沉默黏着斑激酶促进人胃癌SGC-7901细胞骨架蛋白解聚及细胞形态损伤  

FAK silencing promotes the depolymerization of cytoskeleton proteins and morphological damage in SGC-7901 human gastric cancer cells

作　　者：高宁[1] 张静[1] 杜娟[1] 慕明涛[1] 刘俊俊[1] 金文杰[2] 何慧敏[2] 侯颖春[2] 

机构地区：[1]延安大学医学院,延安716000 [2]陕西师范大学生命科学学院,西安710062

出　　处：《中国组织化学与细胞化学杂志》2017年第1期7-12,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助项目(81660492);陕西省高校科协青年人才托举计划(20160223)

摘　　要：目的探讨黏着斑激酶(focal adhesion kinase,FAK)在胃癌细胞骨架结构及形态结构维持中的作用。方法利用靶向FAK的siRNA质粒,转染胃癌SGC-7901细胞,用罗丹明标记的鬼笔环肽及β-tubulin特异性抗体检测细胞骨架微丝及微管蛋白表达;考马斯亮蓝染液对细胞进行染色,观察细胞整体形态结构;环境扫描电子显微镜观察细胞表面结构。结果沉默FAK引起细胞质、细胞伪足及微绒毛等运动相关结构中F-actin解聚。随着沉默时间增长,围绕着细胞核,在细胞质中呈放射状的微管骨架发生解聚,且这种解聚现象呈时间依赖性。FAK沉默还使细胞由梭形、多边形变为圆形,发生脱壁;细胞微绒毛及伪足结构受损,细胞表面趋于光滑。结论 FAK对维持胃癌细胞微丝、微管的分布及癌细胞伪足、微绒毛等恶性形态学特征具有重要作用。Objective To investigate the role of focal adhesion kinase （FAK） in the maintenance of the cytoskeleton and mor- phology of gastric cancer ceils. Methods FAK-specific RNAi plasmids were introduced into gastric cancer cell line SGC-7901 to inhibit FAK expression. Rhodamine labelled phalloidin and 13-tubulin antibodies were used to analyze the proteins composition of microfilament and microtubule. Cell morphology and surface structures were observed by coomassie brilliant blue （CBB） staining and environmental scanning electron microscopy （ESEM）, respectively. Results FAK silencing resulted in the depolymerization of F-actin in the cyto- plasm and motile structures such as pseudopodia and microvilli. With the increase of silencing time, the radial microtubale surrounding cell nuclei in the cytoplasm depolymerized in a time-dependent manner. Moreover, after FAK silencing, the fusiform or polygonal cells detached from culture dish, became round and with smooth surface, the microvilli and pseudopodia were damaged. Conclusion FAK plays an important role in maintaining the distribution of microfilament and microtubule in gastric cancer cells as well as their malignant morphological features such as pseudopodia and microvilli.

关 键 词：粘着斑激酶 RNA干扰 肌动蛋白纤维 微管 细胞骨架 

分 类 号：R362[医药卫生—病理学]