türkiye

Publication Date

July 9, 2026

Submission Date

August 18, 2025

Acceptance Date

December 9, 2025

Published in Issue

Year 2026 Volume: 27 Number: 1

DOI

https://doi.org/10.23902/trkjnat.202573

IZ

https://izlik.org/JA58NF72JM

Cite

RIS / Bibtex

APA

Reisoğlu, B., & Arslan, M. A. (2026). Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells. Trakya University Journal of Natural Sciences, 27(1), 47-56. https://doi.org/10.23902/trkjnat.202573

AMA

1.Reisoğlu B, Arslan MA. Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells. Trakya Univ J Nat Sci. 2026;27(1):47-56. doi:10.23902/trkjnat.202573

Chicago

Reisoğlu, Beyza, and M. Alper Arslan. 2026. “Chemical Inhibition of the Heat Shock Response Downregulates ERCC1 Gene Expression and Shifts Carboplatin-Induced Necrosis to Apoptosis in TNBC Cells”. Trakya University Journal of Natural Sciences 27 (1): 47-56. https://doi.org/10.23902/trkjnat.202573.

EndNote

Reisoğlu B, Arslan MA (July 1, 2026) Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells. Trakya University Journal of Natural Sciences 27 1 47–56.

IEEE

[1]B. Reisoğlu and M. A. Arslan, “Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells”, Trakya Univ J Nat Sci, vol. 27, no. 1, pp. 47–56, July 2026, doi: 10.23902/trkjnat.202573.

ISNAD

Reisoğlu, Beyza - Arslan, M. Alper. “Chemical Inhibition of the Heat Shock Response Downregulates ERCC1 Gene Expression and Shifts Carboplatin-Induced Necrosis to Apoptosis in TNBC Cells”. Trakya University Journal of Natural Sciences 27/1 (July 1, 2026): 47-56. https://doi.org/10.23902/trkjnat.202573.

JAMA

1.Reisoğlu B, Arslan MA. Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells. Trakya Univ J Nat Sci. 2026;27:47–56.

MLA

Reisoğlu, Beyza, and M. Alper Arslan. “Chemical Inhibition of the Heat Shock Response Downregulates ERCC1 Gene Expression and Shifts Carboplatin-Induced Necrosis to Apoptosis in TNBC Cells”. Trakya University Journal of Natural Sciences, vol. 27, no. 1, July 2026, pp. 47-56, doi:10.23902/trkjnat.202573.

Vancouver

1.Beyza Reisoğlu, M. Alper Arslan. Chemical inhibition of the heat shock response downregulates ERCC1 gene expression and shifts carboplatin-induced necrosis to apoptosis in TNBC cells. Trakya Univ J Nat Sci. 2026 Jul. 1;27(1):47-56. doi:10.23902/trkjnat.202573