A novel approach to identify driver genes involved in androgen-independent prostate cancer.
A novel approach to identify driver genes involved in androgen-independent prostate cancer.
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DOI：
10.1186/1476-4598-13-120
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发表时间：
2014-05-23
期刊：
影响因子：
37.3
通讯作者：
Trobridge GD
中科院分区：
文献类型：
--
作者：
Schinke EN;Bii V;Nalla A;Rae DT;Tedrick L;Meadows GG;Trobridge GD
插入突变筛选已成功地用于鉴定癌基因和肿瘤抑制基因。通常，这些筛选使用伽玛逆转录病毒(γRV)或转座子作为插入突变剂。然而，在肿瘤发生过程中较晚发生的具有复制能力的γRV或转座子的插入突变可以产生不会推动癌症进展的乘客突变。在这里，我们利用复制能力不强的慢病毒载体(LV)进行插入突变筛选，以确定雄激素非依赖性前列腺癌(AIPC)进展过程中的基因。用LV诱变前列腺癌细胞，以在载体整合位点附近由失调基因(S)提供的雄激素缺乏环境中丰富具有选择性优势的克隆。我们使用体外AIPC模型和体内AIPC异种移植模型进行筛查。我们的方法利用穿梭载体确定了前病毒整合位点，该载体允许在大肠杆菌中快速拯救含有LV长末端重复序列(LTR)-染色体连接的质粒。这种穿梭载体方法不需要聚合酶链式反应扩增，与基于聚合酶链式反应的技术相比有几个优点。在雄激素缺乏的培养液(p <； 0.001)中生长的细胞中，前列腺癌易感基因附近的前病毒整合被丰富，并鉴定出五个影响AIPC的候选基因：ATPAF1、GCOM1、MEX3D、PTRF和TRPM4。此外，我们还发现，在雄激素缺乏的条件下，ATPAF1的RNAi敲除显著降低了生长(p <； 0.05)。我们的方法已经被证明用于前列腺癌，识别出一个已知的前列腺癌基因PTRF，以及几个以前与前列腺癌无关的基因。无复制能力的穿梭载体方法在癌症基因发现以及询问不同的生物学和疾病过程方面具有广泛的潜在应用。
Insertional mutagenesis screens have been used with great success to identify oncogenes and tumor suppressor genes. Typically, these screens use gammaretroviruses (γRV) or transposons as insertional mutagens. However, insertional mutations from replication-competent γRVs or transposons that occur later during oncogenesis can produce passenger mutations that do not drive cancer progression. Here, we utilized a replication-incompetent lentiviral vector (LV) to perform an insertional mutagenesis screen to identify genes in the progression to androgen-independent prostate cancer (AIPC). Prostate cancer cells were mutagenized with a LV to enrich for clones with a selective advantage in an androgen-deficient environment provided by a dysregulated gene(s) near the vector integration site. We performed our screen using an in vitro AIPC model and also an in vivo xenotransplant model for AIPC. Our approach identified proviral integration sites utilizing a shuttle vector that allows for rapid rescue of plasmids in E. coli that contain LV long terminal repeat (LTR)-chromosome junctions. This shuttle vector approach does not require PCR amplification and has several advantages over PCR-based techniques. Proviral integrations were enriched near prostate cancer susceptibility loci in cells grown in androgen-deficient medium (p < 0.001), and five candidate genes that influence AIPC were identified; ATPAF1, GCOM1, MEX3D, PTRF, and TRPM4. Additionally, we showed that RNAi knockdown of ATPAF1 significantly reduces growth (p < 0.05) in androgen-deficient conditions. Our approach has proven effective for use in PCa, identifying a known prostate cancer gene, PTRF, and also several genes not previously associated with prostate cancer. The replication-incompetent shuttle vector approach has broad potential applications for cancer gene discovery, and for interrogating diverse biological and disease processes.
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影响因子：
30.8
作者：
通讯作者：
--
将转录因子识别为单一的同源物2作为前列腺癌中潜在的生物标志物和免疫疗法靶标。
DOI：
10.1158/1078-0432.ccr-09-0911
发表时间：
2009-09-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Arredouani MS;Lu B;Bhasin M;Eljanne M;Yue W;Mosquera JM;Bubley GJ;Li V;Rubin MA;Libermann TA;Sanda MG
通讯作者：
Sanda MG
影响因子：
2.8
作者：
Gould, M. L.;Williams, G.;Nicholson, H. D.
通讯作者：
Nicholson, H. D.
影响因子：
4.8
作者：
Lu, S;Tsai, SY;Tsai, MJ
通讯作者：
Tsai, MJ
影响因子：
3.7
作者：
Le TP;Sun M;Luo X;Kraus WL;Greene GL
通讯作者：
Greene GL