Activation of ATM/Chk1 by curcumin causes cell cycle arrest and apoptosis in human pancreatic cancer cells.
Activation of ATM/Chk1 by curcumin causes cell cycle arrest and apoptosis in human pancreatic cancer cells.
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DOI：
10.1038/sj.bjc.6605039
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发表时间：
2009-05-05
影响因子：
8.8
通讯作者：
Srivastava, S. K.
中科院分区：
文献类型：
--
作者：
Sahu, R. P.;Batra, S.;Srivastava, S. K.
姜黄素已被证明可以抑制各种类型的癌细胞的生长;然而，浓度远高于人类临床可达到的水平。人体口服后血浆中姜黄素的浓度估计约为1.8 μM。在这里，我们报告了用2.5 μM姜黄素低剂量单次暴露BxPC-3人胰腺癌细胞24 h，导致细胞显著停滞在G2/M期，并诱导显著的凋亡。免疫印迹研究显示姜黄素处理的细胞中Ser-139处的H2A.X和Ser-280处的Chk 1的磷酸化增加以及DNA聚合酶-β水平降低。H2A.X和Chk 1蛋白的磷酸化是DNA损伤的指标，而DNA聚合酶-β在DNA链断裂的修复中起作用。正常永生化的人胰腺导管上皮（HPDE-6）细胞不受姜黄素处理的影响。此外，我们还观察到Chk 1在Ser-345的磷酸化显著增加，Cdc 25 C在Ser-216的磷酸化显著增加，ATM在Ser-1981的磷酸化轻微增加。姜黄素处理的细胞中，细胞周期蛋白B1和Cdk 1的表达同时减少。此外，姜黄素处理引起BxPC-3细胞中caspase-3和PARP的显著裂解，但在HPDE-6细胞中不引起。通过用ATM或Chk 1特异性siRNA转染BxPC-3细胞来沉默ATM/Chk 1表达，阻断ATM、Chk 1和Cdc 25 C的磷酸化，并保护细胞免于姜黄素介导的G2/M期阻滞和凋亡。这项研究反映了ATM/Chk 1在姜黄素介导的胰腺癌细胞G2/M细胞周期阻滞和细胞凋亡中的关键作用。
Curcumin has been shown to inhibit the growth of various types of cancer cells; however, at concentrations much above the clinically achievable levels in humans. The concentration of curcumin achieved in the plasma after oral administration in humans was estimated to be around 1.8 μM. Here, we report that treatment of BxPC-3 human pancreatic cancer cells with a low and single exposure of 2.5 μM curcumin for 24 h causes significant arrest of cells in the G2/M phase and induces significant apoptosis. Immunoblot studies revealed increased phosphorylation of H2A.X at Ser-139 and Chk1 at Ser-280 and a decrease in DNA polymerase-β level in curcumin-treated cells. Phosphorylation of H2A.X and Chk1 proteins are an indicator of DNA damage whereas DNA polymerase-β plays a role in the repair of DNA strand breaks. Normal immortalised human pancreatic ductal epithelial (HPDE-6) cells remained unaffected by curcumin treatment. In addition, we also observed a significant increase in the phosphorylation of Chk1 at Ser-345, Cdc25C at Ser-216 and a subtle increase in ATM phosphorylation at Ser-1981. Concomitant decrease in the expressions of cyclin B1 and Cdk1 were seen in curcumin-treated cells. Further, curcumin treatment caused significant cleavage of caspase-3 and PARP in BxPC-3 but not in HPDE-6 cells. Silencing ATM/Chk1 expression by transfecting BxPC-3 cells with ATM or Chk1-specific SiRNA blocked the phosphorylation of ATM, Chk1 and Cdc25C and protected the cells from curcumin-mediated G2/M arrest and apoptosis. This study reflects the critical role of ATM/Chk1 in curcumin-mediated G2/M cell cycle arrest and apoptosis in pancreatic cancer cells.
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