The evaluation of the HSP70 and TCP1 status of circulating tumor cells derived from metastatic colorectal cancer patients

Year 2025, Volume: 42 Issue: 4, 417 - 422, 31.12.2025

Dilek Pirim , Berkcan Dogan , Fatih Atilla Bağcı , Özgen Işık , Türkkan Evrensel

Abstract

Circulating tumor cells (CTCs) are novel biomarkers for liquid biopsy, emerging as a promising research area for colorectal cancer (CRC) management. However, the heterogeneity of CTC phenotypes poses a significant challenge for their reliable detection in blood. The heat shock protein 70 (HSP70) and chaperonin-containing TCP1 (TCP1) have crucial roles in cancer pathogenesis, but their clinical value for CTC characterization remains unknown. We assessed the expression status of the HSP70 and TCP1 in CRC-derived CTCs to evaluate their biomarker potential for CTC identification. Peripheral blood samples were collected from metastatic CRC (mCRC) patients. The enrichments of CTCs were performed by the AdnaTest protocol, and the expression status of the HSP70 and TCP1 was analyzed using PCR-based analysis. Additionally, we analyzed the correlations between these genes and CRC-specific markers (CEA, EGFR, and EpCAM) along with clinicopathological characteristics. HSP70 positivity was observed in 55.32% of mCRC patients, and TCP1 positivity was detected in only 6 samples. Nine samples that were negative for CRC-specific markers showed expressions of HSP70 and TCP1. Notably, no TCP1 positivity was seen in individuals who received more than five systematic treatments. Overall, this study highlights the importance of expanding CTC biomarker panels beyond conventional markers to capture the full spectrum of tumor cell diversity in circulation. Integrating novel molecular markers such as HSP70 and TCP1 may improve the sensitivity of CTC-based diagnostics and facilitate personalized monitoring of disease progression and treatment response in CRC patients.

Keywords

Circulating tumor cells , metastatic colorectal cancer , HSP70 , TCP1 , liquid biopsy

Ethical Statement

All protocols were approved by the Institutional Review Board (IRB) of the Bursa Uludag University (Approval date: 14.02.2023, Approval no: 2023-3/39) and conducted following the relevant ethical guidelines/regulations as specified by the Declaration of Helsinki. Each participant gave informed consent before sample collection.

Supporting Institution

Bursa Uludag University

Project Number

This research was funded by Bursa Uludag University Scientific Research Unit, Türkiye (Grant No: FHIZ-2023-1466 and FOA-2021-625).

