ST6Gal I基因特异性siRNA对SW480细胞黏附和侵袭力的影响

EffectofST6GalIsiRNA-mediatedgene silencing on the adhesion and invasion of SW480 cells

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。

AIM： To study the effect of synthesized ST6Gal Ⅰ specific siRNA on the adhesion and invasiveness of human colon carcinoma cell line SW480 with over expression of ST6Gal Ⅰ. METHODS： A double strand small interference RNA （siRNA） targeting ST6Gal Ⅰ was designed and synthesized, and then transfected into SW480 cells by lipofectmine 2000. SW480 cells were cultured and divided into 4 groups： blank control group, liposome control group, nonspecific siRNA group and ST6Gal Ⅰ siRNA group. The expression of ST6Gal Ⅰ mRNA was examined by RT-PCR and the amount of α-2, 6-sialylation on the SW480 cell surface was detected by flow cytometry. The adhesion and invasion of SW480 cells to extracellular matrix （ECM） were analyzed by using CytoMatrix^TM kit and cell invasion assay kit, respectively. RESULTS： After SW480 cells were transfected for 48 hours, the expression of ST6Gal Ⅰ mRNA in ST6Gal Ⅰ siRNA group was significantly decreased compared with that in the blank control group, liposome control group, and nonspecific siRNA group （ P 〈 0. 05）. After SW480 cells were transfected for 72 hours, the amount of α-2,6-sialylation on cell surface, the adhesion and invasion of the cells in ST6Gal Ⅰ siRNA group were markedly lower than those in the other 3 groups （ P 〈 0. 05）. CONCLUSION： The chemically synthesized specific siRNA tar qetincl ST6Gal Ⅰ can effectively inhibit the expression of ST6Gal Ⅰ and reduce cell adhesion and invasion to ECM in SW480 cells. Our research is important for further study of anti-tumor treatment with RNA interference.