Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell
Chloride Ion Modulates Cell Proliferation of Human Androgen-independent Prostatic Cancer Cell
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DOI：
10.1159/000303042
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发表时间：
2010-01-01
影响因子：
--
通讯作者：
Marunaka, Yoshinori
中科院分区：
文献类型：
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作者：
Hiraoka, Kenji;Miyazaki, Hiroaki;Marunaka, Yoshinori
在本研究中，我们研究了细胞内氯离子是否影响雄激素非依赖性前列腺癌PC3细胞的生长和细胞周期进程。PC3细胞在含113mM C-L的培养液中培养96h，细胞数增加9倍，而在含8 mM-Cl-的培养液中培养96h，细胞生长完全停滞，Rb和cdc2的磷酸化水平降低，Rb和CDc2分别是细胞周期从G(1)期向S期和G(2)期向M期过渡的关键促进剂。在含8 mM-Cl-的培养液中培养细胞，可上调p21(一种CDK抑制剂)的蛋白表达水平，抑制G(1)期向S期的转变，并减少5-乙炔基-2‘-脱氧尿苷(EDU；一种胸苷类似物)的掺入。这些结果表明，低氯培养的细胞延长了细胞周期的所有时相(G(1)、S和G(2)/M)，从而取消了整个细胞周期的进程。低氯条件下培养细胞对细胞周期进程的影响可能是通过改变细胞内氯离子浓度([Cl-](I))来实现的，因为在低氯条件下细胞内[Cl-](I)浓度降低。为了阐明这种可能性，我们研究了速尿和布美他尼这两种Na+/K+/2Cl(-)共转运体(NKCC)抑制剂对PC3细胞增殖的影响。呋塞米和布美他尼可抑制PC3细胞[Cl-](I)和细胞生长。这些结果表明，[Cl-](I)的变化在这种生长机制中起着关键作用。版权所有(C)2010 S.Karger AG，巴塞尔
In the present study, we investigated if the intracellular Cl- affects cell growth and cell cycle progression of androgen-independent prostate cancer PC3 cells. PC3 cells cultured in a medium containing 113 mM C-l- for 96 h grew up 9-fold in cell number, while PC3 cells cultured in an 8 mM-Cl--containing culture medium showed complete arrest of cell growth even after culture for 96 h. Exposure of cells to the 8 mM-Cl- culture medium diminished phosphorylation levels of Rb and cdc2, which are respectively key accelerators of transition from G(1) to S phase and G(2) to M phase in cell cycle progression. Culturing cells in the 8 mM-Cl--containing culture medium upregulated the protein expression level of p21 (a CDK inhibitor) inhibiting transition of G(1) to S phase, and diminished the incorporation of 5-ethynyl-2'-deoxyuridine (EdU; a thymidine analogue) into DNA. These results suggest that cells cultured in the low Cl- medium prolonged the duration of all phases of the cell cycle (G(1), S, and G(2)/M), thereby abolishing overall cell cycle progression. Effects of culturing cells in the low Cl- culture medium on cell cycle progression would be mediated via a change in the intracellular Cl- concentration ([Cl-](i)), since [Cl-](i) was decreased under a low Cl- culture medium. To clarify this possibility, we studied effects of furosemide and bumetanide, Na+/K+/2Cl(-) cotransporter (NKCC) inhibitors, on proliferation of PC3 cells. Furosemide and bumetanide decreased [Cl-](i) and cell growth of PC3 cells. These results suggest that a change in [Cl-](i) would play a critical role in this growth mechanism. Copyright (C) 2010 S. Karger AG, Basel