Mitochondrial creatine kinase in cancer patients

doi:10.3109/00313028709077128

Pathology

Volume 19, Issue 2, 1987, Pages 162-165

https://doi.org/10.3109/00313028709077128 Get rights and content

Summary

We have developed a quantitative immunoassisted enzyme assay to screen for creatine kinase isoenzymes BB (CK-BB) and mitochondrial (CK-m) using commercial antibodies and reagents. Presence of CK-m activity was subsequently confirmed by electrophoretic separation of samples with elevated values. A prospective clinical trial was undertaken in 117 subjects: Normal (30), cirrhosis patients (30), myocardial infarction patients (30), and untreated oncology patients with metastatic malignancy (27). In 12 patients with malignancy CK-m activities were elevated; all had adenocarcinomas. No significant activity was detected in patients with other malignancies. CK-m positive tumour patients had a significantly higher mortality rate, and in two instances death was preceded by a sudden rise in CK-m activity. We suggest CK-m is a marker of adenocarcinoma and its presence in serum signifies increased mortality.

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Cited by (30)

Overexpression of ubiquitous mitochondrial creatine kinase (uMtCK) accelerates tumor growth by inhibiting apoptosis of breast cancer cells and is associated with a poor prognosis in breast cancer patients
2012, Biochemical and Biophysical Research Communications
Citation Excerpt :
Overexpression of CK has been demonstrated in a variety of solid tumors and tumor cell lines [6–10]. Elevated CK has been detected in the serum of patients with advanced cancer and is associated with a poor prognosis [7,9,11]. Overexpression of uMtCK has been reported in several cancers with poor prognoses, such as lung cancer, gastric cancer and prostate cancer [6,7,12].
Ubiquitous mitochondrial creatine kinase (uMtCK), a mitochondrial isoenzyme of creatine kinase (CK), is a central controller of cellular energy homeostasis. Overexpression of uMtCK has been reported to be associated with a poor prognosis for several tumors. The aim of this study was to assess its association with breast cancer (BCa) and to further investigate its underlying mechanisms.
We first detected uMtCK expression by immunohistochemistry in human BCa tissues and assessed the association with the prognosis of patients. We then evaluated uMtCK expression in crowded and normal condition cultures of several human BCa cell lines. After two stable clones of the MDA-MB-231 cell line with high expression of uMtCK were established, cell growth, apoptosis and mitochondrial apoptotic pathway protein expression were measured in these clones. Finally, tumorigenicity of the above cells was assessed using nude mice to explore the relationship between uMtCK expression and tumor progression.
uMtCK expression was detected in 85.5% (47 of 55) of the invasive ductal carcinomas of breast tissue, not otherwise specified (IDC-NOS). Expression in BCa tissue was significantly associated with reduced progression-free survival (PFS; P = 0.019) and overall survival (OS; P = 0.022) of the patients. Up-regulation of uMtCK expression was identified in crowded BCa cells in culture, and the number of apoptotic cells was significantly decreased in uMtCK transfected MDA-MB-231 cell clones (P < 0.01). Stabilization of the mitochondrial membrane potential (ΔΨm) and down regulation of cytochrome c (cyt c) and activated caspase 9, two components of mitochondrial apoptotic pathway proteins, were also identified in the same clones when cells were crowded in culture. In vivo studies revealed that the transfected tumor cells with uMtCK overexpression induced faster tumor growth in nude mice, along with accelerated animal body weight loss and a significantly lower tumor apoptotic index (AI) (P < 0.001).
The results indicated that uMtCK expression is associated with a poor prognosis in BCa and might serve as a tumor marker. In vivo and In vitro evidence suggests that uMtCK overexpression promotes tumor growth by inhibiting apoptosis of tumor cells through stabilizing ΔΨm and down regulating mitochondrial apoptotic pathway proteins. Exploration of therapeutic agents targeting the expression of uMtCK may have practical value for BCa patients.
