摘要
目的:研究在裸鼠结肠癌肝转移模型中,富含脯氨酸的蛋白酪氨酸激酶2(proline-rich tyrosine kinase 2,Pyk2)对人结肠癌细胞肝转移能力和超微结构的影响。方法:利用正义及反义cDNA技术,在人结肠癌细胞系SW480基础上,构建Pyk2高表达的细胞系SW480-Pyk2+及低表达的细胞系SW480-Pyk2-作为实验组,并以转染空载体的细胞系SW480-vector和原细胞系SW480作为对照组。4组共32只4周龄的Babl/c nu/nu雄性裸鼠,用脾注射保留脾脏法构建裸鼠肝转移瘤模型。6周后处死裸鼠,用Western blot检测瘤细胞中Pyk2蛋白的表达,计数肝转移瘤数目,电子显微镜下观察肝转移瘤细胞的超微结构。结果:6周后裸鼠肝转移模型建立成功,Western blot显示SW480-Pyk2+组的蛋白表达量明显高于空载体组(SW480-vector)和SW480组,SW480-Pyk2-组的蛋白表达量明显低于SW480组和SW480-vector组;肝转移瘤计数显示SW480-Pyk2+组明显少于SW480-vector组及SW480组,SW480-Pyk2-组明显多于SW480-vector组及SW480组(P<0.05),而SW480-vector组与SW480组之间无明显差异;电子显微镜下观察发现,SW480-Pyk2+组细胞连接发育较成熟,有形成腺腔趋势,微绒毛较多,细胞器较多,粗面内质网较多,细胞异型性较轻;而SW480-Pyk2-组细胞连接发育不良,细胞器少,微绒毛短小而稀疏,细胞核异形明显,有核沟。结论:Pyk2能减少结肠癌肝转移灶的数量,改善细胞的异型性和细胞连接发育,并可能通过此机制抑制肿瘤转移。
Objective：To study the influence of proline-rich tyrosine kinase 2（Pyk2） on hepatic metastasis and microstructure of colon cancer cells in nude mice.Methods： Hepatic metastases of colonic cancer were established in Babl/c nu/nu nude mice by splenic injection of colonic cancer cells SW480 which were transfected with sense-Pyk2 plasmid and antisense-Pyk2 plasmid.After six weeks,we counted the number of liver metastases in each group and observed the ultrastructure of tumor cells in each group by electron microscopy;furthermore,we tried to find out the correlationship between Pyk2 expression and the number of colonic cancer liver metastases.Results： Difference was found between the nude mice injected with sense-Pyk2 plasmid and antisense-Pyk2 plasmid,and there was correlation between Pyk2 expression and the number of liver metastases,Pyk2 could inhabit the liver metastasis of colonic adenocarcinoma.In contrast,the ultrastructure of tumor cell had less heteromorphism in nude mice transfected with sense-Pyk2 plamisd.Conclusion： Pyk2 can reduce the number of liver metastasis of colonic adenocarcinoma,and also the heteromorphism of tumor cells which inhabit the progress of colonic adenocarcinoma liver metastasis.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2011年第2期204-208,共5页
Journal of Peking University:Health Sciences
基金
国家自然科学基金资助(30572105)~~
关键词
黏着斑激酶2
结肠肿瘤
肝转移
超微结构
Focal adhesion kinase 2； Colonic neoplasms； Hepatic metastasis； Ultrastructure
