Mild folate deficiency induces genetic and epigenetic instability and phenotype changes in prostate cancer cells.
Mild folate deficiency induces genetic and epigenetic instability and phenotype changes in prostate cancer cells.
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DOI：
10.1186/1741-7007-8-6
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发表时间：
2010-01-21
期刊：
影响因子：
5.4
通讯作者：
Smiraglia DJ
中科院分区：
文献类型：
--
作者：
Bistulfi G;Vandette E;Matsui S;Smiraglia DJ
叶酸（维生素B9）是细胞增殖所必需的，因为它参与脱氧胸苷一磷酸（dTMP）和s-腺苷甲硫氨酸（s-adenosylmethionine，sMet）的生物合成。叶酸消耗与上皮来源的癌症的发生和进展之间的联系具有高度的临床相关性，但仍不清楚。我们最近证实，对低叶酸利用率的敏感性受到多胺生物合成速率的影响，这在前列腺细胞中是突出的。因此，我们假设前列腺细胞可能对叶酸限制引起的遗传、表观遗传和表型变化高度敏感。我们研究了长期，轻度叶酸耗竭的后果，在一个模型，包括三个同源细胞系来源于转基因小鼠前列腺腺瘤（TRAMP）模型，概括不同阶段的前列腺癌;良性，转化和转移。高效液相色谱分析表明，轻度叶酸耗竭（100 nM）足以诱导所有前列腺细胞系中的核苷酸和蛋氨酸池的不平衡。随机寡核苷酸引物合成（ROPS）显示尿嘧啶错误掺入和DNA单链断裂的显着增加，而光谱核型分析（SKY）确定了五个新的染色体重排生长的细胞中轻度叶酸耗竭。使用全局方法，我们确定了叶酸消耗后CpG岛和组蛋白甲基化的增加，尽管总5-甲基胞嘧啶水平不变，表明叶酸消耗对表观遗传调控的广泛影响。这些基因组变化与前列腺细胞的表型变化相一致，包括增加的锚定非依赖性生长和降低的叶酸消耗敏感性。本研究表明，与组织培养中常规使用的超生理量叶酸（2 μ M）生长的细胞相比，前列腺细胞对轻度叶酸耗竭后的遗传和表观遗传变化高度敏感。此外，我们首次阐明了这些方面的贡献，从而在上皮细胞的表型变化。这些结果提供了一个强有力的理由，研究叶酸的操作对前列腺在体内的影响，其中细胞可能是更敏感的叶酸补充或抗叶酸治疗方法引起的叶酸状态的变化。
Folate (vitamin B9) is essential for cellular proliferation as it is involved in the biosynthesis of deoxythymidine monophosphate (dTMP) and s-adenosylmethionine (AdoMet). The link between folate depletion and the genesis and progression of cancers of epithelial origin is of high clinical relevance, but still unclear. We recently demonstrated that sensitivity to low folate availability is affected by the rate of polyamine biosynthesis, which is prominent in prostate cells. We, therefore, hypothesized that prostate cells might be highly susceptible to genetic, epigenetic and phenotypic changes consequent to folate restriction. We studied the consequences of long-term, mild folate depletion in a model comprised of three syngenic cell lines derived from the transgenic adenoma of the mouse prostate (TRAMP) model, recapitulating different stages of prostate cancer; benign, transformed and metastatic. High-performance liquid chromatography analysis demonstrated that mild folate depletion (100 nM) sufficed to induce imbalance in both the nucleotide and AdoMet pools in all prostate cell lines. Random oligonucleotide-primed synthesis (ROPS) revealed a significant increase in uracil misincorporation and DNA single strand breaks, while spectral karyotype analysis (SKY) identified five novel chromosomal rearrangements in cells grown with mild folate depletion. Using global approaches, we identified an increase in CpG island and histone methylation upon folate depletion despite unchanged levels of total 5-methylcytosine, indicating a broad effect of folate depletion on epigenetic regulation. These genomic changes coincided with phenotype changes in the prostate cells including increased anchorage-independent growth and reduced sensitivity to folate depletion. This study demonstrates that prostate cells are highly susceptible to genetic and epigenetic changes consequent to mild folate depletion as compared to cells grown with supraphysiological amounts of folate (2 μM) routinely used in tissue culture. In addition, we elucidate for the first time the contribution of these aspects to consequent phenotype changes in epithelial cells. These results provide a strong rationale for studying the effects of folate manipulation on the prostate in vivo, where cells might be more sensitive to changes in folate status resulting from folate supplementation or antifolate therapeutic approaches.
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