Mitochondrial creatine kinase in human health and disease
2006, Biochimica et Biophysica Acta - Molecular Basis of Disease
Citation Excerpt :
Some clinical studies [197,240], but not all [241], have shown that creatine supplementation may enhance function in patients with mitochondrial cytopathies. Overexpression of uMtCK has been reported for several tumors with poor prognosis [242–244]. Namely in a Hodgkin's disease cell line, uMtCK, but not cytosolic BB-CK, is one of the very few genes that is specifically up-regulated [245].
Mitochondrial creatine kinase (MtCK), together with cytosolic creatine kinase isoenzymes and the highly diffusible CK reaction product, phosphocreatine, provide a temporal and spatial energy buffer to maintain cellular energy homeostasis. Mitochondrial proteolipid complexes containing MtCK form microcompartments that are involved in channeling energy in form of phosphocreatine rather than ATP into the cytosol. Under situations of compromised cellular energy state, which are often linked to ischemia, oxidative stress and calcium overload, two characteristics of mitochondrial creatine kinase are particularly relevant: its exquisite susceptibility to oxidative modifications and the compensatory up-regulation of its gene expression, in some cases leading to accumulation of crystalline MtCK inclusion bodies in mitochondria that are the clinical hallmarks for mitochondrial cytopathies. Both of these events may either impair or reinforce, respectively, the functions of mitochondrial MtCK complexes in cellular energy supply and protection of mitochondria form the so-called permeability transition leading to apoptosis or necrosis.
C-terminal lysines determine phospholipid interaction of sarcomeric mitochondrial creatine kinase
2004, Journal of Biological Chemistry
Citation Excerpt :
This intimate exchange of substrates and products, the so-called functional coupling or metabolite channeling (10, 11), fulfills important functions that may vary among different tissues, species, and developmental states (12, 13): (i) phosphocreatine becomes the high energy intermediate that is exported from mitochondria into the cytosol (3); (ii) locally generated ADP stimulates oxidative phosphorylation (11); and (iii) ADP channeled through the MtCK/ANT interaction inhibits the Ca2+-induced opening of the mitochondrial permeability transition pore (14, 15), a well known trigger of apoptosis (16, 17). Thus, overexpression of uMtCK in many malignant cancers with especially poor prognosis (18, 19) may be related to high energy turnover and failure to eliminate cancer cells via apoptosis. MtCK functions in energy buffering and transport, as well as permeability transition pore regulation may also explain the supportive and protective effects of the CK substrate creatine in many muscular, neurodegenerative, and age-related disorders (20–22).
High affinity interaction between octameric mitochondrial creatine kinase (MtCK) and the phospholipid cardiolipin in the inner mitochondrial membrane plays an important role in metabolite channeling between MtCK and inner membrane adenylate translocator, which itself is tightly bound to cardiolipin. Three C-terminal basic residues revealed as putative cardiolipin anchors in the x-ray structures of MtCK and corresponding to lysines in human sarcomeric MtCK (sMtCK) were exchanged by in vitro mutagenesis (K369A/E, K379Q/A/E, K380Q/A/E) to yield double and triple mutants. sMtCK proteins were bacterially expressed, purified to homogeneity, and verified for structural integrity by enzymatic activity, gel filtration chromatography, and CD spectroscopy. Interaction with cardiolipin and other acidic phospholipids was quantitatively analyzed by light scattering, surface plasmon resonance, and fluorescence spectroscopy. All mutant sMtCKs showed a strong decrease in vesicle cross-linking, membrane affinity, binding capacity, membrane ordering capability, and binding-induced changes in protein structure as compared with wild type. These effects did not depend on the nature of the replacing amino acid but on the number of exchanged lysines. They were moderate for Lys-379/Lys-380 double mutants but pronounced for triple mutants, with a 30-fold lower membrane affinity and an entire lack of alterations in protein structure compared with wild-type sMtCK. However, even triple mutants partially maintained an increased order of cardiolipin-containing membranes. Thus, the three C-terminal lysines determine high affinity sMtCK/cardiolipin interaction and its effects on MtCK structure, whereas low level binding and some effect on membrane fluidity depend on other structural components. These results are discussed in regard to MtCK microcompartments and evolution.
