Treatment of ovarian cancer cell lines with 5-aza-2'-deoxycytidine upregulates the expression of cancer-testis antigens and class I major histocompatibility complex-encoded molecules
Treatment of ovarian cancer cell lines with 5-aza-2'-deoxycytidine upregulates the expression of cancer-testis antigens and class I major histocompatibility complex-encoded molecules
复制标题
DOI：
10.1007/s00262-008-0582-6
复制
发表时间：
2009-04-01
影响因子：
5.8
通讯作者：
Hogan, Kevin T.
中科院分区：
文献类型：
--
作者：
Adair, Sara J.;Hogan, Kevin T.
为了检验DNA甲基化降低将增加卵巢癌细胞的癌-睾丸抗原（CTA）和I类主要组织相容性复合体（MHC）编码分子的表达的假设，并因此增加这些细胞被抗原反应性CD 8（+）T细胞识别的能力。人卵巢癌细胞系在存在或不存在不同浓度的DNA去甲基化剂5-氮杂-2 ′-脱氧胞苷（DAC）处理3-7天。采用聚合酶链反应检测12个CTA基因的表达水平。用流式细胞术检测I类MHC分子和MAGE-A1蛋白表达水平。使用干扰素-γ ELISpot分析测定T细胞反应性dDAC处理卵巢癌细胞系增加了测试的12种CTA基因中的11种的表达，包括MAGE-A1、MAGE-A3、MAGE-A4、MAGE-A6、MAGE-A10、MAGE-A12、NY-ESO-1、TAG-1、TAG-2a、TAG-2b和TAG-2c。相比之下，DAC治疗降低了卵巢癌细胞已经很低的MAGE-A2基因表达，这是以前在任何组织学类型的癌症中都没有观察到的发现。DAC处理增加了细胞I类MHC分子的表达。这些作用在7天的间隔内是时间依赖性的，并且对于CTA高达1-3 μ M和对于I类MHC分子高达10 μ M是剂量依赖性的。每个测试的细胞系在暴露于DAC后具有独特的基因上调模式。增强的表达水平增加了抗原反应性CD 8（+）T细胞识别的3种抗原中的2种的识别。这些结果证明了DAC治疗与疫苗治疗相结合的潜在效用，试图诱导疫苗靶向的抗原的表达，但它们也证明了必须小心靶向诱导抗原。
To test the hypothesis that decrease in DNA methylation will increase the expression of cancer-testis antigens (CTA) and class I major histocompatibility complex (MHC)-encoded molecules by ovarian cancer cells, and thus increase the ability of these cells to be recognized by antigen-reactive CD8(+) T cells.Human ovarian cancer cell lines were cultured in the presence or absence of varying concentrations of the DNA demethylating agent 5-aza-2'-deoxycytidine (DAC) for 3-7 days. The expression levels of 12 CTA genes were measured using the polymerase chain reaction. The protein expression levels of class I MHC molecules and MAGE-A1 were measured by flow cytometry. T cell reactivity was determined using interferon-gamma ELISpot analysis.DAC treatment of ovarian cancer cell lines increased the expression of 11 of 12 CTA genes tested including MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE-A12, NY-ESO-1, TAG-1, TAG-2a, TAG-2b, and TAG-2c. In contrast, DAC treatment decreased the already low expression of the MAGE-A2 gene by ovarian cancer cells, a finding not previously observed in cancers of any histological type. DAC treatment increases the expression of class I MHC molecules by the cells. These effects were time-dependent over a 7-day interval, and were dose-dependent up to 1-3 mu M for CTA and up to 10 mu M for class I MHC molecules. Each cell line tested had a unique pattern of gene upregulation after exposure to DAC. The enhanced expression levels increased the recognition of 2 of 3 antigens recognized by antigen-reactive CD8(+) T cells.These results demonstrate the potential utility of combining DAC therapy with vaccine therapy in an attempt to induce the expression of antigens targeted by the vaccine, but they also demonstrate that care must be taken to target inducible antigens.