Colston KW, Perks CM, Xie SP and Holly JMGrowth inhibition of both MCF-7 and Hs578T human breast cancer cell lines by vitamin D analogues is associated with increased expression of insulin-like growth factor binding protein-3. J Mol Endocrinol 20: 157-162

Division of Gastroenterology, Endocrinology and Metabolism, St George's Hospital Medical School, London, UK.
Journal of Molecular Endocrinology (Impact Factor: 3.08). 02/1998; 20(1):157-62.
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The effects of two vitamin D analogues, EB1089 and CB1093, on insulin-like growth factor binding protein (IGFBP) expression have been examined in MCF-7 and Hs578T human breast cancer cell lines. Both vitamin D analogues inhibited IGF-1 stimulated growth of MCF-7 cells and enhanced the production of IGFBP-3 as determined by Western-ligand blotting. Recombinant human IGFBP-3 inhibited the growth of MCF-7 cells over the concentration range 1-235 ng/ml. Hs578T cells were unresponsive to the mitogenic effects of IGF-1 but growth was inhibited by the two vitamin D analogues. Treatment of Hs578T cells with EB1089 and CB1093 (10 nM) as well as 100 nM 9-cis retinoic acid (9-cis RA) or all-trans retinoic acid (ATRA) was associated with increased accumulation of IGFBP-3 in conditioned medium. Furthermore, cotreatment of Hs578T cells with EB1089 and 9-cis RA led to augmented effects on both inhibition of cell growth and IGFBP-3 accumulation in conditioned medium as assessed by Western ligand blotting and radioimmunoassay. These findings suggest a role for IGFBP-3 in the growth inhibitory effects of vitamin D analogues.

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    • "Positive regulators Tumour suppressor p53 Colon cancer cells [10] Cytokines, ligands and receptors Vitamin D Cervical and prostate cancer cells [5] [9] [22] [112] Retinoids Ectocervical and breast epithelial cells [51] [54] TGF-β1 H u m a n fibroblasts [93] [94] Breast and prostate cancer cells [51] [107] [119] TNF-α MDA-MB-231 cells [123] Chemo-therapeutics Anti-oestrogens MCF-7 cells [60] Paclitaxel Breast and gastric cancer cells [41] [76] HDAC inhibitors Liver cancer cells [84] Demethylating agents Breast cancer cells [147] Topoisomerase II inhibitors Breast, lung, gastric and prostate cancer cells [41] [49] [76] Environmental Hypoxia Embryonic stem cells [36] Squamous cancer cells (oesophageal) [104] Pancreatic cancer cells [67] Keratinocytes and fibroblasts (normal and transformed) [30] Brain, colon, lung, breast, cervical osteosarcoma and prostate cancer cells [49] [72] IR Endothelial cells [134] Squamous cell carcinoma cells [68] Negative regulators Tumour suppressor PTEN Gastric cancer cells [144] DNA methylation IGFBP-3 promoter Hepatocellular carcinoma [52] [146] Cytokines, ligands and receptors INF-γ Prostate cancer cells [35] Positive ER expression Breast epithelial cancer cells [17,59,60,131,143] Fig. 1. Comparison of the in vitro pro-survival and growth-promoting actions of IGFBP-3 with its pro-apoptotic mechanisms. "
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    ABSTRACT: One of the hallmarks in the advancement of cancer cells is an ability to overcome and acquire resistance to adverse conditions. There has been a large amount of cancer research on IGFBP-3 as a pro-apoptotic molecule in vitro. These pro-apoptotic properties, however, do not correlate with several studies linking high IGFBP-3 levels in breast cancer tissue to rapid growth and poor prognosis. Evidence is emerging that IGFBP-3 also exhibits pro-survival and growth-promoting properties in vitro. How IGFBP-3 pivots cell fate to either death or survival, it seems, comes down to a complex interplay between cells’ microenvironments and the presence of cellular IGFBP-3 binding partners and growth factor receptors. The cytoprotective actions of IGFBP-3 are not restricted to cancer but are also observed in other disease states, such as retinopathy and brain ischaemia. Here we review the literature on this paradoxical nature of IGFBP-3, its pro-apoptotic and growth-inhibitory actions versus its cytoprotective and growth-potentiating properties, and discuss the implications of targeting IGFBP-3 for treatment of disease.
