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为了研究β-半乳糖α2,6-唾液酸转移酶1(ST6Gal-I)对人绒毛膜癌细胞(JAR)和人绒毛膜外滋养层细胞(HTR-8/SVneo)迁移能力的影响。首先采用Transwell小室的实验方法比较两种细胞的迁移能力,再用PCR和Western blot的方法检测ST6Gal-I mRNA及其蛋白的表达差异,最后通过Transwell检测经siRNA干扰ST6Gal-I的表达后对细胞迁移能力的影响,并通过免疫共沉淀技术检测ST6Gal-I的靶蛋白整合素β1唾液酸化程度的变化。结果表明,JAR细胞的迁移能力强于HTR-8/SVneo细胞;JAR细胞中ST6Gal-I在mRNA及蛋白水平表达均高于HTR-8/SVneo细胞,同时检测到JAR细胞中靶蛋白整合素β1的唾液酸化程度亦高于对照组。用siRNA干扰相对高表达的JAR细胞中ST6Gal-I基因,发现其迁移能力随之下降,且ST6Gal-I的靶蛋白整合素β1的唾液酸化程度也发生降低。表明ST6Gal-I参与了肿瘤细胞的迁移调控。该研究结果对发现新的治疗靶点有重要的启示。
To investigate the effect of beta-galactosyl-2,6-sialyltransferase 1 (ST6Gal-I) on the migration of human choriocarcinoma cells (JAR) and human chorionic trophoblast cells (HTR-8 / SVneo) Transwell chamber was used to compare the migration ability of two kinds of cells. PCR and Western blot were used to detect the expression of ST6Gal-I mRNA and protein. Finally, the expression of ST6Gal-I was detected by Transwell assay. Migration ability and the co-immunoprecipitation technique was used to detect the changes of sialylation of integrin β1, the target protein of ST6Gal-I. The results showed that the migration ability of JAR cells was stronger than that of HTR-8 / SVneo cells. The expression of ST6Gal-I mRNA and protein in JAR cells was higher than that in HTR-8 / SVneo cells and JAR cells target protein integrinβ1 Sialylation level is also higher than the control group. Using siRNA to interfere with the ST6Gal-I gene in relatively highly expressed JAR cells, the migration ability was found to decrease, and the degree of sialylation of integrin β1, the target protein of ST6Gal-I, also decreased. ST6Gal-I is involved in the regulation of tumor cell migration. The results of this study have important implications for the discovery of new therapeutic targets.