Increased genome instability and oxidative DNA damage and their association with IGF-1 levels in patients with active acromegaly

doi:10.1016/j.ghir.2013.12.002

Growth Hormone & IGF Research

Volume 24, Issue 1, February 2014, Pages 29-34

https://doi.org/10.1016/j.ghir.2013.12.002 Get rights and content

Abstract

Objective

The objectives of this study were to assess cytokinesis-block micronucleus cytome (CBMN Cyt) assay parameters and also oxidative DNA damage in patients with active acromegaly and controls and to assess the relationship between age, serum insulin-like growth factor 1 (IGF-1) levels, pituitary adenoma diameters, 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels and CBMN Cyt assay parameters in patients with active acromegaly.

Design

The study population included 30 patients with active acromegaly and 30 age- and sex-matched healthy controls. CBMN Cyt assay parameters in peripheral blood lymphocytes of patients with active acromegaly and controls were evaluated and plasma 8-OHdG levels were measured.

Results

Frequencies of micronucleus (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) in lymphocytes of patients with acromegaly were found to be significantly higher than those in controls (p < 0.001, p < 0.001, p < 0.001, respectively). The frequencies of apoptotic and necrotic cells in lymphocytes of patients with acromegaly were found to be significantly higher than those in controls (p < 0.001 and p < 0.001 respectively). No statistically significant differences in the number of cells in metaphase, the number of bi-nucleated cells (M2), the number of tri-nucleated cells (M3), the number of tetra-nucleated cells (M4) and nuclear division index (NDI) values were observed between patients and controls (p > 0.05). Plasma 8-OHdG (ng/ml) levels in patients with acromegaly were found to be significantly higher than those in controls (p < 0.005). MN frequency in the lymphocytes of patients with acromegaly increased with elevated serum IGF-1 levels (p < 0.05), whereas the number of NPBs and the frequency of apoptotic cells decreased with elevated serum IGF-1 levels (p < 0.01 and p < 0.05 respectively).

Conclusions

Both the increase in chromosomal/oxidative DNA damage and the positive association between MN frequency and serum IGF-1 levels may predict an increased risk of malignancy in acromegalic patients. Long-term follow-up of patients with acromegaly will be necessary to establish the degree of cancer risk in this population.

Introduction

Acromegaly is a slow-developing disease, with an estimated incidence of 4–5 per million per year and a prevalence of 60 cases per million [1], [2], [3]. It is usually caused by pituitary somatotroph adenomas and is associated with an increase in morbidity and mortality often attributed to cardiovascular, cerebrovascular, respiratory and metabolic diseases [4], [5], [6]. It is reported that acromegaly patients have an increased risk of developing both benign and malignant tumours, particularly colorectal cancer, and possibly other cancers such as breast, prostate, thyroid and haematological malignancies [7], [8]. The aetiology of these cancers remains uncertain, but they may be related to both growth hormone (GH) and insulin-like growth factor 1 (IGF-1) levels [9], [10], [11], [12], [13], [14].

However, some epidemiological studies report conflicting findings on cancer risk in acromegaly [15], [16]. It is established that IGF-1 is a potent mitogen and anti-apoptotic for a large variety of cells, exerting its mitogenic action by increasing DNA synthesis and stimulating cell cycle progression, whereas the main IGF-binding protein (IGFBP)-3 is a potent inhibitor of IGF-I action, which works in part by binding IGF-1 and also by inducing apoptosis through an IGF-1-independent mechanism in target cells [14], [15], [16], [17], [18]. It is well known that GH promotes the stimulation of both IGF-1 and IGFBP-3. Because IGF-1 stimulates cell proliferation/growth advantage and IGFBP-3 promotes apoptosis and because GH stimulates both IGF-1 and IGFBP-3 in acromegaly, these factors should balance cell growth regulation [16], [18]. However, high levels of IGFBP-3 were associated with an increased risk of premenopausal breast cancer, but not with any other cancer [17].

The micronucleus (MN) assay and cytokinesis-block micronucleus (CBMN) assay have both been used as methods for evaluating chromosome damage in cultured human lymphocytes because they provide a convenient and reliable index of both chromosome breakage and chromosome loss [19], [20]. An important and further development in the CBMN assay is the adoption of the CBMN cytome (CBMN Cyt) assay approach that scores MN and includes other nuclear abnormalities such as nuclear buds (NBUDs) and nucleoplasmic bridges (NPBs), as well as involve frequencies of necrotic and apoptotic cells and the proportion of dividing cells [21], [22]. This method allows the measurement of MN as a biomarker of chromosome breakage and/or whole chromosome loss, NPBs as a biomarker of dicentric chromosomes resulting from telomere end-fusions or DNA misrepair, NBUDs as a biomarker of gene amplification, necrosis and apoptosis as a viability status of cell, and metaphase, anaphase, mono-nucleated, bi-nucleated (BN), and multi-nucleated cells as a mitotic status of cell [21], [22].

