caveolin-1脚手架样结构域多肽对HEp2细胞迁移和侵袭能力的影响  

Effects of caveolin-scaffolding domain polypeptide on migration and invasion ability of HEp2 cell

作　　者：顾栋桦[1] 付琳琳[1] 平金良[1] 

机构地区：[1]浙江省湖州市中心医院病理科,浙江湖州313000

出　　处：《中国现代医生》2016年第30期29-32,共4页China Modern Doctor

基　　金：浙江省科技厅公益类项目(2014C33245);浙江省湖州市科技计划项目(2013GYB11)

摘　　要：目的研究caveolin-1脚手架样结构域(caveolin-scaffolding domain,CSD)多肽对HEp2细胞迁移和侵袭能力的影响,并探讨其分子机制。方法 CSD多肽和乱序多肽由人工合成得到,并在N'末端用生物素标记。用免疫细胞化学法检测CSD多肽在HEp2细胞中的渗透情况,划痕实验检测细胞的迁移能力,Transwell细胞体外侵袭实验检测细胞的侵袭能力;免疫印迹法检测HEp2细胞中FAK蛋白的表达水平及磷酸化程度。结果免疫细胞化学检测显示CSD多肽能够较好地进入HEp2细胞内;CSD多肽组HEp2细胞迁移能力明显低于乱序组和空白对照组(P均<0.01),而乱序肽组与空白对照组之间差异无统计学意义(P>0.05);CSD多肽组发生侵袭的细胞数明显低于乱序组和空白对照组(P均<0.01),而乱序肽组与空白对照组之间差异无统计学意义(P>0.05);免疫印迹法检测结果显示,CSD多肽组FAK相对磷酸化程度明显低于乱序组与空白对照组(P均<0.01)。结论 caveolin-1脚手架样结构域多肽可顺利进入HEp2细胞内,而且可以抑制肿瘤细胞的迁移和侵袭能力,FAK磷酸化水平的下调可能是其中重要的分子机制。Objective To study the effects of caveolin-scaffolding domain （CSD） polypeptide on migration and invasion ability of HEp2, and to explore its molecular mechanism. Methods CSD polypeptide and out-of-order polypeptide were synthesized artificially, and was marked by biotin at N＇-end. Immunocytochemical method was applied to test the infil- tration condition of CSD polypeptide in HEp2 cells, scratch test was applied to test the migration ability of.the cells, and Transwell in vitro invasion assay was applied to test the invasion ability of the cells; Western blot method was ap- plied to test the expression~!evel and phosphorylation degree of FAK protein in HEp2 cells. Results According to im- munocytochemical test, it was shown that CSD polypeptide was able to enter HEp2 ceils favorably; the migration ability of HEp2 cells in CSD polypeptide group was significantly lower than those in out-of-order polypeptide group and blank controI group（P〈0.01）, but there was no significant difference between out-of-order polypeptide group and blank control group（P〉0.05）; the number of cells with invasion in CSD polypeptide group was significantly lower than those in out- of-order polypeptide group and blank control group（P〈0.01）, but there was no significani difference between out-of-order polypeptide group and blank control group （P〉0.05）; according to Western blot method, it was shown that relative phosphorylation degree of FAK in CSD polypeptide group was significantly lower than those in out-of-order polypeptide group and blank control group（P〈0.01）. Conclusion Caveolin-1 scaffolding domain polypeptide is able to successfully enter HEp2 cells, and is also able to inhibit the migration and invasion ability of tumor cells： The down-regulating of FAK phosphorylation level may be one of the important molecular mechanisms.

关 键 词：CAVEOLIN-1 粘着斑激酶 多肽 鳞状细胞癌 肿瘤浸润 

分 类 号：Q73-3[生物学—分子生物学]