The TREK2 Channel Is Involved in the Proliferation of 253J Cell, a Human Bladder Carcinoma Cell
The TREK2 Channel Is Involved in the Proliferation of 253J Cell, a Human Bladder Carcinoma Cell
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DOI：
10.4196/kjpp.2013.17.6.511
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发表时间：
2013-12-01
影响因子：
2
通讯作者：
Kim, Yangmi
中科院分区：
文献类型：
--
作者：
Park, Kyung-Sun;Han, Min Ho;Kim, Yangmi
膀胱癌是吸烟男性中第七大常见癌症，发病率随着年龄的增长而增加。发生机制尚未确定。钾离子通道与细胞增殖有关。一些双孔结构域K+通道（K2 P），如TASK 3和TREK 1，最近已被证明在癌细胞中过表达。在这里，我们专注于细胞生长和机械敏感性K2 P通道，TREK 2，在人膀胱癌细胞系，253 J之间的关系。我们通过Western blot和实时荧光定量PCR证实了TREK 2在膀胱癌细胞系中的表达。使用膜片钳技术，在对称的150 mM KCl溶液的存在下记录机械敏感性TREK 2通道。在253 J细胞中，TREK 2通道被多不饱和脂肪酸、-60 mV的细胞内酸中毒和-40 mV或40 mV的机械牵拉激活。此外，与阴性对照siRNA相比，小干扰RNA（siRNA）介导的TREK 2敲低导致从-19.9 mV +/- 0.8（n=116）至-8.5 mV +/- 1.4（n=74）的轻微去极化和253 J细胞的增殖降低。TREK 2 siRNA处理的253 J细胞显示细胞周期边界蛋白p21和p53的表达显著增加，并且细胞周期蛋白D1和D3的蛋白表达也显著降低。总之，TREK 2通道存在于膀胱癌细胞系中，并且可能至少部分地有助于细胞周期依赖性生长。
Bladder cancer is the seventh most common cancer in men that smoke, and the incidence of disease increases with age. The mechanism of occurrence has not yet been established. Potassium channels have been linked with cell proliferation. Some two-pore domain K+ channels (K2P), such as TASK3 and TREK1, have recently been shown to be overexpressed in cancer cells. Here we focused on the relationship between cell growth and the mechanosensitive K2P channel, TREK2, in the human bladder cancer cell line, 253J. We confirmed that TREK2 was expressed in bladder cancer cell lines by Western blot and quantitative real-time PCR. Using the patch-clamp technique, the mechanosensitive TREK2 channel was recorded in the presence of symmetrical 150 mM KCl solutions. In 253J cells, the TREK2 channel was activated by polyunsaturated fatty acids, intracellular acidosis at -60 mV and mechanical stretch at -40 mV or 40 mV. Furthermore, small interfering RNA (siRNA)-mediated TREK2 knockdown resulted in a slight depolarization from -19.9 mV +/- 0.8 (n=116) to -8.5 mV +/- 1.4 (n=74) and decreased proliferation of 253J cells, compared to negative control siRNA. 253J cells treated with TREK2 siRNA showed a significant increase in the expression of cell cycle boundary proteins p21 and p53 and also a remarkable decrease in protein expression of cyclins D1 and D3. Taken together, the TREK2 channel is present in bladder cancer cell lines and may, at least in part, contribute to cell cycle-dependent growth.