膜联蛋白A5与宫颈黏膜上皮细胞癌变的关系

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。

Relationship between annexin A5 and carcinogenesis of uterine cervical cells

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 NaHCO_3.Total cell RNA was obtained by Trizol method.The expression of annexin A5 was tested by RT-PCR,Western blot and immu- nohistochemical 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 immunohisto- chemical 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.