References

• Morgan E, Arnold M, Gini A, Lorenzoni V, Cabasag CJ, Laversanne M, et al. Global burden of colorectal cancer in 2020 and 2040: incidence and mortality estimates from GLOBOCAN. Gut. 2023 Feb 1;72(2):338–44.
• Biller LH, Schrag D. Diagnosis and Treatment of Metastatic Colorectal Cancer: A Review. JAMA. 2021 Feb 16;325(7):669–85.
• Kiran NS, Yashaswini C, Maheshwari R, Bhattacharya S, Prajapati BG. Advances in Precision Medicine Approaches for Colorectal Cancer: From Molecular Profiling to Targeted Therapies. ACS Pharmacol Transl Sci. 2024 Apr 12;7(4):967–90.
• Zheng Z, Yu T, Zhao X, Gao X, Zhao Y, Liu G. Intratumor heterogeneity: A new perspective on colorectal cancer research. Cancer Med. 2020;9(20):7637–45.
• Delcuratolo MD, Modrego-Sánchez A, Bungaro M, Antón-Pascual B, Teran S, Dipace V, et al. Liquid Biopsy in Advanced Colorectal Cancer: Clinical Applications of Different Analytes. J Mol Pathol (Basel). 2023 Sep;4(3):128– 55.
• Prathibha K, Swaroop G. Isolation and enumeration of Circulating Tumor Cells (CTC) as prognostic and predictive biomarkers in Post-Operative m-CRC. Clin Investig (Lond). 2021 May;11(3).
• Petrik J, Verbanac D, Fabijanec M, Hulina-Tomašković A, Čeri A, Somborac-Bačura A, et al. Circulating Tumor Cells in Colorectal Cancer: Detection Systems and Clinical Utility. Int J Mol Sci. 2022 Jan;23(21):13582.
• Ju S, Chen C, Zhang J, Xu L, Zhang X, Li Z, et al. Detection of circulating tumor cells: opportunities and challenges. Biomark Res. 2022 Aug 13;10(1):58.
• Debnath J, Gammoh N, Ryan KM. Autophagy and autophagy-related pathways in cancer. Nat Rev Mol Cell Biol. 2023 Aug;24(8):560–75.
• Assaye MA, Gizaw ST. Chaperone-Mediated Autophagy and Its Implications for Neurodegeneration and Cancer. IJGM. 2022 Jun 15;15:5635–49.
• Hartl FU, Bracher A, Hayer-Hartl M. Molecular chaperones in protein folding and proteostasis. Nature. 2011 Jul 20;475(7356):324–32.
• Conway de Macario E, Pitruzzella A, D’Amico AG. Role of Molecular Chaperones in Carcinogenesis: Mechanism, Diagnosis, and Treatment. J Oncol. 2020 Apr 20;2020:7437629.
• Calderwood SK, Murshid A. Molecular Chaperone Accumulation in Cancer and Decrease in Alzheimer’s Disease: The Potential Roles of HSF1. Front Neurosci. 2017;11:192.
• Zhang Z, Jing J, Ye Y, Chen Z, Jing Y, Li S, et al. Characterization of the dual functional effects of heat shock proteins (HSPs) in cancer hallmarks to aid development of HSP inhibitors. Genome Med. 2020 Nov 23;12(1):101.
• Santagata S, Hu R, Lin NU, Mendillo ML, Collins LC, Hankinson SE, et al. High levels of nuclear heat-shock factor 1 (HSF1) are associated with poor prognosis in breast cancer. Proc Natl Acad Sci U S A. 2011 Nov 8;108(45):18378–83.
• Chanteloup G, Cordonnier M, Isambert N, Bertaut A, Marcion G, Garrido C, et al. Membrane-bound exosomal HSP70 as a biomarker for detection and monitoring of malignant solid tumours: a pilot study. Pilot Feasibility Stud. 2020 Mar 3;6:35.
• El-Sheikh NM, Abulsoud AI, Fawzy A, Wasfey EF, Hamdy NM. LncRNA NNT-AS1/hsa-miR-485-5p/HSP90 axis in-silico and clinical prospect correlated-to histologic grades-based CRC stratification: A step toward ncRNA Precision. Pathol Res Pract. 2023 Jul;247:154570.
• Sha G, Jiang Z, Zhang W, Jiang C, Wang D, Tang D. The multifunction of HSP70 in cancer: Guardian or traitor to the survival of tumor cells and the next potential therapeutic target. Int Immunopharmacol. 2023 Sep;122:110492
• Moradi-Marjaneh R, Paseban M, Moradi Marjaneh M. Hsp70 inhibitors: Implications for the treatment of colorectal cancer. IUBMB Life. 2019;71(12):1834–45.
• Ghozlan H, Cox A, Nierenberg D, King S, Khaled AR. The TRiCky Business of Protein Folding in Health and Disease. Front Cell Dev Biol. 2022;10:906530.
• Vallin J, Grantham J. The role of the molecular chaperone CCT in protein folding and mediation of cytoskeleton- associated processes: implications for cancer cell biology. Cell Stress Chaperones. 2019 Jan;24(1):17–27.
