肝癌细胞生物学行为改变涉及p120ctn酪氨酸磷酸化.docVIP

肝癌细胞生物学行为改变涉及p120ctn酪氨酸磷酸化( 黄华艺 农朝赞 郭凌宵 （广西民族医院 530001） [摘 要] 目的 观察p120ctn磷酸化与肝癌细胞生物学行为的关系，探讨p120ctn肝癌细胞粘附和信号转导中的作用。方法 使用免疫沉淀和免疫印迹法检测EGF引起的p120ctn磷酸化情况，用免疫细胞化学技术观察p120ctn在细胞内的分布、转位和形态学改变，用相应的方法检测细胞粘附和迁移情况，反义核酸技术用于反义p120ctn的转染。结果 EGF可使p120ctn酪氨酸发生磷酸化，以作用20min时最明显；磷酸化后的p120ctn发生了转位，膜表达减少，核表达增加，(-catenin也发生相似的改变；细胞粘附能力下降，迁移能力增加，细胞形态变为细长和不规则，伪足增多。结论 p120ctn在细胞粘附和酪氨酸蛋白激酶信号转导途径中起重要作用。提示p120ctn酪氨酸磷酸化与肝癌发生和转移过程存在某种联系。 [关键词] 酪氨酸磷酸化 肝癌 连环蛋白p120 P120ctn Tyrosine Phosphorylation Are Involved in Hepatocellular Carcinoma Cell Biologic Behavior Changes ? HUANG Hua-yi1*, NONG Chao-zan1, GUO Ling-xiao1,ZHA Xi-liang2§. Guangxi Nationalities Hospital, Guangxi Nanning 530001,China 【Abstract】Objective In order to investigate the relationship between p120ctn phosphorylation and of biologic behaviors liver cancer cells for further exploring the role of p120ctn in hepatocellular carcinoma cell adhesion and signaling. Methods Phosphotyrosine of p120ctn were detected by immunoprecipitation and immunoblotting. The cellular distribution and translocation of p120ctn and (-catenin and cell morphology changes were detected and examined by immunocytochemistry.Cell adhesion and migration abilities were also detected using correspondent methods. Antiseuse-nucleotide method were used in anti-p120ctn transfection. Results Phosphorylated tyrosine of p120ctn had been observed after EGF stimulation,The tyrosine phosphorylation level was significant at 20min after EGF stimulation comparied to control. Phosphorylated p120ctn translocated into nucleus, its membrane distribution was reduced, and same changes of (-catenin were found while compared with p120ctn; Cell adhesion abilities reduced and migration abilities increased, cell morphology became thin, elongated and irregular, their speudopods increased. Conclusion p120ctn play an important role in hepatocellular carcinoma cell adhesion and protein tyrosine kinase signaling cascade. It suggested that there is a relationship between p120ctn tyrosin