Octamers of mitochondrial creatine kinase isoenzymes differ in stability and membrane binding
2000, Journal of Biological Chemistry
Citation Excerpt :
Preliminary results indicate that human uMtCK is in fact more resistant to peroxynitrite-induced octamer dissociation than sMtCK.3 The specific properties of uMtCK may also explain, for instance, why tumors overexpressing this isoenzyme are renowned for their especially poor prognosis (11, 12). Further experiments will be necessary to confirm this hypothesis.
Octamer stability and membrane binding of mitochondrial creatine kinase (MtCK) are important for proper functioning of the enzyme and were suggested as targets for regulatory mechanisms. A quantitative analysis of these properties, using fluorescence spectroscopy, gel filtration, and surface plasmon resonance, revealed substantial differences between the two types of MtCK isoenzymes, sarcomeric (sMtCK) and ubiquitous (uMtCK). As compared with human and chicken sMtCK, human uMtCK showed a 23–34 times slower octamer dissociation rate, a reduced reoctamerization rate and a superior octamer stability as deduced from the octamer/dimer ratios at thermodynamic equilibrium. Octamer stability of sMtCK increased with temperature up to 30 °C, indicating a substantial contribution of hydrophobic interactions, while it decreased in the case of uMtCK, indicating the presence of additional polar dimer/dimer interactions. These conclusions are consistent with the recently solved x-ray structure of the human uMtCK (Eder, M., Fritz-Wolf, K., Kabsch, W., Wallimann, T., and Schlattner, U. (2000) Proteins 39, 216–225). When binding to 16% cardiolipin membranes, sMtCK showed slightly faster on-rates and higher affinities than uMtCK. However, human uMtCK was able to recruit the highest number of binding sites on the vesicle surface. The observed divergence of ubiquitous and sarcomeric MtCK is discussed with respect to their molecular structures and the possible physiological implications.
Cloning, Escherichia coli expression, and phase-transition chromatography-based purification of recombinant rabbit heart mitochondrial creatine kinase
1999, Protein Expression and Purification
A cDNA clone of the mitochondrial sarcomeric creatine kinase cDNA was obtained by screening a rabbit heart library. This cDNA is characterized by a 1257-nucleotide open reading frame encoding a 419-amino-acid protein with a cleavable 39-amino-acid mitochondrial presequence (Accession No. AJ011334). This new member of the guanidino kinase family shows a high degree of sequence similarity with the other phosphagen kinases sequenced so far. The mature enzyme was efficiently expressed in Escherichia coli BL21(DE3) cells as a soluble octameric protein using the pET21 plasmid and purified by a three-step improved method including a final phase-transition chromatography.
Mitochondrial creatine kinase: a key enzyme of aerobic energy metabolism
1992, BBA - Bioenergetics

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Mitochondrial creatine kinase in cancer patients

Summary

Clin Chim Acta

High activity of CK in mitochondria from muscle and brain, and evidence of a separate isoenzyme of CK

Biochem Biophys Res Commun

Characteristics of mitochondrial creatine kinases from normal heart and liver

Clin Chim Acta

Human CK: Isoenzymes and logistics of energy distribution

Scand J Clin Lab Invest

The cell nucleus as a possible source of catho-dally migrating serum CK

Clin Chem

Mitochondrial CK as a tumour associated marker

Clin Chim Acta

Macro CK: A possible marker of GI cancer

Surgery

Serum CK isoenzymes in a patient with advanced malignant disease

Acta Med Scand

Cathodal CK band, a poor prognostic sign in malignancy

Clin Chem

Atypical cathodally migrating CK: A possible tumour associated enzyme marker for disseminated malignancy (Letter)

Clin Chem