P27(Kip1), regulated by glycogen synthase kinase-3β, results in HMBA-induced differentiation of human gastric cancer cells.
P27(Kip1), regulated by glycogen synthase kinase-3β, results in HMBA-induced differentiation of human gastric cancer cells.
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DOI：
10.1186/1471-2407-11-109
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发表时间：
2011-03-27
期刊：
影响因子：
3.8
通讯作者：
Gu Q
中科院分区：
文献类型：
--
作者：
Wei M;Wang Z;Yao H;Yang Z;Zhang Q;Liu B;Yu Y;Su L;Zhu Z;Gu Q
胃癌是全球癌症相关死亡率的第二大常见原因。虽然去分化预测胃癌预后不良，但去分化的分子机制尚未阐明，这可能为肿瘤的发生和发展提供基本的见解。此外，最近发现的分化诱导剂异丙二乙酰胺（HMBA）的作用的分子机制需要调查，目前还没有关于HMBA对胃癌的作用的研究报道。基于FACS分析的结果，在HMBA和LiCl的单次处理和顺序组合后，使用蛋白质印迹法测定参与细胞周期或凋亡的蛋白质的水平。转染Ad-Akt后上调Akt表达，Western blotting检测GSK-3β和质子泵的表达。采用激酶活性测定、免疫沉淀和蛋白质印迹法研究HMBA对GSK-3β、CDK 2和CDK 4蛋白定位和活性的影响。此外，进行北方印迹和RNA酶保护测定以确定HMBA的功能浓度。HMBA增加p27 Kip 1表达并诱导胃上皮细胞分化相关的细胞周期阻滞。此外，用HMBA处理胃衍生细胞诱导G 0/G1期阻滞和质子泵上调，质子泵是胃癌分化的标志。此外，HMBA处理增加GSK-3β在细胞核中的表达和活性，但不增加细胞质中的GSK-3β的表达和活性。HMBA降低CDK 2活性，诱导p27 Kip 1表达，GSK-3β抑制剂可使其恢复。此外，HMBA增加了p27 Kip 1与CDK 2的结合，并且这种结合被GSK-3β抑制剂消除。本研究结果提示GSK-3β通过调节p27 Kip 1与CDK 2的组装发挥作用，从而在HMBA诱导的胃上皮细胞分化相关的G 0/G1阻滞中发挥关键作用。
Gastric cancer is the second most common cause of global cancer-related mortality. Although dedifferentiation predicts poor prognosis in gastric cancer, the molecular mechanism underlying dedifferentiation, which could provide fundamental insights into tumor development and progression, has yet to be elucidated. Furthermore, the molecular mechanism underlying the effects of hexamethylene bisacetamide (HMBA), a recently discovered differentiation inducer, requires investigation and there are no reported studies concerning the effect of HMBA on gastric cancer. Based on the results of FACS analysis, the levels of proteins involved in the cell cycle or apoptosis were determined using western blotting after single treatments and sequential combinations of HMBA and LiCl. GSK-3β and proton pump were investigated by western blotting after up-regulating Akt expression by Ad-Akt infection. To investigate the effects of HMBA on protein localization and the activities of GSK-3β, CDK2 and CDK4, kinase assays, immunoprecipitation and western blotting were performed. In addition, northern blotting and RNase protection assays were carried out to determine the functional concentration of HMBA. HMBA increased p27Kip1 expression and induced cell cycle arrest associated with gastric epithelial cell differentiation. In addition, treating gastric-derived cells with HMBA induced G0/G1 arrest and up-regulation of the proton pump, a marker of gastric cancer differentiation. Moreover, treatment with HMBA increased the expression and activity of GSK-3β in the nucleus but not the cytosol. HMBA decreased CDK2 activity and induced p27Kip1 expression, which could be rescued by inhibition of GSK-3β. Furthermore, HMBA increased p27Kip1 binding to CDK2, and this was abolished by GSK-3β inhibition. The results presented herein suggest that GSK-3β functions by regulating p27Kip1 assembly with CDK2, thereby playing a critical role in G0/G1 arrest associated with HMBA-induced gastric epithelial cell differentiation.
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影响因子：
64.5
作者：
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