β1-整合素反义寡核苷酸抑制胃癌细胞系SGC7901粘附侵袭的实验研究

Inhibition of Cell Attachment and Invasion of Human Gastric Cancer Cell line SGC7901 by Integrinβ1 asODN

  • 摘要: 目的:观察β1-整合素反义寡核苷酸对高侵袭性人胃癌细胞SGC7901粘附侵袭的抑制作用。方法:以脂质体为载体将硫代磷酸修饰的反义寡核苷酸转染SGC7901细胞,免疫化学方法观察转染后以Biotin标记的Integrinβ1 asODN在细胞内的分布;RT-PCR及流式细胞术分别测定Integrinβ1 mRNA和蛋白表达变化;MTT法和Boy-den小室模型分别测定其粘附力和侵袭力。结果:Integrinβ1 asODN转染SGC7901后1h,胞浆及胞核内可见均匀分布的浅棕色颗粒,随着时间的延长,颗粒颜色不断加深;RT-PCR结果显示489bp处条带不同程度变浅;Integrinβ1蛋白表达曲线明显左移;粘附和侵袭实验证明,Integrinβ1 asODN转染的SGC7901细胞的粘附力及侵袭力明显下降(P<0.001),抑制率分别为54%和76%,明显优于随机序列(P<0.001)。结论:Integrinβ1 asODN转染SGC7901可降低其mRNA和蛋白表达水平,从而抑制其对细胞外基质(ECM)的粘附和侵袭。

     

    Abstract: Objective : To observe the in hibitio n of adhesion and invasion of SGC7901 to ECM by Integrinβ1 asODN. Methods : ODN were transfected into SGC7901, taking liposome as vector. The distributio n of ODN was recorded by im munochem istry, changes in the expression of Integrinβ1 mRNA and protein were determined by RT-P C R and FCM, respectively. The adhesion and invasion to ECM were measured by MTT and Boyden chamber method,respectively. Results : A fter Integrinβ1 asODN was transfected into SGC7901 it distributed evenly in cytoplasm and nucleus. PCR and FCM revealed a weakened band on 489bp and a shift-left curve,respectively. Adhesion and invasion assay also presented a decrease,with the in hibitin g rate of 54% and 76% . The extent of decrease induced by Integrinβ1 asODN was larger than that by the random ODN (P<0.001). Conclusion : Transfection of Integ rinβ1 asODN into SGc/901 induced a decrease in the expression of Integrinβ1 m RNA and protein, thus a decrease in adhesion and invasion to ECM , with a better effect than random ODN.

     

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