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目的对比人乳头瘤病毒(human papillomavirus,HPV)16E6E7基因阳性的人宫颈癌细胞(CasKi细胞株)和人宫颈鳞状上皮永生化细胞(H8细胞株)中γ-tubulin和中心体相关激酶Aurora-A表达的差异并探讨其意义。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-AmRNA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。结果宫颈癌细胞CasKi中γ-tubulin的免疫荧光信号(57.78±3.13)明显强于H8细胞(37.37±2.37)(P<0.05);Aurora-AmRNA在CasKi细胞(0.745±0.013)中的表达显著高于H8细胞(0.617±0.223)(P<0.05);Westernblot结果显示Aurora-A和γ-tubulin在CasKi细胞中的蛋白表达水平分别为(0.969±0.011),(0.909±0.019)明显高于H8细胞[分别为(0.568±0.016),(0.571±0.022),P<0.05]。结论中心体的异常扩增和过表达可能是宫颈癌前病变转变成癌的早期生物学标志,中心体异常可能成为诊断宫颈上皮内瘤变进展为癌的辅助分子生物学指标。
Objective To compare the expression of γ-tubulin and centrosome-associated kinase Aurora-1 in human cervical carcinoma (CasKi) cells and human cervical squamous cell immortalized cells (H8) cells with human papillomavirus (HPV) 16E6E7 gene, A expression differences and explore its significance. Methods The fluorescence intensity of γ-tubulin was detected by indirect immunofluorescence to analyze the difference of centrosomes between CasKi and H8 cells. The level of Aurora-AmRNA was determined by RT-PCR. The expression of γ-tubulin and Aurora-A in two The expression levels in the cells were subjected to semi-quantitative analysis. Results The immunofluorescence signal of γ-tubulin in CasKi cervical carcinoma cells (57.78 ± 3.13) was significantly stronger than that in H8 cells (37.37 ± 2.37) (P <0.05). The expression of Aurora-AmRNA in CasKi cells (0.745 ± 0.013) (0.617 ± 0.223) in H8 cells (P <0.05). The results of Western blot showed that the protein expression levels of Aurora-A and γ-tubulin in CasKi cells were (0.969 ± 0.011) and (0.909 ± 0.019) [(0.568 ± 0.016) and (0.571 ± 0.022), respectively, P <0.05]. Conclusions Abnormal amplification and overexpression of centrosome may be the early biological markers of cervical precancerous lesions turning into carcinomas. The centrosome abnormality may become an auxiliary molecular biological indicator for the diagnosis of cervical intraepithelial neoplasia.