趋化因子受体7小干扰RNA对人头颈部鳞状细胞癌细胞增殖及侵袭的影响

目的 研究趋化因子受体7(chemokine receptor7,CCR-7)小干扰RNA(small interfering RNA,siRNA)高表达的人头颈部鳞状细胞癌(squamous cell carcinoma of head and neck,SCCHN)细胞系PCI-4B增殖和侵袭能力的影响,探讨siRNA技术在SCCHN基因治疗中的可行性和特异性.方法 选用阳离子脂质体LipofectamineTM 2000作为转染试剂,将针对人CCR-7基因的siRNA转染PCI-4B,采用Western blotting法检测CCR-7蛋白水平,应用甲基噻唑基四唑(MTT)法检测转染前后细胞增殖变化,应用激光共聚焦显微镜观察细胞的改变,Transwell趋化小室法检测细胞体外侵袭能力的改变,并通过荧光显微镜及TransAMTM细胞核因子西乙蛋白(nuclear favor-kappa B,NF-Κb)p65活性检测试剂盒检测NF-Κb细胞内定位及活性.结果 CCR-7 siRNA转染后,细胞内CCR-7蛋白表达水平显著降低(P＜0.05);CCL-19诱导使SCCHN细胞增殖能力增高,细胞极性浓聚、迁移和侵袭能力增高、NF-Κb活化及核转位,这些效应均被CCR-7 siRNA抑制.结论 CCR-7 siRNA在体外成功地下调CCR-7的蛋白水平,抑制了CCL-19增强PCI-4B增殖能力和迁移能力的效应,为临床SCCHN的治疗提供了新方法.

Effect of chemokine receptor 7 small interfering RNA on proliferation and invasion of squamous cell carcinoma of head and neck

Objective To study the effect of chemokine receptor 7(CCR-7)small interfering RNA (siRNA)on proliferation and invagion of squamous cell carcinoma of head and neck(SCCHN).Methods CCR-7 siRNA was co-transfected into SCCHN cell line PCI-4B using LipofectamineTM 2000.CCR-7 protein level was detected by western blotting.SCCHN cell proliferation was detected by MTT,and the change of actin cytoskeleton observed by confocal laser scanning microscope.Transwell assays were used to determine chemotaxis and invasion of SCCHN cells.The activity and nuclear translocation of nuclear factor-kappa B (NF-κB)were detected by TransAMTM NF-κB p65 kit and fluorescence microscope respectively.Results After CCR-7 siRNA transfection.the protein level of CCR-7 was significantly decreased.The changes induced by CCL-19,including increased proliferation rate,polarized actin polymerization,increased chemotaxis rate and invasion rate,were all abolished by CCR-7 siRNA transfection.CCR-7 siRNA also diminished CCL-19-induced NF-κB activation and nuclear translocation.Conclusions CCR-7 siRNA could inhibit expression of CCR-7 and diminish the increased proliferation and invasion of SCCHN induced by CCL-19 in vitro.CCR-7 siRNA may provide a potential treatment strategy for SCCHN.