摘要
目的探讨不同迁移特性骨肉瘤亚克隆细胞系粘着斑激酶(FAK)的表达及其对骨肉瘤细胞增殖和迁移的影响。方法通过有限稀释法获得不同迁移能力的骨肉瘤细胞143B亚克隆细胞系A1、A2、A3、A4、A5,运用Western blottting技术检测不同亚克隆细胞系FAK的表达。小干扰RNA(siRNA)序列研究FAK对骨肉瘤细胞增殖及迁移能力的作用,免疫荧光染色法比较形成板状伪足细胞数的比例。结果骨肉瘤细胞143B亚克隆细胞系A2、A3的迁移能力较A1、A4和A5强(P<0.05),其细胞中FAK表达也明显增高。外源转入FAK siRNA后,骨肉瘤亚克隆细胞系A3的迁移能力明显下降(P<0.05),形成板状伪足细胞数的比例由33.3%降至12.8%(P<0.05),但增殖能力无明显变化。结论迁移能力较强的骨肉瘤细胞143B亚克隆细胞系中FAK呈高表达,FAK可能通过影响板状伪足的形成从而增强骨肉瘤细胞的迁移能力,但是对骨肉瘤细胞的增殖无明显作用。
Objective To explore the expression of focal adhesion kinase（FAK） in different migration characteristics of os- teosarcoma subclone cell lines, as well as its role in the osteosarcoma cell proliferation and migration. Methods Limiting dilution was used to obtain 143B subclone cell lines（ A1, A2, A3, A4, A5 ）with different migration capacities, and their FAK expressions were detec- ted by Western blottting. The role of FAK in the osteosarcoma cells proliferation and migration was studied by small interfering RNA （siRNA） sequence,and the proportion of the ceils with lamellipodia was counted by immunofluorescence staining method. Results The migration ability of osteosarcoma cell lines 143B subclone A2 and A3 was significantly stronger than A1, A4 and A5（P 〈0. 05）, and FAK expressions of cell lines A2 and A3 were significantly higher. The subclone cell line A3 transfected with FAK-siRNA de- creased the migration ability（P 〈0. 05） ,and the proportion of cells with lamellipodia decreased from 33.3% to 12. 8% （P 〈0. 05）, but there was no change of the cell growth. Conclusion FAK was founded in the 143B osteosarcoma subclone cell lines with stronger migration capacities. FAK may affect the migration ability of osteosarcoma cells through lamellipodia formation, but have no effect on the proliferation of osteosarcoma cells.
出处
《临床肿瘤学杂志》
CAS
2013年第5期385-389,共5页
Chinese Clinical Oncology
基金
江苏省自然科学基金面上资助项目(BK2012775)
关键词
骨肉瘤
亚克隆细胞系
FAK
增殖
迁移
板状伪足
Osteosarcoma
Subclone cell lines
Focal adhesion kinase（FAK）
Proliferation
Migration
Lamellipodia
