摘要
目的探讨Raptor在乳腺癌侵袭与转移过程中的作用。方法采用Western blot检测乳腺癌细胞MCF-7和MDA-MB-231(MDA231)中Raptor蛋白的表达情况。应用小干扰RNA技术下调乳腺癌细胞MDA231中Raptor的表达量,同时将过表达质粒转入乳腺癌细胞MCF-7中,应用Western blot法检测转染后各组细胞中上皮性钙黏着蛋白(E-cadherin)和波形蛋白(Vimentin)的表达;应用Transwell体外侵袭实验检测转染后各组细胞的侵袭能力;应用细胞核质分离实验检测转染后各组细胞中Snail蛋白的转核情况。结果MCF-7细胞中Raptor蛋白的表达量明显低于MDA231细胞中的表达量;转染后,与对照组Scr/MDA231相比,siRaptor/MDA231组中Raptor蛋白的表达量明显降低,Vimentin蛋白表达量降低,而E-cadherin蛋白表达量升高;与对照组MCF-7/Con相比,MCF-7/Raptor组中Raptor蛋白的表达量明显升高,并伴有Vimentin蛋白表达量升高,而Ecadherin蛋白表达量降低。siRaptor/MDA231细胞组中穿过Matrigel小室的细胞数量明显减少(P<0.05);MCF-7/Raptor细胞组中穿过Matrigel小室的细胞数量明显升高(P<0.05)。细胞核质分离实验结果显示,siRaptor/MDA231组中Snail蛋白表达量较Scr/MDA231组中明显降低;MCF-7/Raptor组中Snail蛋白表达量较MCF-7/Con组中明显升高。结论 Raptor能够促进乳腺癌EMT的发生,对乳腺癌的侵袭与转移有重要作用。
Aim To study the molecular mechanism of Raptor in the migration and invasion of breast cancer cells and provide the clinical theory basis for prevention of breast cancer invasion and metastasis. Methods Western blot was used to detect the expression of Raptor protein in MCF-7 cells and MDA231 cells. The siRNA plasmids were used to transfect MDA231 cells.At the same time,the plasmid pc DNA3. 1-Raptor was transfected into MCF-7 breast cancer cells. And Western blot was used to analyze the protein expression level of E-cadherin and Vimentin. Transwell was used to test the ability of invasion. Nucleus mass separation experiment was used to test the expression of Snail. Results The expression of Raptor protein in MDA231 cells was higher than in the MCF-7 cell. When the control group of Scr/MDA231 cells compared with siRaptor/MDA231, Raptor protein expression was decreased obviously after plasmid transfection interference,accompanied by reduction of Vimentin protein expression and increase of E-cadherin protein expression. Compared with MCF-7/Con,Raptor protein expression significantly increased in MCF-7/Raptor,accompanied by increase of Vimentin protein expression and reduction of E-cadherin protein expression. The number of cells through the artificial basilemma Transwell in siRaptor/MDA231 cells was significantly reduced（ P〈0. 05）,and the number of cells through the Transwell chambers artificial basilemma was significantly increased in MCF-7/Raptor（ P〈0. 05）. Nucleus mass separation experiment results showed that the expression of Snail decreased obviously in the siRaptor/MDA231 than in the Scr/MDA231,however,the expression of Snail was obviously higher in the MCF-7/Raptor. Conclusions Raptor can promote the occurrence of EMT in breast cancer,thus it promotes invasion and metastasis of breast cancer.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2017年第8期1091-1095,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金青年基金项目(No 81402389
81641111)
山东省自然科学基金资助项目(No ZR2014HL077
ZR 2015HL065)
山东省高等学校科技计划(No J12LK03
J13LK03)
关键词
RAPTOR
上皮-间质转化
乳腺癌
侵袭
转移
核质分离
Raptor
epithelial-mesenchymal transition
breast cancer
invasion
metastasis
nucleus mass separation
