Gasi Delila, Trapman Jan
Department of Pathology, Josephine Nefkens Institute, Erasmus University Medical Centre, Rotterdam, The Netherlands.
Methods Mol Biol. 2011;776:335-48. doi: 10.1007/978-1-61779-243-4_19.
Fusion between androgen-regulated TMPRSS2 and ETS transcription factor gene ERG is the most frequent genetic alteration that occurs in 40-70% of prostate cancers. Not only ERG but also other ETS transcription factor genes are involved in gene fusions. ETV1, ETV4, and ETV5 have all several fusion partners. One common feature shared by the majority of these partners is androgen-regulated expression. Despite its high frequency, the biological and molecular effects of ETS gene fusion in prostate cancer development and progression are unknown. In this chapter quantitative polymerase chain reaction (Q-PCR) is used for detection and further studying the incidence and properties of these fusion transcripts. The focus is on the expression of TMPRSS2-ERG transcripts in clinical prostate samples. Androgen regulation of TMPRSS2 is measured in commonly used LNCaP prostate cancer cells grown with and without the synthetic androgen R1881. Furthermore, combining Q-PCR with 5' RLM-RACE and sequencing are described for the identification of novel ETS fusion partners.
雄激素调节的跨膜丝氨酸蛋白酶2（TMPRSS2）与ETS转录因子基因ERG之间的融合是前列腺癌中最常见的基因改变，在40%-70%的前列腺癌中发生。不仅ERG，其他ETS转录因子基因也参与基因融合。ETV1、ETV4和ETV5都有多个融合伴侣。这些伴侣中的大多数共有的一个共同特征是雄激素调节的表达。尽管其频率很高，但ETS基因融合在前列腺癌发生和发展中的生物学和分子效应尚不清楚。在本章中，定量聚合酶链反应（Q-PCR）用于检测并进一步研究这些融合转录本的发生率和特性。重点是临床前列腺样本中TMPRSS2-ERG转录本的表达。在添加和不添加合成雄激素R1881的情况下培养常用的LNCaP前列腺癌细胞，检测TMPRSS2的雄激素调节。此外，还描述了将Q-PCR与5' RLM-RACE和测序相结合以鉴定新型ETS融合伴侣的方法。
