Diallyl disulfide induces ERK phosphorylation and alters gene expression profiles in human colon tumor cells
Diallyl disulfide induces ERK phosphorylation and alters gene expression profiles in human colon tumor cells
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DOI：
10.1093/jn/133.9.2901
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发表时间：
2003-09-01
影响因子：
4.2
通讯作者：
Milner, JA
中科院分区：
文献类型：
--
作者：
Knowles, LM;Milner, JA
二烯丙基二硫化物(DADS)是一种在加工大蒜中发现的化合物，已被证明可以将非同步化的人结肠肿瘤细胞(HCT-15)阻止在细胞周期的G(2)/M期。本研究旨在探讨这种细胞周期阻滞是否与蛋白激酶C(PKC)、钙/钙调素依赖的蛋白激酶II(CAMK II)或细胞外信号调节激酶(ERK)活性的改变有关。DADS(25、50和100Mumol/L)作用于Hct-15细胞4h后，G(2)/M期细胞数分别比对照组增加30%、31%和63%(P<；0.05)。PKC和CAM KII活性不受增加DADS暴露的影响，因此与G(2)/M期细胞阻滞无关。尽管100和500个MUMol/L DADS处理后ERK活性分别增加了44%和60%(P<；0.05)，但接触25或50个MUMol/L DADS对ERK活性没有影响。Western印迹分析显示，尽管DADS(25、50、100和500mumol/L)不影响ERK蛋白的表达，但与对照组相比，其磷酸化程度分别增加了39、52、73和61%(P<；0.05)。这些研究提供的证据表明，ERK信号通路的早期改变可能有助于DADS暴露后观察到的G(2)/M停滞。使用Clonetech Atlas人类癌症基因表达阵列生成的初步数据表明，细胞周期、DNA修复和细胞黏附因子的变化伴随着DADS的暴露，也可能参与调节G(2)/M进展的阻滞。
Diallyl disulfide (DADS), a compound found in processed garlic, has been shown to arrest unsynchronized human colon tumor cells (HCT-15) in the G(2)/M phase of the cell cycle. The present studies were designed to examine whether this cell cycle block related to alterations in protein kinase C (PKC), Ca2+/calmodulin-dependent protein kinase II (CAMK II) or extracellular signal-regulated kinase (ERK) activity. Exposing double thymidine synchronized HCT-1 5 cells to DADS (25, 50 and 100 mumol/L) for 4 h increased the G(2)/M population by 30, 31 and 63%, respectively, compared with controls (P < 0.05). PKC and CAM KII activities were not influenced by increasing DADS exposure and thus did not correlate with the block of cells in the G(2)/M phase. Although ERK activity increased by 44 and 60% after treatment with 100 and 500 mumol/L DADS (P < 0.05), it was not influenced by exposure to 25 or 50 mumol/L DADS. Western blot analysis revealed that although DADS (25, 50, 100 and 500 mumol/L) did not influence the quantity of ERK protein expressed, it did increase its phosphorylation by 39, 52, 73 and 61 %, respectively, compared with controls (P < 0.05). These studies provide evidence that early alterations in ERK pathway signaling may contribute to the G(2)/M arrest observed after DADS exposure. Preliminary data generated using the Clonetech Atlas Human Cancer cDNA Expression Array suggest that alterations in cell cycle, DNA repair and cellular adhesion factors accompany DADS exposure and may also be involved in mediating the block in G(2)/M progression.