Oxidative stress is known to potentially cause DNA damage involving point mutations because of base oxidation, single and double strand breaks and genomic instability. 8-Hydroxy-2′-deoxyguanosine (8-OHdG) is one of the damaged DNA products caused by reactive oxygen species. 8-OHdG is used as a reliable marker of oxidative DNA damage [23], [24], [25].

Our previous study showed that agromegalic patients had increased MN frequency [26]. However, the frequencies of NBPs and NBUDs, which determine DNA damage effects, the proportions of necrotic and apoptotic cells, which determine cytotoxicity and the proportions of mono-, bi- and multi-nucleated cells, and nuclear division index (NDI), which determines cytostasis in patients with acromegaly have not been investigated. The main objectives of this study are to assess the MN frequencies and new criteria such as the frequencies of NPBs and NBUDs, the frequencies of necrotic and apoptotic cells and NDI using CBMN Cyt assay parameters and oxidative DNA damage using the 8-OHdG assay in patients with acromegaly as well as to assess the relationship between age, serum IGF-1 levels, pituitary adenoma diameters, 8-OHdG levels and CBMN Cyt assay parameters, to improve the diagnosis and treatment of acromegaly.

Section snippets

Patients and controls

The study was performed on 30 patients diagnosed with active acromegaly and admitted to the Department of Endocrinology and Metabolism at Erciyes University Medical Faculty from December 2010 to December 2011. Thirty age- and sex-matched healthy controls were also included in the study.

The diagnosis of acromegaly was made based on clinical and biochemical findings. Clinical diagnosis was made according to typical disfigurement of the patient related to progressive acral enlargement and

Results

Table 2 shows the statistical results and mean ± SD values of age, IGF-1 levels, 8-OHdG levels and CBMN Cyt assay parameters in 30 patients with acromegaly and 30 healthy controls.

Pituitary adenoma diameters of patients with acromegaly ranged from 3 to 40 mm (17.74 ± 10.14). The nadir GH levels of patients with acromegaly varied from 1.03 to 24 μIU/ml (6.77 ± 6.03). The serum IGF-1 levels of patients with acromegaly were between 343 and 1600 mU/l (811.27 ± 323.55), while the serum IGF-1 levels of control

Discussion

Acromegaly is characterised by excessive levels of GH and IGF-1. High concentrations of circulating IGF-1 have been associated with an increased risk of breast, prostate, colorectal cancer and haematological malignancies in humans in several epidemiological studies [9], [10], [11], [32], [33], [34], [35]. IGF-1 mediates growth, apoptosis and metastasis responses in cancer [11].

CBMN assay in peripheral blood lymphocytes is one of the best validated cytogenetic tests for measuring DNA damage,

Conflict of interest

The authors declare that there is no conflict of interest.

Acknowledgements

This work was supported by Erciyes University Scientific Research Projects Units (Project number: TSD-10-3327).

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Increased genome instability and oxidative DNA damage and their association with IGF-1 levels in patients with active acromegaly

Abstract

Objective

Design

Results

Conclusions

Introduction

Section snippets

Patients and controls

Results

Discussion

Conflict of interest

Acknowledgements

Best Pract. Res. Clin. Endocinol. Metab.

Cytokine Growth Factor Rev.

Growth Horm. IGF Res.

Lancet

Mutat. Res.

Mutat. Res.

Metabolism

Endocr. Pract.

Mutat. Res.

Trends Endocrinol. Metab.

Trends Endocrinol. Metab.

Biochim. Biophys. Acta

Biochimie

Growth Horm. IGF Res.

Higher prevalence of clinically relevant pituitary adenomas confirmed

Clin. Endocrinol. (Oxf)

Epidemiology of acromegaly

Pituitary

Clinical manifestations and diagnosis of acromegaly

Int. J. Endocrinol.

Factors influencing mortality in acromegaly

J. Clin. Endocrinol. Metab.

Acromegaly

Orphanet J. Rare Dis.

Acromegaly associated with multiple tumors

Intern. Med.

The prevalence of benign and malignant tumors in patients with acromegaly at a single institute

Endocr. J.

The effects of insulin-like growth factors on tumorigenesis and neoplastic growth

Endocr. Rev.