解剖学报

解剖学报 ›› 2017, Vol. 48 ›› Issue (6): 682-687.doi: 10.16098/j.issn.0529-1356.2017.06.008

• 肿瘤生物学 • 上一篇    下一篇

Raptor通过Wnt3a/β-catenin信号通路对乳腺癌侵袭与转移的作用

 徐新伟1 杨玉玲1 孙志亮2 张国新2 曾国栋2 李洪利3 尹崇高4*   

  1. 1. 潍坊医学院病理学教研室; 2. 生物科学与技术学院; 3. 医学研究实验中心; 4. 护理学院，山东 潍坊 261053
  • 收稿日期:2017-03-06 修回日期:2017-05-02 出版日期:2017-12-06 发布日期:2017-12-06
  • 通讯作者: 尹崇高 E-mail:lihongli@ufmc.edu.cn
  • 基金资助:
    国家自然科学基金青年基金项目;山东省自然科学基金;山东省高等学校科技计划

Raptor promoting invasion and metastasis of breast cancer through the signaling pathway Wnt3a/β-catenin

XU Xin-wei1 YANG Yu-ling1 SUN Zhi-liang2 ZHANG Guo-xin2 ZENG Guo-dong2 LI Hong-li3 YIN Chong-gao 4*   

  1. 1.Department of Pathology;2.College of Biological Science and Technology; 3. Medicine Research Center; 4. Colloge of Nursing, Weifang Medical University, Shandong Weifang 261053, China
  • Received:2017-03-06 Revised:2017-05-02 Online:2017-12-06 Published:2017-12-06
  • Contact: YIN Chong-gao E-mail:lihongli@ufmc.edu.cn

PDF

摘要：

目的 探讨Raptor是否可以通过Wnt3a/β-catenin信号通路促进乳腺癌细胞的侵袭与转移。 方法 采用免疫印迹法（Western blotting）检测乳腺癌细胞MCF-7和MDA-MB-231(MDA231)中Raptor蛋白的表达情况；采用小RNA（RNAi）干扰技术将质粒转染MDA231，将过表达质粒转染MCF-7；趋化运动实验检测乳腺癌细胞的运动能力；Transwell侵袭实验检测乳腺癌细胞的侵袭能力；Western blotting检测上皮细胞间充质转化（EMT）标志物的表达情况；核质分离实验检测β-联蛋白（β-catenin）的转核情况。 结果 Western blotting法检测结果显示，Raptor在低转移细胞系MCF-7中低表达，而在高转移细胞系MDA231中高表达。转染后，SiRaptor/MDA231细胞组中Raptor蛋白的表达明显下降，MCF-7/Raptor细胞组中Raptor蛋白的表达明显升高，表明转染成功。用Wnt3a刺激处理后，趋化运动和Transwell侵袭实验结果显示，SiRaptor/MDA231细胞组运动和侵袭能力明显降低，MCF-7/Raptor细胞组运动和侵袭能力明显增强。Western blotting法检测结果显示，用Wnt3a和抑制剂Dkk1不同处理后，SiRaptor/MDA231细胞组中上皮-钙黏蛋白（E-cadherin）上调，波形蛋白（vimentin）下调，细胞核中β-0catenin的表达降低，而MCF-7/Raptor细胞组中E-cadherin蛋白下调，vimentin蛋白上调，β-catenin蛋白的表达升高。 结论 Raptor能通过Wnt3a/β-catenin信号通路促进EMT的发生，进而促进乳腺癌细胞的侵袭与转移。

关键词: 乳腺癌, Raptor, Wnt3a/β-catenin, 上皮细胞-间充质转化, 侵袭, 免疫印迹法

Abstract:

Objective To explore whether the Raptor can promote invasion and metastasis of breast cancer cells through Wnt3a/-catenin signaling pathway. Methods Using Western blotting to detect the protein expression of Raptor in breast cancer cells MDA-MB-231(MDA231) and MCF-7; Using RNAi technology to transfect small RNA interference plasmid into MDA231 breast cancer cells transiently. Overexpression of plasmid was transfected into MCF-7 breast cancer cells. Transwell and chemotaxis assay were used to detect the invasion and chemotaxis ability of breast cancer cells. Western blotting detected the expression of epithelial-mesenchymal transition(EMT) markers; Karyoplasm separation experiment tested the expression of β-catenin in nuclei. Results Western blotting result showed that the expression of Raptor was low in MCF-7, and high in MDA231. The protein expression of Raptor in SiRaptor/MDA231 was decreased obviously, and the protein expression of Raptor in MCF-7/Raptor was obviously higher. Transwell invasion and chemotaxis assay result showed that the invasion and chemotaxis ability of SiRaptor/MDA231 was decreased with the stimulation of Wnt3a, however, its capability of invasion and chemotaxis of MCF-7/Raptor was significantly enhanced. According to the result of Western blotting method with unused Wnt3a or at the same time stimulated with Wnt3a and Dkk1, the expression of E-cadherin was higher, and the expressions of vimentin and β-catenin were reduced in SiRaptor/MDA231, while in the MCF-7/Raptor, the expression of E-cadherin was reduced, and the expressions of vimentin and β-catenin were increased. Conclusion Through Wnt3a/β-catenin signaling pathway Raptor promotes the occurrence of EMT, thus to promote the invasion and metastasis of breast cancer cells.

Key words: Breast cancer, Raptor, Wnt3a/β-catenin, Epithelial-mesenchymal transition, Invasion, Western blotting