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.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2006年第4期433-435,共3页
Chinese Journal of Anatomy
