Identification and verification of heat shock protein 60 as a potential serum marker for colorectal cancer

Abstract

Colorectal cancer (CRC) is a major public health issue worldwide, and novel tumor markers may contribute to its efficient management by helping in early detection, prognosis or surveillance of disease. The aim of our study was to identify new serum biomarkers for CRC, and we followed a phased biomarker discovery and validation process to obtain an accurate preliminary assessment of potential clinical utility. We compared colonic tumors and matched normal tissue from 15 CRC patients, using two-dimensional difference gel electrophoresis (2D-DIGE), and identified 17 proteins that had significant differential expression. These results were further confirmed by western blotting for heat shock protein (HSP) 60, glutathione-S-transferase Pi, α-enolase, T-complex protein 1 subunit β, and leukocyte elastase inhibitor, and by immunohistochemistry for HSP60. Using mAbs raised against HSP60, we developed a reliable (precision of 5–15%) and sensitive (0.3 ng·mL−1) immunoassay for the detection of HSP60 in serum. Elevated levels of HSP60 were found in serum from CRC patients in two independent cohorts; the receiver-operating characteristic curve obtained in 112 patients with CRC and 90 healthy controls had an area under the curve (AUC) of 0.70, which was identical to the AUC of carcinoembryonic antigen. Combination of serum markers improved clinical performance: the AUC of a three-marker logistic regression model combining HSP60, carcinoembryonic antigen and carbohydrate antigen 19-9 reached 0.77. Serum HSP60 appeared to be more specific for late-stage CRC; therefore, future studies should evaluate its utility for determining prognosis or monitoring therapy rather than early detection.

The dissociation of the Hsp60/pro-Caspase-3 complex by bis(pyridyl)oxadiazole copper complex ( CubipyOXA ) leads to cell death in NCI-H292 cancer cells

“…Various data support the hypothesis that Hsp60 favors carcinogenesis and show a close correlation between high levels of Hsp60 and different types of cancer, thus the immunopositivity for Hsp60 can be considered a biomarker useful for the diagnosis and monitoring these types of malignancies [33,[49][50]. Frequently, the immunopositivity for Hsp60, in cancerous tissue or circulating cancer cells, correlates with a poor prognosis [49,[51][52][53].…”

Identification of aldolase A as a potential diagnostic biomarker for colorectal cancer based on proteomic analysis using formalin-fixed paraffin-embedded tissue

“…These criteria led us to select aldolase A, cyclophilin A, and annexin A2 as candidates. We did not include HSP60, neutrophil defensin1, or alpha-actinin-1, because previous reports had already suggested that these proteins might be biomarkers for CRC [26–29]. …”

Mitochondrial dysfunction in cancer: Potential roles of ATF5 and the mitochondrial UPR

“…For example, the mitochondrial chaperone HSP60 protects tumor cells from Bax-dependent cell death and CypD-dependent cell death by regulating mitochondrial permeability transition [116, 117]. HSP60 is also positively associated with tumor progression and hormone resistance in prostate cancer [118], and tumor differentiation in colorectal cancer [119]. mtHSP70, or mortalin, promotes tumor cell survival and epithelial-to-mesenchymal transition (EMT) by regulating the activities of p53 and PI3K–Akt pathways [120, 121], as well as facilitating metastasis of breast carcinoma and hepatocellular carcinoma [122, 123].…”