Expression level and DNA methylation status of glutathione-S-transferase genes in normal murine prostate and TRAMP tumors.
Expression level and DNA methylation status of glutathione-S-transferase genes in normal murine prostate and TRAMP tumors.
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DOI：
10.1002/pros.20976
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发表时间：
2009-09-01
期刊：
影响因子：
2.8
通讯作者：
Karpf, Adam R.
中科院分区：
文献类型：
--
作者：
Mavis, Cory K.;Kinney, Shannon R. Morey;Foster, Barbara A.;Karpf, Adam R.
谷胱甘肽 - S - 转移酶（Gst）基因在人类前列腺癌中表达下调，并且在这种恶性肿瘤中，GSTP1基因沉默是由启动子DNA高度甲基化介导的。我们检测了正常小鼠前列腺和小鼠前列腺转基因腺癌（TRAMP）肿瘤中的Gst基因表达以及Gst启动子DNA甲基化情况。
从TRAMP小鼠获取原发性和转移性肿瘤，从品系匹配的野生型小鼠获取正常前列腺（每组n = 15）。采用实时定量逆转录 - 聚合酶链反应（qRT - PCR）测量GstA4、GstK1、GstM1、GstO1和GstP1的mRNA表达，采用蛋白质印迹法和免疫组织化学染色法测量GstM1和GstP1的蛋白质表达。采用MassARRAY定量甲基化分析测量GstA4、GstK1、GstM1、GstO1和GstP1的5’CpG岛的DNA甲基化情况。用表观遗传重塑药物地西他滨和曲古抑菌素A（TSA）单独及联合处理TRAMP - C2细胞，并测量Gst基因表达。
在所分析的基因中，GstM1和GstP1在正常前列腺中表达水平最高。与正常前列腺相比，所有5种Gst基因在原发性肿瘤中的表达均大幅降低，但在肿瘤转移灶中并非如此。与正常前列腺相比，TRAMP肿瘤中的Gst启动子甲基化没有变化。地西他滨 + TSA联合处理显著增强了TRAMP - C2细胞中4/5的Gst基因的表达。
Gst基因在原发性TRAMP肿瘤中广泛下调，但在转移性TRAMP肿瘤中没有。启动子DNA高度甲基化似乎并不驱动TRAMP原发性肿瘤中Gst基因的抑制；然而，利用TRAMP细胞进行的药理学研究表明，表观遗传机制参与了Gst基因的抑制。
Glutathione-S-transferase (Gst) genes are down-regulated in human prostate cancer, and GSTP1 silencing is mediated by promoter DNA hypermethylation in this malignancy. We examined Gst gene expression and Gst promoter DNA methylation in normal murine prostates and Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) tumors. Primary and metastatic tumors were obtained from TRAMP mice, and normal prostates were obtained from strain-matched WT mice (n=15/group). Quantitative real-time RT-PCR was used to measure GstA4, GstK1, GstM1, GstO1, and GstP1 mRNA expression, and Western blotting and immunohistochemical staining was used to measure GstM1 and GstP1 protein expression. MassARRAY Quantitative Methylation Analysis was used to measure DNA methylation of the 5’ CpG islands of GstA4, GstK1, GstM1, GstO1, and GstP1. TRAMP-C2 cells were treated with the epigenetic remodeling drugs decitabine and trichostatin A (TSA) alone and in combination, and Gst gene expression was measured. Of the genes analyzed, GstM1 and GstP1 were expressed at highest levels in normal prostate. All five Gst genes showed greatly reduced expression in primary tumors compared to normal prostate, but not in tumor metastases. Gst promoter methylation was unchanged in TRAMP tumors compared to normal prostate. Combined decitabine + TSA treatment significantly enhanced the expression of 4/5 Gst genes in TRAMP-C2 cells. Gst genes are extensively downregulated in primary but not metastatic TRAMP tumors. Promoter DNA hypermethylation does not appear to drive Gst gene repression in TRAMP primary tumors; however, pharmacological studies using TRAMP cells suggest the involvement of epigenetic mechanisms in Gst gene repression.
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影响因子：
11.2
作者：
Deeb, Kristin K.;Michalowska, Aleksandra M.;Green, Jeffrey E.
通讯作者：
Green, Jeffrey E.
影响因子：
3.3
作者：
Bostwick, David G.;Meiers, Isabelle;Shanks, Jonathan H.
通讯作者：
Shanks, Jonathan H.
影响因子：
3.9
作者：
Hayes, JD;Chanas, SA;Yamamoto, M
通讯作者：
Yamamoto, M
影响因子：
30.8
作者：
Cameron, EE;Bachman, KE;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
4.2
作者：
Fang, Mingzhu;Chen, Dapeng;Yang, Chung S.
通讯作者：
Yang, Chung S.