An epigenetic mechanism for capecitabine resistance in mesothelioma.
An epigenetic mechanism for capecitabine resistance in mesothelioma.
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DOI：
10.1016/j.bbrc.2009.12.095
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发表时间：
2010-01-15
影响因子：
3.1
通讯作者：
Otterson, G. A.
中科院分区：
文献类型：
--
作者：
Kosuri, K. V.;Wu, X.;Wang, L.;Villalona-Calero, M. A.;Otterson, G. A.
间皮瘤是一种罕见的恶性肿瘤，其全球发病率持续上升。晚期疾病的治疗标准是静脉注射培美曲塞和顺铂。抗叶酸剂卡培他滨的疗效显著低于培美曲塞。胸苷酸合成酶（TS）、二氢嘧啶脱氢酶（DPD）和胸苷磷酸化酶（TP）之间的平衡对卡培他滨的疗效至关重要。将来自间皮瘤细胞系的DNA用亚硫酸氢盐处理并通过MS-PCR检测，获得RNA用于真实的时间PCR分析，并且获得蛋白质裂解物用于Western免疫印迹分析。MTT法检测5-aza-CdR预处理或未处理细胞与5′-脱氧-5-氟尿嘧啶（DFUR）、5-FU和培美曲塞的细胞毒性。最后，对4个间皮瘤样本和心包组织进行细胞外生长因子-1（ECGF-1）基因的亚硫酸氢盐测序。测试的四种细胞系之一（H290）被ECGF-1甲基化。这对应于通过真实的时间PCR和Western免疫印迹检测的TP表达的缺乏。1 μM 5-aza-CdR处理可增加H290细胞TP mRNA和蛋白的表达。TP的底物DFUR在甲基化细胞系中暴露于5-aza-CdR后递送时显示出增加的细胞毒性。当细胞暴露于5-FU或培美曲塞（含或不含5-aza-CdR）时，任何未甲基化细胞系均无差异。患者肿瘤样本显示，与正常心包相比，ECGF-1中甲基化CpG位点的数量增加。ECGF-1的甲基化导致TP的转录沉默，这可以解释卡培他滨缺乏任何作用，尤其是与培美曲塞相比时。
Mesothelioma is an uncommon malignancy whose global incidence continues to rise. The therapeutic standard for advanced disease is intravenous pemetrexed and cisplatin. The antifolate capecitabine is significantly less effective than pemetrexed. The balance between thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), and Thymidine Phosphorylase (TP) is critical to the efficacy of capecitabine. DNA from mesothelioma cell lines was bisulfite treated and examined by MS-PCR, RNA was obtained for real time PCR analysis, and protein lysates were obtained for Western immunoblot analysis. Cytotoxicity was assessed by MTT assay, comparing 5-aza-CdR pretreated or untreated cells with 5′-Deoxy-5-fluorouridine (DFUR), 5-FU, and pemetrexed. Finally bisulfite sequencing of the extracellular growth factor-1 (ECGF-1) gene was performed on 4 mesothelioma samples and pericardial tissue. One of the four cell lines tested (H290) was methylated for ECGF-1. This corresponded to a lack of TP expression by real time PCR and Western immunoblot. Treatment with 1 μM 5-aza-CdR increased TP mRNA and protein expression in H290. DFUR, the substrate for TP, showed increased cytotoxicity when delivered after 5-aza-CdR exposure in the methylated cell line. There was no difference in any of the unmethylated cell lines when cells were exposed to 5-FU or pemetrexed with or without 5-aza-CdR. Patient tumor samples revealed an increased number of methylated CpG sites in ECGF-1 compared to normal pericardium. Methylation of ECGF-1, leads to transcriptional silencing of TP and may explain the lack of any effect of capecitabine, especially when compared to pemetrexed.
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DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
6
作者：
Bueno, R;Appasani, K;Sugarbaker, D
通讯作者：
Sugarbaker, D
影响因子：
5
作者：
Price, B
通讯作者：
Price, B
影响因子：
158.5
作者：
Hegi, ME;Diserens, A;Stupp, R
通讯作者：
Stupp, R
影响因子：
45.3
作者：
van Meerbeeck, JP;Gaafar, R;Giaccone, G
通讯作者：
Giaccone, G