• Cheng L, Chen X, Zhang L, Qin N, An J, Zhu J, et al. A potential tumor marker: Chaperonin containing TCP‑1 controls the development of malignant tumors (Review). Int J Oncol. 2023 Sep 1;63(3):1–10.
• Park SH, Baek KH, Shin I, Shin I. Subcellular Hsp70 Inhibitors Promote Cancer Cell Death via Different Mechanisms. Cell Chem Biol. 2018 Oct 18;25(10):1242-1254.e8.
• Cox A, Martini A, Ghozlan H, Moroose R, Zhu X, Lee E, et al. Chaperonin containing TCP1 as a marker for identification of circulating tumor cells in blood. PLoS One. 2022 Jun 24;17(6):e0264651.
• Doğan B, Pirim D, Işık Ö, Evrensel T. Candidate Biomarkers Associated With Circulating Tumor Cell Status in Metastatic Colorectal Cancer. J Clin Lab Anal. 2025 Apr;39(7):e70013.
• Lin D, Shen L, Luo M, Zhang K, Li J, Yang Q, et al. Circulating tumor cells: biology and clinical significance. Sig Transduct Target Ther. 2021 Nov 22;6(1):1–24.
• Vafaei S, Fattahi F, Ebrahimi M, Janani L, Shariftabrizi A, Madjd Z. Common molecular markers between circulating tumor cells and blood exosomes in colorectal cancer: a systematic and analytical review. Cancer Manag Res. 2019 Sep 25;11:8669–98.
• He K, Wang Z, Luo M, Li B, Ding N, Li L, et al. Metastasis organotropism in colorectal cancer: advancing toward innovative therapies. J Transl Med. 2023 Sep 9;21(1):612.
• Breuninger S, Stangl S, Werner C, Sievert W, Lobinger D, Foulds GA, et al. Membrane Hsp70-A Novel Target for the Isolation of Circulating Tumor Cells After Epithelial-to-Mesenchymal Transition. Front Oncol. 2018;8:497.
• Arias E, Cuervo AM. Pros and Cons of Chaperone-Mediated Autophagy in Cancer Biology. Trends Endocrinol Metab. 2020 Jan;31(1):53–66.
• Liu, H., Chen, L., Chen, Y. et al. TCP1 promotes the progression of malignant tumours by stabilizing c-Myc through the AKT/GSK-3β and ERK signalling pathways. Commun Biol 8, 563 (2025). https://doi.org/10.1038/s42003-025- 07867-6
• Liu Q, Qi Y, Kong X, Wang X, Zhang W, Zhai J, et al. Molecular and Clinical Characterization of CCT2 Expression and Prognosis via Large-Scale Transcriptome Profile of Breast Cancer. Front Oncol. 2021;11:614497.
• Lv W, Shi L, Pan J, Wang S. Comprehensive prognostic and immunological analysis of CCT2 in pan-cancer. Front Oncol. 2022;12:986990.
• Ahmed MZ, Billah MM, Ferdous J, Antar SI, Al Mamun A, Hossain MJ. Pan-cancer analysis reveals immunological and prognostic significance of CCT5 in human tumors. Sci Rep. 2025 Apr 24;15(1):14405.
• hang Z, Wang ZX, Chen YX, Wu HX, Yin L, Zhao Q, et al. Integrated analysis of single-cell and bulk RNA sequencing data reveals a pan-cancer stemness signature predicting immunotherapy response. Genome Med. 2022 Apr 29;14(1):45.
• Li J, Xu N, Hu L, Xu J, Huang Y, Wang D, et al. Chaperonin containing TCP1 subunit 5 as a novel pan-cancer prognostic biomarker for tumor stemness and immunotherapy response: insights from multi-omics data, integrated machine learning, and experimental validation. Cancer Immunol Immunother. 2025 May 27;74(7):224.
• Xanthopoulos A, Samt AK, Guder C, Taylor N, Roberts E, Herf H, et al. Hsp70—A Universal Biomarker for Predicting Therapeutic Failure in Human Female Cancers and a Target for CTC Isolation in Advanced Cancers. Biomedicine (Taipei). 2023 Aug;11(8):2276.
• Zhao K, Zhou G, Liu Y, Zhang J, Chen Y, Liu L, et al. HSP70 Family in Cancer: Signaling Mechanisms and Therapeutic Advances. Biomolecules. 2023 Apr;13(4):601.
• Abi Zamer B, El-Huneidi W, Eladl MA, Muhammad JS. Ins and Outs of Heat Shock Proteins in Colorectal Carcinoma: Its Role in Carcinogenesis and Therapeutic Perspectives. Cells. 2021 Oct 24;10(11):2862.
• Gao G, Liu S, Yao Z, Zhan Y, Chen W, Liu Y. The Prognostic Significance of Hsp70 in Patients with Colorectal Cancer Patients: A PRISMA-Compliant Meta-Analysis. Biomed Res Int. 2021 Apr 16;2021:5526327.
• Gráf L, Barabás L, Madaras B, Garam N, Maláti É, Horváth L, et al. High serum Hsp70 level predicts poor survival in colorectal cancer: Results obtained in an independent validation cohort. Cancer Biomark. 2018;23(4):539–47.