中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2009年
1期
5-10
,共6页
癌%鳞状细胞%受体%趋化因子%肿瘤浸润
癌%鱗狀細胞%受體%趨化因子%腫瘤浸潤
암%린상세포%수체%추화인자%종류침윤
Carcinoma,squamous cell%Receptors,chemokine%Neoplasm invasiveness
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.
目的 研究趨化因子受體7(chemokine receptor7,CCR-7)小榦擾RNA(small interfering RNA,siRNA)高錶達的人頭頸部鱗狀細胞癌(squamous cell carcinoma of head and neck,SCCHN)細胞繫PCI-4B增殖和侵襲能力的影響,探討siRNA技術在SCCHN基因治療中的可行性和特異性.方法 選用暘離子脂質體LipofectamineTM 2000作為轉染試劑,將針對人CCR-7基因的siRNA轉染PCI-4B,採用Western blotting法檢測CCR-7蛋白水平,應用甲基噻唑基四唑(MTT)法檢測轉染前後細胞增殖變化,應用激光共聚焦顯微鏡觀察細胞的改變,Transwell趨化小室法檢測細胞體外侵襲能力的改變,併通過熒光顯微鏡及TransAMTM細胞覈因子西乙蛋白(nuclear favor-kappa B,NF-Κb)p65活性檢測試劑盒檢測NF-Κb細胞內定位及活性.結果 CCR-7 siRNA轉染後,細胞內CCR-7蛋白錶達水平顯著降低(P＜0.05);CCL-19誘導使SCCHN細胞增殖能力增高,細胞極性濃聚、遷移和侵襲能力增高、NF-Κb活化及覈轉位,這些效應均被CCR-7 siRNA抑製.結論 CCR-7 siRNA在體外成功地下調CCR-7的蛋白水平,抑製瞭CCL-19增彊PCI-4B增殖能力和遷移能力的效應,為臨床SCCHN的治療提供瞭新方法.
목적 연구추화인자수체7(chemokine receptor7,CCR-7)소간우RNA(small interfering RNA,siRNA)고표체적인두경부린상세포암(squamous cell carcinoma of head and neck,SCCHN)세포계PCI-4B증식화침습능력적영향,탐토siRNA기술재SCCHN기인치료중적가행성화특이성.방법 선용양리자지질체LipofectamineTM 2000작위전염시제,장침대인CCR-7기인적siRNA전염PCI-4B,채용Western blotting법검측CCR-7단백수평,응용갑기새서기사서(MTT)법검측전염전후세포증식변화,응용격광공취초현미경관찰세포적개변,Transwell추화소실법검측세포체외침습능력적개변,병통과형광현미경급TransAMTM세포핵인자서을단백(nuclear favor-kappa B,NF-Κb)p65활성검측시제합검측NF-Κb세포내정위급활성.결과 CCR-7 siRNA전염후,세포내CCR-7단백표체수평현저강저(P＜0.05);CCL-19유도사SCCHN세포증식능력증고,세포겁성농취、천이화침습능력증고、NF-Κb활화급핵전위,저사효응균피CCR-7 siRNA억제.결론 CCR-7 siRNA재체외성공지하조CCR-7적단백수평,억제료CCL-19증강PCI-4B증식능력화천이능력적효응,위림상SCCHN적치료제공료신방법.
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.
