High-throughput RNAi screening identifies a role for TNK1 in growth and survival of pancreatic cancer cells.
High-throughput RNAi screening identifies a role for TNK1 in growth and survival of pancreatic cancer cells.
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DOI：
10.1158/1541-7786.mcr-10-0436
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Azorsa DO
中科院分区：
文献类型：
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作者：
Henderson MC;Gonzales IM;Arora S;Choudhary A;Trent JM;Von Hoff DD;Mousses S;Azorsa DO
为了鉴定胰腺癌细胞中的新靶点，我们利用高通量RNAi（HT-RNAi）来鉴定当沉默时会降低胰腺癌细胞活力的基因。HT-RNAi筛选涉及用靶向572种激酶的siRNA文库反向转染胰腺癌细胞系BxPC 3。从重复筛选中，约32种激酶被指定为命中，其中22种激酶靶标被选择用于确认和验证。一种被鉴定为来自该筛选的命中的激酶是酪氨酸激酶非受体1（TNK 1），该激酶先前被鉴定为在胚胎干细胞中具有肿瘤抑制剂样特性。用siRNA沉默TNK 1显示在一组胰腺癌细胞系中增殖减少。此外，我们证明了TNK 1的沉默通过半胱天冬酶依赖性途径导致细胞凋亡增加，并且用siRNA靶向TNK 1可以与吉西他滨治疗协同作用。尽管以前的报道表明TNK 1影响Ras和NFκB信号传导，但我们在胰腺癌细胞中没有发现与这些通路类似的相关性。我们的研究结果表明，胰腺癌细胞中的TNK 1不具有胚胎细胞中所见的相同的肿瘤抑制特性，但似乎参与生长和存活。使用HT-RNAi筛选的功能基因组学的应用使我们能够将TNK 1鉴定为胰腺癌细胞中的生长相关激酶。
In order to identify novel targets in pancreatic cancer cells, we utilized high-throughput RNAi (HT-RNAi) to identify genes that, when silenced, would decrease viability of pancreatic cancer cells. The HT-RNAi screen involved reverse transfecting the pancreatic cancer cell line BxPC3 with a siRNA library targeting 572 kinases. From replicate screens, approximately thirty-two kinases were designated as hits, of which twenty-two kinase targets were selected for confirmation and validation. One kinase identified as a hit from this screen was tyrosine kinase non-receptor 1 (TNK1), a kinase previously identified as having tumor suppressor-like properties in embryonic stem cells. Silencing of TNK1 with siRNA showed reduced proliferation in a panel of pancreatic cancer cell lines. Furthermore, we demonstrated that silencing of TNK1 led to increased apoptosis through a caspase-dependent pathway and that targeting TNK1 with siRNA can synergize with gemcitabine treatment. Despite previous reports that TNK1 affects Ras and NFκB signaling, we did not find similar correlations with these pathways in pancreatic cancer cells. Our results suggest that TNK1 in pancreatic cancer cells does not possess the same tumor suppressor properties seen in embryonic cells, but appears to be involved in growth and survival. The application of functional genomics using HT-RNAi screens has allowed us to identify TNK1 as a growth-associated kinase in pancreatic cancer cells.