比较CasKi和H8细胞中心体γ.doc

比较CasKi和H8细胞中心体γ-tubulin 和Aurora-A 激酶表达的差异 巫静娴 赵涌 （重庆医科大学分子医学与肿瘤研究中心，病理教研室，重庆 400016） 摘 要：目的 对比分析HPV16 E6E7阳性的人宫颈癌细胞（CasKi）和人宫颈鳞状上皮永生化细胞（H8）中γ-tubulin和中心体相关激酶Aurora-A表达的差异。方法 采用间接免疫荧光显示γ-tubulin的荧光强度来分析CasKi和H8细胞中心体的差异， RT-PCR检测中心体相关激酶Aurora-A mRNA水平， Western blot方法对γ-tubulin、Aurora-A在两种细胞中的表达水平进行半定量分析。结果 与H8细胞相比，CasKi细胞中心体γ-tubulin荧光信号更强，γ-tubulin、Aurora-A mRNA和蛋白表达更高，差异具有统计学意义（p0.05）。结论 宫颈癌细胞CasKi较癌前的永生化细胞H8的中心体蛋白过度表达、中心体异常更显著，提示中心体结构和功能的异常可能是宫颈癌早期形成中的关键步骤。 【关键词】 宫颈癌，永生化，诺帝，中心体，γ-tubulin，Aurora-A 【中图号】R737.3 【文献标识码】A Comparation the expreesion diffences of γ-tubulin and Aurora-A in human cervical cancer cell CasKi and H8 cells Wu Jingxian , Zhao Yong (Molecular Medicine and Cancer Research Centre Department of pathology , Chongqing Medical University , Chongqing 400016,China ) Abstract: Objective To compare and analyze the expression differences of γ-tubulin and Aurora-A in human cervical cancer cell CasKi and immortalized human cervical cell H8 with positive HPV16 E6E7. Methods Indirect immuno- fluorescence was used to show the fluorescence intensity of γ-tubulin to analyze the centrosome difference of Caski and H8. RT-PCR was applied to detect the Aurora-A mRNA level and Western blot was used to semi-quantitatively analyze the expressions of γ-tubulin and Aurora-A in two cells. Results Compared with H8 cells, the fluorescence signal of γ-tubulin in Caski cells was stronger, meanwhile the expression of γ-tubulin and Aurora-A were higher. Conclusions Centrosome of Caski was over-expressed compared with that of H8 and the centrosome abnormality was more prominent, which suggested that the abnormality of centrosome structure and function may be the key to the early formation of cervical carcinoma. Key words: cervical carcinoma, immortalization, centrosome, γ-tubulin, Aurora-A 中心体异常是人类肿瘤中的常见现象，包括中心体数目的扩增、体积的肥大或中心体蛋白的异常磷酸化及其功能特性的改变。随后越来越多的报道观察到中心体在多种恶性肿瘤组织中的异常改变，如乳腺癌[1]、头颈鳞癌[2]、神经外胚层肿瘤[3]等等。而在宫颈癌前病变及其进展中探讨中心体扩增情况，目前研究领域却鲜有涉及。本实验旨在通过检测宫颈癌细胞CasK