中国癌症杂志 ›› 2016, Vol. 26 ›› Issue (3): 238-244.doi: 10.3969/j.issn.1007-3969.2016.03.006

• 论著 • 上一篇    下一篇

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

阚岩岩1,张建华1,周 敏1,章龙珍2,王 侠2   

  1. 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
  • 出版日期:2016-03-30 发布日期:2016-06-13
  • 通信作者: 王 侠 E-mail: wangxia_66@163.com
  • 基金资助:
    江苏省徐州市科技发展基金(KC14SH111)。

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

KAN Yanyan1, ZHANG Jianhua1, ZHOU Min1, ZHANG Longzhen2, WANG Xia2   

  1. 1. Department of Oncology, the Graduate School, Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China; 2.Department of Radiotherapy, the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China
  • Online:2016-03-30 Published:2016-06-13
  • Contact: WANG Xia E-mail: wangxia_66@163.com

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 siRNA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HeLa/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。

关键词: Rab11, Rac1, 基质金属蛋白酶, 宫颈癌, 细胞侵袭, 细胞迁移

Abstract: Background and purpose: The expression of Rab11 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 of Rab11 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 (Rab11-NC) and Rab11 siRNA group transfected with Rab11 siRNA (Rab11siRNA). Western blot was used to examine the Rab11 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 Rac1, matrix metalloproteinase 2 (MMP2) and MMP9 which were critical for regulating cell invasion. Moreover, immunofluorescence was used to identify intracellular location of Rac1 in HeLa/SiHa cells. Results: The Rab11 siRNA inhibited expression of Rab11 gene (P<0.01). The invasion and migration capacities of HeLa/SiHa cells were markedly inhibited in Rab11siRNA group (P<0.05). The expression of Rac1 significantly decreased (P<0.01). The expression of MMP2 and MMP9 decreased (P<0.05) as well. The recruitment of Rac1 to protruding edge significantly decreased following down-regulation of Rab11. Conclusion: Down-regulated Rab11 expression could inhibit the expression of Rac1, MMP2 and MMP9, and alter the location of Rac1, leading to suppression of HeLa/SiHa cells migration and invasion.

Key words: Rab11, Rac1, Matrix metalloproteinase, Cervical cancer, Cell invasion, Cell migration