Survey of differentially methylated promoters in prostate cancer cell lines
Survey of differentially methylated promoters in prostate cancer cell lines
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DOI：
10.1593/neo.05289
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发表时间：
2005-08-01
期刊：
影响因子：
4.8
通讯作者：
McClelland, M
中科院分区：
文献类型：
--
作者：
Wang, YP;Yu, QJ;McClelland, M
使用基于微阵列的技术比较了三种永生化前列腺上皮细胞和五种癌细胞系（LNCaP、PC 3、PC 3 M、PC 3 M-Pro 4和PC 3 M-LN 4）的基因组中的DNA甲基化和拷贝数。用甲基化敏感酶HpaII切割基因组DNA，然后进行接头连接、聚合酶链式反应（PCR）扩增、标记和与启动子序列阵列杂交。只有那些在几百个碱基对内具有未甲基化限制性位点的基因组DNA部分才能产生在阵列上可检测的PCR产物。在测试阵列上的2732个启动子序列中，504个（18.5%）显示永生化前列腺上皮细胞系和癌细胞系之间的差异杂交。在癌症衍生系的候选高甲基化基因中，有8个（CD 44、CDKN 1A、ESR 1、PLAU、RARB、SFN、TNFRSF 6和TSPY）先前在前列腺癌中观察到，13个先前已知的甲基化靶点在其他癌症中（ARHI、bc 1 -2、BRCA 1、CDKN 2C、GADD 45 A、MTAP、PGR、SLC 26 A4、ALF 3、SYK、TJP 2、UCHL 1和WIT-1）。在前列腺上皮细胞系和癌细胞系之间出现差异甲基化和差异调节的大多数基因是新的甲基化靶点，包括PAK 6、RAD 50、TLX 3、PIR 51、MAP 2K 5、INSR、FBN 1和GG 2 -1，代表了用于研究DNA甲基化在前列腺肿瘤中的作用的候选基因的丰富的新来源。
DNA methylation and copy number in the genomes of three immortalized prostate epithelial and five cancer cell lines (LNCaP, PC3, PC3M, PC3M-Pro4, and PC3M-LN4) were compared using a microarray-based technique. Genomic DNA is cut with a methylation-sensitive enzyme HpaII, followed by linker ligation, polymerase chain reaction (PCR) amplification, labeling, and hybridization to an array of promoter sequences. Only those parts of the genomic DNA that have unmethylated restriction sites within a few hundred base pairs generate PCR products detectable on an array. Of 2732 promoter sequences on a test array, 504 (18.5%) showed differential hybridization between immortalized prostate epithelial and cancer cell lines. Among candidate hypermethylated genes in cancer-derived lines, there were eight (CD44, CDKN1A, ESR1, PLAU, RARB, SFN, TNFRSF6, and TSPY) previously observed in prostate cancer and 13 previously known methylation targets in other cancers (ARHI, bc1-2, BRCA1, CDKN2C, GADD45A, MTAP, PGR, SLC26A4, SPARC, SYK, TJP2, UCHL1, and WIT-1). The majority of genes that appear to be both differentially methylated and differentially regulated between prostate epithelial and cancer cell lines are novel methylation targets, including PAK6, RAD50, TLX3, PIR51, MAP2K5, INSR, FBN1, and GG2-1, representing a rich new source of candidate genes used to study the role of DNA methylation in prostate tumors.