[靶向层粘连蛋白的短发夹RNA对透明质酸诱导的人肺腺癌A549细胞体外黏附及侵袭行为的抑制作用]
[Inhibitory effects of shRNA targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma A549 cells induced by hyaluronan in vitro].
作者信息
Zhuo Wenlei, Wang Yan, Chen Zhengtang
机构信息
Institute of Cancer, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037, P.R.China.
出版信息
Zhongguo Fei Ai Za Zhi. 2006 Jun 20;9(3):236-40. doi: 10.3779/j.issn.1009-3419.2006.03.05.
BACKGROUND
It has been proved that hyaluronan was involved in adhesion and invasion behavior of varied tumor cells. Layilin is a receptor of hyaluronan found recently and has close relationship with cytoskeleton. The aim of this study is to investigate the effect of short hairpin RNA (shRNA) targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma cell line A549 induced by hyaluronan in vitro.
METHODS
RNA interference plasmid that included U6 promoter and could express shRNA targeting layilin was designed, constructed, and transfected into A549 cell line. Layilin expression was examined by RT-PCR, Western blot and immunofluorescence. Adhesive and invasive ability was examined by plate adhesion model and Boyden chamber model.
RESULTS
After plasmid transfected, layilin expression in A549 cells obviously decreased (P < 0.01), and the numbers of adhesive A549 cells on plate and A549 cells permeating septum of Boyden chamber induced by hyaluronan also significantly decreased (P < 0.01).
CONCLUSIONS
The shRNA targeting layilin can efficiently inhibit the adhesive and invasive ability of A549 cells induced by hyaluronan in vitro.
背景
已证实透明质酸参与多种肿瘤细胞的黏附和侵袭行为。层黏蛋白受体是最近发现的透明质酸受体，与细胞骨架关系密切。本研究旨在探讨靶向层黏蛋白受体的短发夹RNA（shRNA）对体外透明质酸诱导的人肺腺癌细胞系A549黏附和侵袭行为的影响。
方法
设计、构建包含U6启动子并能表达靶向层黏蛋白受体shRNA的RNA干扰质粒，转染至A549细胞系。通过RT-PCR、Western印迹和免疫荧光检测层黏蛋白受体的表达。采用平板黏附模型和Boyden小室模型检测黏附及侵袭能力。
结果
质粒转染后，A549细胞中层黏蛋白受体表达明显降低（P < 0.01），透明质酸诱导的平板上黏附的A549细胞数量以及穿过Boyden小室隔膜的A549细胞数量也显著减少（P < 0.01）。
结论
靶向层黏蛋白受体的shRNA能有效抑制体外透明质酸诱导的A549细胞的黏附和侵袭能力。
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