Remodeling of phospholipid composition in colon cancer cells by 1α,25(OH)2D3 and its analogs

doi:10.1016/j.jsbmb.2015.01.018

The Journal of Steroid Biochemistry and Molecular Biology

Volume 148, April 2015, Pages 172-178

https://doi.org/10.1016/j.jsbmb.2015.01.018 Get rights and content

Highlights

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1,25(OH)₂D₃ and its analogs modulated the lipid profile of human colon cancer cells.
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1,25(OH)₂D₃ and its analogs affected lipogenic gene expression in these cells.
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Analogs mostly induced similar and more pronounced changes compared to 1,25(OH)₂D₃.
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Changes in lipid profile after 1,25(OH)₂D₃/analog treatment were cell-line dependent.

Abstract

Alterations in cellular phospholipid composition are emerging as important traits in the development and progression of cancer. In this study we investigated the effects of 1α,25-dihydroxyvitamin D₃ [1,25(OH)₂D₃] and two of its more antiproliferative analogs on the cellular phospholipid composition of various human colon cancer cell lines. Treatment of Caco-2, SW1417 and SW480-ADH cells with 3 × 10⁻⁸ M 1,25(OH)₂D₃, CD578 or WU515 evoked significant changes in phospholipid composition, with the analogs being more potent than the natural compound. Observed effects included changes in acyl chain elongation and acyl chain saturation, and were substantially different in the various cell lines. Consistent with the alterations in phospholipid profiles, 1,25(OH)₂D₃ and its analogs provoked changes in several lipogenic enzymes such as fatty acid synthase (FASN), acetyl-CoA carboxylase (ACACA) and fatty acid elongases (ELOVLs). These effects were also cell line dependent. Taken together these findings indicate that 1,25(OH)₂D₃ and its analogs have divergent effects on the phospholipid composition of different colon cancer cell lines and warrant further investigation of the effect of 1,25(OH)₂D₃ and its analogs on lipid metabolism in various subtypes of primary human colon cancers.

Introduction

Cancer cells are characterized by rapid cell proliferation and changing microenvironments, and therefore undergo metabolic changes. Enhancing their endogenous lipid production is one of the metabolic adaptations made by cancer cells. Cancer cells use these newly synthesized fatty acids predominantly as building blocks for phospholipids for cellular membranes [1]. The lipid membrane mainly consists of phosphatidylcholine (PC) followed by phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS). These phospholipid classes contribute to proliferative growth and apoptosis of cells and act as precursors of many lipid second messenger molecules [2]. Moreover, as the newly synthesized fatty acids are mainly saturated and mono-unsaturated, increased lipid production also changes membrane phospholipid composition. In fact, saturated and mono-unsaturated phospholipids are packed more densely, which affects the structure and functioning of cellular membranes and attenuates the uptake of chemotherapeutics among others [3]. These lipids are also less vulnerable to lipid peroxidation and hence protect cells from oxidative stress and induction of apoptosis [3]. Many tumors overexpress lipogenic enzymes [4]. These include fatty acid synthase (FASN), which catalyzes the synthesis of saturated fatty acids. Also acetyl-CoA carboxylase alpha (ACACA), which produces malonyl-CoA for fatty acid synthesis and for further elongation of fatty acids by the fatty acid elongase (ELOVL) family of enzymes, is often overexpressed or activated. Likewise, mono-unsaturation by stearoyl-CoA desaturases (SCD) is active in many tumors [5], [6]. The overexpression of these enzymes has been linked to worsened prognosis and shorter disease-free survival in several tumor types [7]. For instance, overexpression of FASN is linked to short term survival in colorectal cancer [8] and correlates with higher risk of recurrence and a worse prognosis in breast cancer patients [9]. Inhibition of fatty acid synthesis by targeting FASN or ACACA is cytotoxic for cancer cells while normal cells remain unaffected [10].

The biologically active form of vitamin D₃, 1α,25-dihydroxyvitamin D₃ [1,25(OH)₂D₃], has several anti-tumor properties such as antiproliferative and prodifferentiating effects, as well as effects on invasion, metastasis and angiogenesis of tumors. Recently, it was shown that among the many actions of 1,25(OH)₂D₃, it inhibits FASN expression [11] by stimulation of long-chain fatty-acid-CoA ligase 3 [12] and alters fatty acid metabolism in LNCaP prostate cancer cells [13]. Also in ovarian cancer cell xenografts a non-calcemic vitamin D analog with anti-tumor properties was shown to decrease the expression of FASN as well as ACACA [14]. These studies prompted us to investigate the effects of 1,25(OH)₂D₃ and two of its analogs on the cellular phospholipid composition and the accompanying changes in lipogenic genes in three human colon cancer cell lines (Caco-2, SW1417 and SW480-ADH) that were selected for their different characteristics and dissimilar genetic abnormalities. SW1417 and SW480-ADH cells have respectively mutations in the BRAF and KRAS gene leading to abnormal epidermal growth factor receptor (EGFR) signaling. Caco-2 cells on the other hand, harbor mutations in genes important for Wnt/β-catenin and transforming growth factor-beta (TGF-β) signaling. Here, we report that 1,25(OH)₂D₃ and its analogs do affect the lipid profile of human colon cancer cells. The effects were cell line-dependent and were more outspoken for the analogs than for the mother compound.

Section snippets

Chemicals and reagents

1,25(OH)₂D₃ was obtained from Sigma–Aldrich (St. Louis, MO, USA). Vitamin D analogs 17-methyl-19-nor-21-nor-23-yne-26,27-F₆-1,25(OH)₂D₃ (CD578) and 23-yne-26,27-F₆-1,25(OH)₂D₃ (WU515) [15] were synthesized by M. Vandewalle and P. De Clercq (University of Ghent, Ghent, Belgium).