    Growth Hormone & IGF Research 10/2014; 24(5). DOI:10.1016/j.ghir.2014.04.007 · 1.41 Impact Factor
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    • "0.5 × 103 cells were seeded in 96-well plate with serum-containing medium for 24 hours starvation. Different concentrations of either commercial rhIGFBP-3 (Sigma) or freshly extracted rhIGFBP-3 protein were added to MCF-7 cells on every 72 hours for a total of 9 days and the cells were finally collected for MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay [35]. HT-29 cells were grown in EMEM (Invitrogen) with 2 mM Glutamine, 1% Non Essential Amino acids (NEAA) and 10% heat-inactivated FBS (Hyclone) at 37°C in a humidified atmosphere with 5% CO2. "
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    ABSTRACT: Insulin-like growth factor binding protein-3 (IGFBP-3) is a multifunctional molecule which is closely related to cell growth, apoptosis, angiogenesis, metabolism and senescence. It combines with insulin-like growth factor-I (IGF-I) to form a complex (IGF-I/IGFBP-3) that can treat growth hormone insensitivity syndrome (GHIS) and reduce insulin requirement in patients with diabetes. IGFBP-3 alone has been shown to have anti-proliferation effect on numerous cancer cells. We reported here an expression method to produce functional recombinant human IGFBP-3 (rhIGFBP-3) in transgenic rice grains. Protein sorting sequences, signal peptide and endoplasmic reticulum retention tetrapeptide (KDEL) were included in constructs for enhancing rhIGFBP-3 expression. Western blot analysis showed that only the constructs with signal peptide were successfully expressed in transgenic rice grains. Both rhIGFBP-3 proteins, with or without KDEL sorting sequence inhibited the growth of MCF-7 human breast cancer cells (65.76 ± 1.72% vs 45.00 ± 0.86%, p < 0.05; 50.84 ± 1.97% vs 45.00 ± 0.86%, p < 0.01 respectively) and HT-29 colon cancer cells (65.14 ±3.84% vs 18.01 ± 13.81%, p < 0.05 and 54.7 ± 9.44% vs 18.01 ± 13.81%, p < 0.05 respectively) when compared with wild type rice. These findings demonstrated the feasibility of producing biological active rhIGFBP-3 in rice using a transgenic approach, which will definitely encourage more research on the therapeutic use of hIGFBP-3 in future.
    PLoS ONE 10/2013; 8(10):e77516. DOI:10.1371/journal.pone.0077516 · 3.23 Impact Factor
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    • "The ER-α negative cell lines secrete predominantly IGFBP-3 [18]. Furthermore IGFBP-3 is known to mediate the effects of a number of different inhibitors of cell growth and inducers of apoptosis, such as TGF-beta, p53, vitamin D and tamoxifen [19] [20] [21] [22] [23]. In this study we treated four different breast cancer cell lines with the DNA methytransferase inhibitor, AZA to determine the role played by IGFBP-3 in the effects of AZA on total cell number, cell survival and colony formation. "
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    ABSTRACT: Breast cancer progression is associated with loss of estrogen receptor(ER-α), often due to epigenetic silencing. IGFBP genes have consistently been identified among the most common to be aberrantly methylated in tumours. To understand the impact of losing IGFBP-3 tumour expression via DNA methylation, we treated four breast cancer cell lines (MCF-7, T47D, Hs578T and MDA-MB-231) with a DNA methyltransferase inhibitor, 5-Aza-2'-deoxycytidine(AZA) to determine IGFBP-3's role in the effects of AZA on total cell number and survival relative to changes in the ER. AZA induced cell growth inhibition, death and a reduction in the formation of colonies, despite increasing ER-α expression in ER-negative cells but reducing ER-α in ER-positive cells. Regardless of the differential effects on the ER-α, AZA consistently increased the abundance of IGFBP-3 and negating this increase in IGFBP-3 with siRNA reduced the AZA-induced growth inhibition and induction of cell death and virtually negated the AZA-induced inhibition of colony formation. With ER-α positive cells AZA increased the abundance of the tumour suppressor gene, p53 and induced demethylation of the IGFBP-3 promoter, whereas with ER negative cells, AZA epigenetically increased the transcription factor AP2-α, which when silenced prevented the increase in IGFBP-3. IGFBP-3 plays an important role in the anti-tumorigenic effects of AZA on breast cancer cells.
    Experimental Cell Research 06/2013; 319(14). DOI:10.1016/j.yexcr.2013.06.011 · 3.25 Impact Factor
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