针对layilin的shRNA抑制透明质酸诱导人肺腺癌A549细胞体外黏附和侵袭

Inhibitory effects of shRNA targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma A549 cells induced by hyaluronan in vitro

背景与目的透明质酸参与了多种肿瘤细胞黏附侵袭行为。layilin是一种新发现的与细胞骨架有密切联系的透明质酸受体。本研究的目的是观察针对layilin的短发夹RNA(shorthairpinRNA,shRNA)对体外培养的人肺腺癌A549细胞layilin表达及受透明质酸诱导黏附侵袭行为的影响。方法设计和构建带有U6启动子的能产生针对layilin的shRNA的RNA干扰质粒,转染至A549细胞,以RT-PCR、Westernblot和免疫荧光检测A549细胞转染前后layilin表达,以平板黏附模型和Boyden小室模型检测A549细胞转染前后黏附和侵袭能力。结果和转染前相比,转染了RNA干扰表达质粒的A549细胞layilin表达明显减少(P<0.01),而且受透明质酸诱导而黏附于平板和穿过Boyden小室隔膜的A549细胞数皆明显减少(P<0.01)。结论针对layilin的shRNA能明显抑制透明质酸诱导的人肺腺癌A549细胞体外黏附和侵袭能力。

Background and objective It has been proved that hyaluronan was involved in adhesion and invasion behavior of varied tumor cells. Layilin is a receptor of byaluronan found recently and has close relationship with cytoskeleton. The aim of this study is to investigate the effect of short hairpin RNA （sbRNA） targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma cell line A549 induced by byaluronan in vitro. Methods RNA interference plasmid that included U6 promoter and could express sbRNA 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 byaluronan also significantly decreased （P〈0. 01）. Conclusion The sbRNA targeting layilin can efficiently inhibit the adhesive and invasive ability of A549 cells induced by hyaluronan in vitro.