下调Rab11基因对宫颈癌HeLa/SiHa细胞侵袭迁移的影响及机制探讨  

Effect of silencing Rab11 by RNAi on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism

作　　者：阚岩岩 张建华[1] 周敏[1] 章龙珍[2] 王侠[2] 

机构地区：[1]徐州医学院研究生学院肿瘤学系,江苏徐州221002 [2]徐州医学院附属医院放疗科,江苏徐州221002

出　　处：《中国癌症杂志》2016年第3期238-244,共7页China Oncology

基　　金：江苏省徐州市科技发展基金(KC14SH111)

摘　　要：背景与目的:Rab11基因在宫颈癌细胞中高表达,可能与细胞恶性转化相关。本研究采用RNA干扰技术下调Rab11基因表达,并探讨其对宫颈癌HeLa/SiHa细胞侵袭迁移的影响及可能的相关机制。方法:将HeLa/SiHa细胞分为2组:阴性对照组(转染阴性对照的小干扰RNA)和Rab11 siRNA干扰组(转染小干扰Rab11siRNA)。蛋白[质]印迹法(Western blot)检测Rab11干扰效果,检测转染Rab11 si RNA后,侵袭相关蛋白Rac1、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和MMP9表达的变化;细胞划痕损伤实验检测细胞迁移;Transwell实验检测细胞侵袭;细胞免疫荧光实验从形态学角度探索在小干扰Rab11后Rac1蛋白在细胞膜上聚集位置的变化。结果:转染小干扰Rab11siRNA到He La/SiHa细胞后,Rab11蛋白表达受到高效抑制(P<0.01);细胞迁移、侵袭能力受到抑制(P<0.05),Rac1蛋白表达明显下调(P<0.01),MMP2、MMP9蛋白表达下调(P<0.05),Rab11 siRNA处理组细胞运动前端细胞膜下Rac1蛋白聚集量减少。结论:Rab11基因表达下调可以抑制HeLa/SiHa细胞的迁移和侵袭,其机制可能与Rac1、MMP2和MMP9蛋白表达量下降及Rac1定位的改变有关。Background and purpose： The expression ofRabl 1 gene was increased in cervical cancer cell and may be involved in the cellular malignant transformation. This study used the sequence-specific siRNA knocking down the expression ofRabl 1 gene and aimed to investigate its effect on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism. Methods： HeLa/SiHa cells were divided into 2 groups： non-specific siRNA group transfected with unrelated siRNA （Rab 11-NC） and Rab 11 siRNA group transfected with Rab 11 siRNA （Rab 11 siRNA）. Western blot was used to examine the Rabl 1 protein expression. Cell migration and invasion were detected by cell scratch and Transwell invasion assay. Western blot was used to further investigate the expression of Racl, matrix metalloproteinase 2 （MMP2） and MMP9 which were critical for regulating cell invasion, Moreover, immunofluorescence was used to identify intracellular location of Racl in HeLaJSiHa cells. Results： The Rabl 1 siRNA inhibited expression of Rab 11 gene （P〈0.01）. The invasion and migration capacities of HeLa/SiHa cells were markedly inhibited in Rabl 1 siRNA group （P〈0.05）. The expression of Racl significantly decreased （P〈0.01）. The expression of MMP2 and MMP9 decreased （P〈0.05） as well. The recruitment of Rac 1 to protruding edge significantly decreased following down-regulation of Rab 11. Conclusion： Down-regulated Rab 11 expression could inhibit the expression of Rac 1, MMP2 and MMP9, and alter the location of Rac 1, leading to suppression of HeLa/SiHa cells migration and invasion.

关 键 词：Rab11 RAC1 基质金属蛋白酶 宫颈癌 细胞侵袭 细胞迁移 

分 类 号：R737.33[医药卫生—肿瘤]