Role of fibroblast growth factor receptor signaling in prostate cancer cell survival
Role of fibroblast growth factor receptor signaling in prostate cancer cell survival
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DOI：
10.1093/jnci/93.23.1783
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发表时间：
2001-12-05
期刊：
影响因子：
--
通讯作者：
Ittmann, M
中科院分区：
文献类型：
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作者：
Ozen, M;Giri, D;Ittmann, M
背景：在体内大部分人类前列腺癌和前列腺癌细胞系中，成纤维细胞生长因子（FGF）的表达增加。 FGF 信号传导的改变可能会产生多种影响，包括刺激细胞增殖和抑制细胞死亡。为了确定人类前列腺癌中 FGF 信号传导改变的生物学意义，我们通过在前列腺癌细胞系中表达显性失活 (DN) FGF 受体来破坏信号传导。方法：用 DN FGFR 构建体稳定转染 PC-3、LNCaP 和 DU145 前列腺癌细胞，并用表达 DN FGFR-1 的重组腺病毒感染 LNCaP 和 DU145 细胞。通过集落形成测定、细胞增殖测定、流式细胞术和细胞遗传学分析评估 DN FGFR-1 表达的影响。通过蛋白质印迹法评估细胞周期蛋白 B1 表达，并通过体外激酶测定评估 cdc2 激酶活性，从而评估参与 G(2) 至 M 细胞周期转变的关键调节因子。结果：DN FGFR-1 构建体的稳定转染在所有三种细胞系中抑制集落形成超过 99%。用表达 DN FGFR-1 的腺病毒感染 LNCaP 和 DU145 前列腺癌细胞，导致 48 小时内细胞大量死亡。流式细胞术和细胞遗传学分析表明，DN FGFR-1 受体导致细胞死亡前停滞在细胞周期的 G(2) 期。 Cyclin B1 在 DN FGFR-1 感染的 LNCaP 细胞中积累，但 cdc2 激酶活性降低。结论：这些发现揭示了前列腺癌细胞意外地依赖 FGF 受体信号转导来穿越 G(2)/M 检查点。 G(2) 逮捕的机制尚不清楚。我们的结果提出了 FGF 信号拮抗剂可能增强其他前列腺癌疗法诱导的细胞死亡的可能性。
Background: Expression of fibroblast growth factors (FGFs) is increased in a substantial fraction of human prostate cancers in vivo and in prostate cancer cell lines. Altered FGF signaling can potentially have a variety of effects, including stimulating cell proliferation and inhibiting cell death. To determine the biologic significance of altered FGF signaling in human prostate cancer, we disrupted signaling by expression of a dominant-negative (DN) FGF receptor in prostate cancer cell lines. Methods: PC-3, LNCaP, and DU145 prostate cancer cells were stably transfected with DN FGFR constructs, and LNCaP and DU145 cells were infected with a recombinant adenovirus expressing DN FGFR-1. The effect of DN FGFR-1 expression was assessed by colony-formation assays, cell proliferation assays, flow cytometry, and cytogenetic analysis. Key regulators involved in the G(2)-to-M cell cycle transition were assessed by western blotting to examine cyclin B1 expression and by in vitro kinase assay to assess cdc2 kinase activity. Results: Stable transfection of the DN FGFR-1 construct inhibited colony formation by more than 99% in all three cell lines. Infection of LNCaP and DU145 prostate cancer cells with adenovirus expressing DN FGFR-1 led to extensive cell death within 48 hours. Flow cytometry and cytogenetic analysis revealed that the DN FGFR-1 receptor led to arrest in the G(2) phase of the cell cycle before cell death. Cyclin B1 accumulated in DN FGFR-1-infected LNCaP cells, but cdc2 kinase activity was decreased. Conclusions: These findings reveal an unexpected dependence of prostate cancer cells on FGF receptor signal transduction to traverse the G(2)/M checkpoint. The mechanism for the G(2) arrest is not clear. Our results raise the possibility that FGF-signaling antagonists might enhance the cell death induced by other prostate cancer therapies.