摘要
目的 观察不同来源的CC类趋化因子5(CC chemokine ligand 5,CCL5)对人乳腺癌细胞侵袭能力影响并探讨其作用机制.方法 CCL5特异性siRNA慢病毒载体转染人乳腺癌MCF-7细胞,分别用RT-PCR和Western Blot检测细胞CCL5 mRNA和蛋白表达水平,并用细胞侵袭实验检测转染前后细胞侵袭能力的变化.同时,以不同浓度的外源性rhCCL5 (recombinant human CCL5) 作为趋化因素,用细胞侵袭实验检测MCF-7细胞侵袭能力及CC类趋化因子受体5(CCR5)单克隆抗体细胞侵袭能力的影响.结果 CCL5-siRNA慢病毒载体感染可有效降低CCL5 在MCF-7细胞内的表达,细胞的侵袭指数无明显改变,干扰组和阴性对照组侵袭指数差异无统计学意义(P〉0.05).rhCCL5可以明显诱导MCF-7细胞的侵袭(P〈0.05),并且呈浓度依赖趋势;但是这种诱导作用可以部分地被CCR5单克隆抗体阻断,阻断前后细胞的侵袭指数分别为(7.51±0.77)和(3.34±0.51),差异有统计学意义(P〈0.05).结论 细胞内表达的CCL5水平变化对乳腺癌细胞MCF-7的侵袭能力没有影响,而外源性CCL5 可以明显增强MCF-7细胞的侵袭,其机制之一可能是通过与细胞表面的CCR5受体结合.
Objective To examine the effects of the intrinsic and extrinsic CC ehemokine ligand(CCLS) on the invasion of human breast cancer cells MCF-7 and the possible mechanisms. Methods CCL5 specific siRNA was transfected into MCF-7 cells. RT - PCR and Western Blot analysis were used to detect the expression levels of CCL5 in MCF-7 cells. The cell invasion assay was employed to evaluate the invasion ability of the transfected and non - transfected cells. MCF-7 cells ( with or without the treatment of anti - CCR5 antibodies ) were induced by recombinant human CCL5 ( rhCCL5 ) of different concentration and the effect of rhCCL5 on the invasion of MCF-7 cells was detected by using the cell invasion assay as well. Results CCI_5 - specific siRNA transfection could attenuate the expression level of CCL5 in MCF-7 cells effectively(P 〈0.05) ,while the cell invasive ability of these transfected cells were not declined accordingly ( P 〉 0.05 ). rhCCL5 could effectively strengthen the invasion ability of MCF-7 cells in a concentration - dependent manner(P 〈 0.05 ). The inducing effect of rhCCL5 could be partly blocked by anti - CCR5 antibodies and the invasion index was decreased from 7.51 ± 0.77 to 3.34 ±0.51 ( P 〈 0.05). Concimion The intrinsic CCL5 expression level in MCF-7 ceils had not evident effect on the invasive ability of MCF-7 cells,while the extrinsic CCL5 could enhance the invasion of MCF-7 cells effectively.
出处
《临床外科杂志》
2010年第9期600-602,共3页
Journal of Clinical Surgery
基金
湖北省卫生厅资助项目(编号:JS3A14)