Su Jian-Min, Wang Li-Ying, Liang Yu-Long, Zha Xi-Liang
Department of Biochemistry and Molecular Biology, Shanghai Medical College of Fudan University, Shanghai, China.
World J Gastroenterol. 2005 Aug 14;11(30):4667-73. doi: 10.3748/wjg.v11.i30.4667.
Cell adhesion molecules and their signal molecules play a very important role in carcinogenesis. The aim of this study is to elucidate the role of these molecules and the signal molecules of integrins and E-cadherins, such as (focal adhesion kinase) FAK, (integrin linked kinase) ILK, and beta-catenin in hepatocellular carcinoma cell apoptosis.
We first synthesized the small molecular compound, S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and identified it, by element analysis and (1)H NMR. To establish the apoptosis model of the SMMC-7721 hepatocellular carcinoma cell, we treated cells with DCVC in EBSS for different concentrations or for various length times in the presence of 20 micromol/L N,N'-diphenyl-p-phenylenediamine, which blocks necrotic cell death and identified this model by flow cytometry and DNA ladder. Then we studied the changes of FAK, ILK, beta-catenin, and PKB in this apoptotic model by Western blot.
We found that the loss or decrease of cell adhesion signal molecules is an important reason in apoptosis of SMMC-7721 hepatocellular carcinoma cell and the apoptosis of SMMC-7721 cell was preceded by the loss or decrease of FAK, ILK, PKB, and beta-catenin or the damage of cell-matrix and cell-cell adhesion.
Our results suggested that the decrease of adhesion signal molecules, FAK, ILK, PKB, and beta-catenin, could induce hepatocellular carcinoma cell apoptosis.
细胞黏附分子及其信号分子在肿瘤发生过程中发挥着非常重要的作用。本研究旨在阐明这些分子以及整合素和E-钙黏蛋白的信号分子，如黏着斑激酶（FAK）、整合素连接激酶（ILK）和β-连环蛋白在肝癌细胞凋亡中的作用。
我们首先合成了小分子化合物S-(1,2-二氯乙烯基)-L-半胱氨酸（DCVC），并通过元素分析和氢核磁共振（1H NMR）对其进行鉴定。为建立SMMC-7721肝癌细胞凋亡模型，我们在含有20微摩尔/升N,N'-二苯基对苯二胺的Earle's平衡盐溶液（EBSS）中，用不同浓度或不同时长的DCVC处理细胞，该溶液可阻断坏死性细胞死亡，并通过流式细胞术和DNA梯状条带鉴定该模型。然后我们通过蛋白质免疫印迹法研究该凋亡模型中FAK、ILK、β-连环蛋白和蛋白激酶B（PKB）的变化。
我们发现细胞黏附信号分子的缺失或减少是SMMC-7721肝癌细胞凋亡的重要原因，且SMMC-7721细胞凋亡之前存在FAK、ILK、PKB和β-连环蛋白的缺失或减少，或细胞-基质及细胞-细胞黏附的损伤。
我们的结果表明，黏附信号分子FAK、ILK、PKB和β-连环蛋白的减少可诱导肝癌细胞凋亡。
