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作 者:李欣[1] 高福禄[2] 何晓文[3] 王芳[3] 孙树汉[3]
机构地区:[1]河北医科大学,石家庄050017 [2]承德医学院组织学与胚胎学教研室,承德067000 [3]第二军医大学遗传学教研室,上海200433
出 处:《解剖学杂志》2006年第4期433-435,共3页Chinese Journal of Anatomy
摘 要:目的:确定在宫颈癌细胞系Hela细胞和SiHa细胞中是否有膜联蛋白A5(anxA5)的表达,为研究anxA5的功能及其与宫颈癌的关系奠定基础。方法:培养Hela细胞和SiHa细胞。TRIzol法提取细胞总RNA,RT- PCR法检测anxA5在mRNA水平的表达并将其基因片段连接于T载体,测序仪测序;Western印迹法和免疫细胞化学ABC法检测anxA5的表达。结果:两种宫颈癌细胞的RT-PCR结果和Western印迹法均显示目的条带;免疫细胞化学染色可见胞质和核膜被染成棕黄色;测序并经NCBI的BLAST进行比对,显示为人anxA5。结论:anxA5在宫颈癌细胞系Hela细胞和SiHa细胞的mRNA水平和蛋白质水平均有表达。Objective: To confirm whether annexin A5 is expressed in the uterine cervical cell line SiHa cells and Hela cells, providing a base for studying the relationship between annexin A5 and uterine cervical carcinoma. Methods: The cells were cultured in DMEM supplemented with 10% FBS and 2 g/L NaHCO3. Total cell RNA was obtained by Trizol method. The expression of annexin A5 was tested by RT-PCR, Western blot and immunohistochemical staining. Annexin A5 obtained was linked on T-vector for sequencing. Results: Both RT-PCR and Western blot showed the objective band. The cytoplasm of the cells were stained yellow by immunohistochemical staining. Sequencing showed the gene was human annexin A5. Conclusion: Both the uterine cervical cell lines SiHa and Hela express annexin A5 at both the mRNA and the protein levels.
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