Integrated, genome-wide screening for hypomethylated oncogenes in salivary gland adenoid cystic carcinoma.
Integrated, genome-wide screening for hypomethylated oncogenes in salivary gland adenoid cystic carcinoma.
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DOI：
10.1158/1078-0432.ccr-10-2992
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发表时间：
2011-07-01
期刊：
影响因子：
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通讯作者：
Ha PK
中科院分区：
文献类型：
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作者：
Shao C;Sun W;Tan M;Glazer CA;Bhan S;Zhong X;Fakhry C;Sharma R;Westra WH;Hoque MO;Moskaluk CA;Sidransky D;Califano JA;Ha PK
涎腺腺样囊性癌（ACC）是一种罕见的恶性肿瘤，是知之甚少。为了寻找启动子甲基化控制下的相关癌基因候选者，进行了整合的全基因组筛选。用5-氮杂-2 ′-脱氧胞苷（5-Aza dC）和曲古抑菌素A（TSA）对正常涎腺细胞株进行整体去甲基化，然后进行表达阵列分析。ACC特异性表达谱是使用表达微阵列分析的主要ACC和正常样品。接下来，整合这两个图谱以鉴定基因的子集，用于进一步验证ACC与正常ACC中的启动子去甲基化。最后，在较大的ACC队列中进一步验证了有希望的候选者的mRNA、蛋白质和启动子甲基化水平。然后在癌细胞系中进行功能验证。我们发现159个基因在5-Aza dC/TSA处理后显著重新表达，并在ACC中过表达。经过初步验证，8个候选基因在ACC中显示低甲基化：AQP 1，CECR 1，C1 QR 1，CTAG 2，P53 AIP 1，TDRD 12，BEX 1和DYNLT 3。水通道蛋白1（AQP 1）表现出最显着的低甲基化，并进一步验证。两个独立的队列证实了ACC中AQP 1的低甲基化。值得注意的是，在石蜡包埋的ACC队列中，AQP 1的mRNA和蛋白质均显著过表达。此外，AQP 1在5-Aza dC/TSA处理的SACC 83中上调。最后，AQP 1促进SACC 83细胞增殖和集落形成。我们的综合，全基因组筛选方法被证明是一种有效的策略，检测新的癌基因在ACC。AQP 1是一个有前途的癌基因候选ACC和转录调控启动子低甲基化。
Salivary gland adenoid cystic carcinoma (ACC) is a rare malignancy that is poorly understood. In order to look for relevant oncogene candidates under the control of promoter methylation, an integrated, genome-wide screen was performed. Global demethylation of normal salivary gland cell strains using 5-aza-2′-deoxycytidine (5-Aza dC) and Trichostatin A (TSA), followed by expression array analysis was performed. ACC-specific expression profiling was generated using expression microarray analysis of primary ACC and normal samples. Next, the two profiles were integrated to identify a subset of genes for further validation of promoter demethylation in ACC versus normal. Finally, promising candidates were further validated for mRNA, protein, and promoter methylation levels in larger ACC cohorts. Functional validation was then performed in cancer cell lines. We found 159 genes that were significantly re-expressed after 5-Aza dC/TSA treatment and overexpressed in ACC. After initial validation, eight candidates showed hypomethylation in ACC: AQP1, CECR1, C1QR1, CTAG2, P53AIP1, TDRD12, BEX1, and DYNLT3. Aquaporin 1 (AQP1) showed the most significant hypomethylation and was further validated. AQP1 hypomethylation in ACC was confirmed with two independent cohorts. Of note, there was significant overexpression of AQP1 in both mRNA and protein in the paraffin-embedded ACC cohort. Furthermore, AQP1 was up-regulated in 5-Aza dC/TSA treated SACC83. Lastly, AQP1 promoted cell proliferation and colony formation in SACC83. Our integrated, genome-wide screening method proved to be an effective strategy for detecting novel oncogenes in ACC. AQP1 is a promising oncogene candidate for ACC and is transcriptionally regulated by promoter hypomethylation.