Small interfering RNA targeting integrin-linked kinase inhibited the growth and induced apoptosis in human bladder cancer cells
Small interfering RNA targeting integrin-linked kinase inhibited the growth and induced apoptosis in human bladder cancer cells
复制标题
靶向整合素连接激酶的小干扰RNA抑制人膀胱癌细胞的生长并诱导细胞凋亡
DOI：
10.1016/j.biocel.2011.05.003
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发表时间：
2011-09-01
影响因子：
4
通讯作者：
Chen, Jun-Xia
中科院分区：
文献类型：
--
作者：
Gao, Juan;Zhu, Jun;Chen, Jun-Xia
整合素连接激酶（ILK）是一种细胞内丝氨酸/苏氨酸激酶，与细胞生长和存活、细胞周期进展、肿瘤血管生成和细胞凋亡有关。近年来的研究表明，ILK在多种实体瘤中的表达和活性显著增强。然而，ILK在肿瘤中的确切分子机制尚未完全确定。本研究的目的是利用基于质粒载体的小干扰RNA（siRNA）来确定ILK的敲低是否会抑制膀胱癌细胞的生长并诱导细胞凋亡。实验结果表明，ILK基因的敲除可显著抑制膀胱癌BIU-87和EJ细胞的增殖和生长，调节细胞周期，诱导细胞凋亡。我们通过Western印迹和免疫荧光分析证明，ILK的敲低抑制了下游信号传导靶蛋白激酶B/Akt、糖原合成酶激酶3-β（GSK-3 β）的磷酸化，并降低了BIU-87和EJ细胞中β-连环蛋白的表达。此外，ILK的下调还可增加核糖核酸酶抑制因子（RI）的表达，RI是一种重要的具有多种功能的酸性胞质蛋白。转染ILK siRNA的BIU-87细胞裸鼠皮下注射，肿瘤生长受到明显抑制，肿瘤重量减轻，微血管密度降低，细胞凋亡率升高。结论：ILK可能参与膀胱癌的发生发展，并可能成为膀胱癌治疗的新靶点。我们的研究可能具有生物学和临床意义。(c)2011爱思唯尔有限公司保留所有权利。
Integrin-linked kinase (ILK), an intracellular serine/threonine kinase, is implicated in cell growth and survival, cell-cycle progression, tumor angiogenesis, and cell apoptosis. Recent studies showed that the expression and activity of ILK increased significantly in many types of solid tumors. However, the exact molecular mechanism of ILK underlie tumor has not been fully ascertained. The purpose of our study was to determine whether knockdown of ILK would inhibit cell growth and induce apoptosis in bladder cancer cells using a plasmid vector based small interfering RNA (siRNA). The experiments showed that knockdown of ILK could remarkably inhibit cell proliferation and growth, regulate cell cycle and induce apoptosis of bladder cancer BIU-87 and EJ cells. We demonstrated that knockdown of ILK inhibited phosphorylation of downstream signaling targets protein kinase B/Akt, glycogen synthase kinase 3-beta (GSK-3 beta), and reduced expression of beta-catenin in BIU-87 as well as EJ cells by Western blot and Immunofluorescence analysis. In addition, down-regulation of ILK also could increase expression of Ribonuclease inhibitor (RI), an important acidic cytoplasmic protein with many functions. BALB/C nude mice injected with the BIU-87 cells transfected ILK siRNA showed a significant inhibition of the tumor growth with lighter tumor weight, lower microvessels density and higher apoptosis rate than those in the other two control groups. In conclusion, these results suggest that ILK might be involved in the development of bladder cancer, and could be served as a novel potential therapy target for human bladder cancer. Our study may be of biological and clinical importance. (c) 2011 Elsevier Ltd. All rights reserved.