siRNA干扰Ezrin表达对CCL5介导人乳腺癌细胞侵袭能力的影响

目的 探讨Ezrin在趋化因子CCL5促进人乳腺癌细胞MCF-7转移中的作用及其机制.方法 采用小干扰RNA(siRNA)方法下调人乳腺癌细胞中Ezrin的表达,Western blot检测siRNA对Ezrin mRNA和蛋白表达水平下调的变化.干扰后,趋化小室检测MCF-7细胞在不同浓度CCL5作用下的侵袭活性;Western blot检测细胞中总的Ezrin蛋白及磷酸化的Ezrin蛋白表达随CCL5趋化时间的变化.结果 与MCF-7细胞和转染空白质粒pSUPER细胞比较,在CCL5趋化作用下MCF-7细胞Ezrin蛋白的表达无明显增加(P>0.05),干扰后,在50μg/L CCL5作用下,MCF-7细胞表现出侵袭活性最强(F=31.62,P<0.01),但仍较对照组明显降低(F=41.96,P<0.01).结论 趋化因子CCL5促进人乳腺癌细胞MCF-7的趋化与侵袭,Ezrin蛋白的激活在此过程中起着一定的作用.

Role of ezrin in breast cancer cells chemotaxis to CCL5

Objective To study the role of membrane cytoskeleton linker Ezrin in the metastasis of human breast carcinoma cell line MCF-7 facilitated by CC chemokine-5 (CCL5). Methods Two pairs of small interfering RNA (siRNA) targeting Ezrin were transfected into the human breast carcinoma cell line MCF-7cells. Western blotting was used to detect the down-regulation of Ezrin mRNA and protein. The invasion assays were performed on breast cancer cells in the presence of CCL5. Western blotting was used to examine total Ezrin and phosphorylated Ezrin expression in the MCF-7 cells responded to CCL5 at different time points. Results The cells responded chemotactically to CCL5 in a dose-dependent manner (F=31. 62,P< 0.01). The CCL5-induced cell invasion was obviously inhibited by the silencing of ezrin (F=41.96,P<0.01). CCL5 induced a significant up-regulation in the total and the phosphorylated ezrin expression in MCF-7 cells, whereas in the presence of ezrin silencing, the CCL5 induced only a slight increase in the total and the phosphorylated ezrin expression (P>0.05). Conclusion Chemotaxis and invasion of breast cancer cells can be facilitated by CCL5 expression. Our results show that a potential role for Ezrin in the process of the breast cancer cells migration, invasion and metastasis responds to the chemokine CCL5.