TRPV2 Activation Induces Apoptotic Cell Death in Human T24 Bladder Cancer Cells: A Potential Therapeutic Target for Bladder Cancer
TRPV2 Activation Induces Apoptotic Cell Death in Human T24 Bladder Cancer Cells: A Potential Therapeutic Target for Bladder Cancer
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DOI：
10.1016/j.urology.2010.03.029
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发表时间：
2010-08-01
期刊：
影响因子：
2.1
通讯作者：
Shimada, Shoichi
中科院分区：
文献类型：
--
作者：
Yamada, Takahiro;Ueda, Takashi;Shimada, Shoichi
目的 探讨瞬时受体电位香草酸 2 (TRPV2) 通道蛋白在人尿路上皮癌 (UC) 细胞中的功能表达，并确定钙离子通过 TRPV2 流入 UC 细胞是否参与细胞凋亡。 材料与方法 分析 TRPV2 mRNA 在膀胱癌细胞系（低分化 UC 细胞系 T24 和高分化 UC 细胞系 RT4）中的表达。逆转录酶-聚合酶链式反应。使用钙成像测定法研究 T24 细胞中 TRPV2 通道的钙渗透性，该测定使用大麻二酚 (CBD)（一种相对选择性的 TRPV2 激动剂）和钌红 (RuR)（一种非选择性 TRPV 通道拮抗剂）。使用细胞活力测定评估 CBD 存在下 T24 或 RT4 细胞的死亡。使用膜联蛋白-V 测定和 TRPV2 的小干扰 RNA (siRNA) 沉默证实了 CBD 引起的 T24 细胞凋亡。结果 TRPV2 mRNA 在 T24 细胞中大量表达。 UC细胞中的表达水平与高度疾病相关。 CBD 的施用增加了 T24 细胞的细胞内钙浓度。此外，随着 CBD 浓度的增加，T24 细胞的活力逐渐降低，而 RT4 细胞大多不受影响。细胞死亡是通过钙通过TRPV2持续流入引起的细胞凋亡而发生。结论UC细胞中的TRPV2通道是钙渗透性的，通过这些通道调节钙流入直接导致UC细胞死亡。 UC 细胞中的 TRPV2 通道可能是一个潜在的新治疗靶点，特别是在更高级别的 UC 细胞中。泌尿学 76：509.e1-509.e7，2010。(C) 2010 Elsevier Inc.
BJECTIVES To investigate the functional expression of the transient receptor potential vanilloid 2 (TRPV2) channel protein in human urothelial carcinoma (UC) cells and to determine whether calcium influx into UC cells through TRPV2 is involved in apoptotic cell death.MATERIAL AND METHODS The expression of TRPV2 mRNA in bladder cancer cell lines (T24, a poorly differentiated UC cell line and RT4, a well-differentiated UC cell line) was analyzed using reverse transcriptase-polymerase chain reaction. The calcium permeability of TRPV2 channels in T24 cells was investigated using a calcium imaging assay that used cannabidiol (CBD), a relatively selective TRPV2 agonist, and ruthenium red (RuR), a nonselective TRPV channel antagonist. The death of T24 or RT4 cells in the presence of CBD was evaluated using a cellular viability assay. Apoptosis of T24 cells caused by CBD was confirmed using an annexin-V assay and small interfering RNA (siRNA) silencing of TRPV2.RESULTS TRPV2 mRNA was abundantly expressed in T24 cells. The expression level in UC cells was correlated with high-grade disease. The administration of CBD increased intracellular calcium concentrations in T24 cells. In addition, the viability of T24 cells progressively decreased with increasing concentrations of CBD, whereas RT4 cells were mostly unaffected. Cell death occurred via apoptosis caused by continuous influx of calcium through TRPV2.CONCLUSIONS TRPV2 channels in UC cells are calcium-permeable and the regulation of calcium influx through these channels leads directly to the death of UC cells. TRPV2 channels in UC cells may be a potential new therapeutic target, especially in higher-grade UC cells. UROLOGY 76: 509.e1-509.e7, 2010. (C) 2010 Elsevier Inc.