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目的:探讨人α-2,6-唾液酸转移酶(ST6GalI)小分子干扰RNA(siRNA)对ST6GalI高表达的结肠癌SW480细胞株的黏附和侵袭力的影响。方法:设计并合成ST6GalI靶向的siRNA,用脂质体Lipofectamine2000包裹后转染SW480细胞。实验设空白对照组、脂质体对照组、非特异性siRNA组和ST6GalIsiRNA组,采用RT-PCR测定细胞中ST6GalImRNA表达水平,流式细胞术检测细胞表面α-2,6-唾液酸含量,并分别用CytoMatrixTM细胞黏附试剂盒及细胞侵袭分析试剂盒,检测细胞对细胞外基质(ECM)黏附与侵袭力。结果:与空白对照组、脂质体对照组、非特异性siRNA组相比,转染48h后,ST6GalIsiRNA组细胞中ST6GalImRNA表达明显下调;转染72h后,ST6GalIsiRNA组细胞表面α-2,6-唾液酸的含量及细胞对ECM的黏附和侵袭力均明显低于其他3个对照组(P<0.05)。结论:化学合成的靶向ST6GalI的siRNA能够下调SW480细胞中ST6GalI基因的表达,降低细胞对ECM的黏附和侵袭力。本实验为进一步研究应用RNA干扰技术抗肿瘤治疗奠定基础。
Objective: To investigate the effect of human α-2,6-sialyltransferase (ST6GalI) siRNA on adhesion and invasion of human colon cancer SW480 cell line with high expression of ST6GalI. Methods: The targeted siRNA of ST6GalI was designed and synthesized and transfected into SW480 cells with Lipofectamine2000. In the experiment, the blank control group, the liposome control group, the nonspecific siRNA group and the ST6GalIsiRNA group were used to detect the expression level of ST6GalI mRNA in the cells by RT-PCR. The cell surface α-2,6-sialic acid content was detected by flow cytometry CytoMatrixTM Cell Adhesion Kit and Cell Invasion Assay Kit were used to detect the adhesion of cells to extracellular matrix (ECM). Results: The expression of ST6GalI mRNA in ST6GalIsiRNA group was significantly decreased 48h after transfection compared with the blank control group, the liposome control group and the non-specific siRNA group. After 72h of transfection, the expression of α-2,6-saliva on the surface of ST6GalIsiRNA group The acid content and cell adhesion and invasiveness of ECM were significantly lower than the other three control groups (P <0.05). Conclusion: Chemically synthesized siRNA targeting ST6GalI can down-regulate the expression of ST6GalI gene in SW480 cells and decrease the adhesion and invasiveness of ECM cells. This experiment lays the foundation for the further study of antitumor therapy using RNA interference technology.