Cell culture

Caco-2 cells were obtained from the American Type Culture Collection (ATCC). SW1417, SW480-ADH and SW480-R cells were a kind gift of A. Muñoz (Universidad Autónoma de Madrid, Madrid, Spain). Caco-2, SW1417 and SW480-R

Vitamin D analogs induced stronger antiproliferative effects in colon cancer cells than 1,25(OH)₂D₃

In an earlier report, it was demonstrated that in MCF7 breast cancer cells two CD-ring modified vitamin D analogs, CD578 and WU515, have a 20-fold and 50-fold stronger antiproliferative effect than the mother compound [15]. In this study we investigated the antiproliferative effects of 1,25(OH)₂D₃ and these analogs in three different human colon cancer cell lines (Caco-2, SW1417 and SW480-ADH). In all tested cell lines cell proliferation was inhibited when cells were treated with a high

Discussion

1,25(OH)₂D₃ and its analogs have different antineoplastic effects on tumor cells in vitro and in vivo. These compounds can influence proliferation, differentiation, apoptosis, angiogenesis, metastasis and invasion of several types of cancer [16], [17], [18]. Cancer development and progression is often associated with increased de novo lipogenesis, which is mediated by oncogene and tumor suppressor gene-driven activation of lipogenic enzymes [19]. In this study we show that 1,25(OH)₂D₃ and its

Conflict of interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the article reported.

Acknowledgements

We thank Biauw Keng Tan, Suzanne Marcelis and Ine Beullens for excellent technical assistance. E.M. is recipient of a fellowship of the Agency for Innovation by Science and Technology IWT-Flanders. This work was supported by grants from the University of Leuven (GOA/14/010 (to A.V.) and GOA/11/2009 (to J.S.)) and Research Foundation-Flanders (FWO G.0587.09, G.0859.11 (to A.V.) and G0691.12 (to J.S.)).

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When triple negative breast cancer cells were treated with metformin (an anti-diabetic drug) growth arrest of the tumor cells was seen while also reducing the expression of ACACA (Wahdan-Alaswad et al., 2014). In another study on the colon cancer cell line, CaC0–2, it was observed that there is an increased expression of FASN and ACACA, but when the cell lines were treated with 1α,25-dihydroxy vitamin D3 [1,25(OH)2 D3] decline in proliferation and a decreased expression of ACACA were observed (Leyssens et al., 2015). The prostate cancer cell line PC-3 has an increased expression of ACACA but when the cell line was treated with quercetin, which is a dietary flavonoid,the expression of ACACA was found to be decreased and cell proliferation was inhibited, without cytotoxicity (Noori-Daloii et al., 2011).
Altered lipid metabolism is a characteristic feature of cancer cells. This systematic review assesses the risk for cancer development in association with the expression levels of the ACACA and ACACB genes which encode the rate limiting enzymes of fatty acid synthesis, acetyl-CoA carboxylase α (ACCα) and acetyl-CoA carboxylase β (ACCβ). The literature databases PubMed and Google Scholar were searched for variation in incidence of cancer depending on the expression of the ACACA and ACACB genes. In addition, related articles were retrieved from OMIM and Cochrane Library. Cohort studies, case control studies, studies using in vivo and in vitro models and randomized trials were evaluated to determine the risk for cancer associated with the altered expression of the genes. Overexpression of ACACA has been reported in cancers of the breast, lung, prostate, gliomas and colorectal cancer while low expression of ACACB has been reported in laryngeal squamous cell carcinoma, triple negative breast cancer, retinoblastoma, thyroid and melanoma. Increased expression of both ACACA and ACACB has been reported in hepatocellular carcinoma and glioblastoma. Various ACC-inhibitors with therapeutic potential have been developed, and several studies report the treatment of obesity related cancer by inhibiting ACC expression. Thus, ACC-inhibitors are potential therapeutic agents for effective treatment of cancer, especially obesity related cancers.
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al. (Jiang, Dai et al. 2015) reported the overexpression of ACACA in OVCA. In another study, it has been identified that non-calcemic vitamin D analog with anti-tumor properties has been shown to decrease ACACA expression in OVCA cell xenografts which further validated that ACACA overexpressed in OVCA (Leyssens, Marien et al. 2015). The ACLY gene is found as upregulated in several types of cancers, and is a key enzyme that drives the proliferation of cancer cells (Zaidi, Swinnen et al. 2012).
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ReviewRemodeling of phospholipid composition in colon cancer cells by 1α,25(OH)2D3 and its analogs

Highlights

Abstract

Introduction

Section snippets

Chemicals and reagents

Cell culture

Vitamin D analogs induced stronger antiproliferative effects in colon cancer cells than 1,25(OH)2D3

Discussion

Conflict of interest

Acknowledgements

Hum. Pathol.

J. Steroid Biochem. Mol. Biol.

FEBS Lett.

J. Steroid Biochem. Mol. Biol.

Chem. Biol.

Prog. Lipid Res.

Dysregulated lipid metabolism in cancer

World J. Biol. Chem.

The role of phosphatidylcholine and choline metabolites to cell proliferation and survival

Crit. Rev. Biochem. Mol. Biol.

De novo lipogenesis protects cancer cells from free radicals and chemotherapeutics by promoting membrane lipid saturation

Cancer Res.

Enzymes of the fatty acid synthesis pathway are highly expressed in in situ breast carcinoma

Clin. Cancer Res.