Butyrate-induced differentiation of Caco-2 cells is associated with apoptosis and early induction of p21Waf1/Cip1 and p27kip1
Butyrate-induced differentiation of Caco-2 cells is associated with apoptosis and early induction of p21Waf1/Cip1 and p27kip1
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DOI：
10.1067/msy.1998.90365
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发表时间：
1998-08-01
期刊：
影响因子：
3.8
通讯作者：
Townsend, CM
中科院分区：
文献类型：
--
作者：
Litvak, DA;Evers, BM;Townsend, CM
背景。肠粘膜更新是一个增殖、分化和凋亡的过程；其机制在很大程度上仍未明确。本研究的目的是：(1)评估细胞凋亡与肠细胞分化之间的关系，(2)确定细胞周期抑制剂p21（Waf1/Cip1）和p27(Kip1)，细胞凋亡抑制剂Bcl-2和Bcl-X-L是否可能参与其中。肠道源性Caco-2细胞用丁酸钠处理。采用Hoechst染色法、DIVA阶梯法和膜联蛋白V法检测细胞凋亡；以碱性磷酸酶和蔗糖酶活性测定分化程度。分析p21（Waf1/Cip1）、p27（Kip1）和Bcl-2家族成员的RNA和蛋白表达情况。结果：丁酸钠处理Caco-2细胞，可诱导Caco-2细胞分化（碱性磷酸酶和蔗糖酶活性增加）和凋亡。p21（Waf1/Cip1）和p27(KiP1) mRNA和蛋白水平在凋亡或分化前24小时检测到升高；24 h Bcl-2、Bcl-X-L mRNA表达水平下降。Caco-2细胞分化和凋亡同时发生，提示凋亡可能与肠细胞分化有关。诱导p21（Waf1/Cip1）和p27（Kip1）以及下调Bcl-2和Bcl-X-L进一步表明调节肠细胞分化和凋亡的细胞周期机制之间存在联系。
Background. Intestinal mucosal turnover is a process of proliferation, differentiation, and apoptosis; the mechanisms remain largely undefined. The purpose of our study was to (1) assess the relationship between apoptosis and enterocyte differentiation and (2) determine whether the cell-cycle inhibitors, p21(Waf1/Cip1) and p27(Kip1), the apoptosis inhibitors, Bcl-2 and Bcl-X-L, may be involved.Methods. Gut-derived Caco-2 cells were treated with sodium butyrate. Apoptosis was assessed by Hoechst stain, DIVA laddering, and annexin V assay; differentiation was determined by alkaline phosphatase and sucrase activity. RNA and protein were analyzed for expression of p21(Waf1/Cip1), p27(Kip1), and members of the Bcl-2 family.Results, Treatment of Caco-2 cells with sodium butyrate resulted in the concomitant induction of both differentiation (increased alkaline phosphatase and sucrase activity) and apoptosis. Increased level of p21(Waf1/Cip1) and p27(KiP1) mRNA and protein were detected at 24 hours, occurring before apoptosis or differentiation; decreased mRNA level of Bcl-2 and Bcl-X-L were noted at 24 hours.Conclusions. Differentiation and apoptosis occurred simultaneously in Caco-2 cells, suggesting that apoptosis may be linked to enterocyte differentiation. The induction of p21(Waf1/Cip1) and p27(Kip1) and the down-regulation of Bcl-2 and Bcl-X-L further suggest a link between the cell-cycle mechanisms regulating enterocyte differentiation and apoptosis.