皮层肌动蛋白通过与Src相互作用促进食管鳞状细胞癌的侵袭和转移  

Cortactin accelerates invasion and migration of esophageal squamous carcinoma cells through interaction with Src

作　　者：张九娜[1] 翟山[2] 杨阳 郭永泽[1] 李校天[1] ZHANG Jiuna;ZHAI Shan;YANG Yang;GUO Yongze;LI Xiaotian(Department of Gastroenterology,Affiliated Hospital of Hebei University of Engineering,Handan 056002,Hebei,China)

机构地区：[1]河北工程大学附属医院消化内科,河北邯郸056002 [2]河北工程大学附属医院病理科,河北邯郸056002 [3]冀中能源邢矿总医院内四科

出　　处：《上海医学》2021年第12期907-912,共6页Shanghai Medical Journal

基　　金：河北省科技计划项目(16277742D)。

摘　　要：目的探讨皮层肌动蛋白(cortactin,CTTN)对食管鳞状细胞癌(简称鳞癌)迁移侵袭的作用及分子机制。方法在人食管鳞癌细胞系EC109、TE1中通过慢病毒转染技术过表达或敲减CTTN;蛋白质印迹法检测细胞间黏附分子-1(ICAM1)、基质金属蛋白酶(MMP)2、MMP9、血管内皮生长因子C(VEGF-C)的表达量;通过Transwell侵袭实验评估细胞侵袭力,划痕试验观察细胞迁移能力;应用免疫共沉淀验证与CTTN相互作用的蛋白质。结果在EC109和TE1细胞中成功过表达或敲减CTTN,并利用嘌呤霉素筛选出稳定的细胞。与对照组相比,CTTN过表达组的ICAM1、MMP2、MMP9、VEGF-C表达量显著增高(P值均<0.01);同时,过表达组的细胞侵袭转移能力较对照组也显著增强[穿膜细胞数(541.0±17.8)个比(230.8±10.9)个,划痕愈合速度(72.8±1.9)%比(42.8±1.6)%,P值均<0.001]。CTTN敲减组与对照组相比,ICAM1、MMP2、MMP9、VEGF-C表达量显著降低,细胞侵袭转移能力也显著降低[穿膜细胞数(102.3±4.0)个比(223.8±12.1)个,划痕愈合速度(25.7±1.5)%比(43.5±1.9)%,P值均<0.001]。免疫共沉淀实验证实CTTN与Src存在直接相互作用。结论CTTN通过与Src相互作用促进食管鳞癌细胞的侵袭和转移。Objective To investigate the effects and mechanisms of cortactin(CTTN)on the invasion and migration of esophageal squamous cell carcinoma(ESCC)cells.Methods CTTN was overexpressed or knocked down in EC109 and TE1 cells by lentivirus infection.Then Western blotting was used to detect the expression of metastasis-related molecules including intercellular cell adhesion molecule-1(ICAM1),matrix metallo proteinase(MMP)2,MMP9,and vascular endothelial growth factor-C(VEGF-C).The abilities of invasion and migration were assessed by Transwell invasion assay and scratch test,respectively.Co-immunoprecipitation(Co-IP)was used to further validate the interaction between CTTN and target protein.Results CTTN was successfully overexpressed or knocked down in EC109 and TE1 cells,and stable infected cells were screened by puromycin.Compared with control groups,CTTN overexpression caused a clear increase in relative expressions of ICAM1,MMP2,MMP9,and VEGF-C(all P<0.001),and the capabilities of cell invasion and metastasis were significantly enhanced(the number of penetrating cells:541.0±17.8 vs.230.8±10.9,scratch healing rate:72.8%±1.9%vs.42.8%±1.6%,both P<0.001).CTTN knockdown decreased the relative expressions of ICAM1,MMP2,MMP9,and VEGF-C,and suppressed the capabilities of invasion and migration when compared with controls(the number of penetrating cells:102.3±4.0 vs.223.8±12.1,scratch healing rate:25.7%±1.5%vs.43.5%±1.9%,both P<0.001).The result of Co-IP showed that CTTN had obvious interaction with Src.Conclusion CTTN promotes the invasion and migration in ESCC cells through the interaction with Src.

关 键 词：皮动蛋白 受体蛋白质酪氨酸激酶类 SRC 食管肿瘤 癌 鳞状细胞 侵袭 转移 

分 类 号：R735.1[医药卫生—肿瘤]