miR-98-5p靶向CCR7对乳腺癌细胞MCF-7运动能力的影响及其机制研究

Effect of miR-98-5p Targeting CCR7 on the Mobility of Breast Cancer Cell MCF-7 and Its Related Mechanism

关键词:  乳腺癌  miR-98-5p  趋化因子受体  上皮间充质转化

Key words:  breast cancer  miR-98-5p  CC-chemokine receptor 7  epithelial-mesenchymal transition

摘要:

目的 探究miR-98-5p靶向作用于趋化因子受体（CC chemokine receptor 7，CCR7）对乳腺癌细胞MCF-7运动能力的影响及相关机制。方法 MCF-7细胞分为对照组、miR-98-5p mimic组、miR-98-5p NC组、pc-CCR7组、miR-98-5p+pc-CCR7组，分别转染相应的miRNA和CCR7过表达载体，RT-PCR检测miR-98-5p和CCR7基因表达水平，荧光素酶报告试验检测miR-98-5p和CCR7靶向关系，Transwell法检测细胞侵袭，划痕法检测细胞迁移，Western blotting检测CCR7、基质金属蛋白酶2（MMP-2）、MMP-9、血管内皮生长因子（VEGF）、E-钙黏着蛋白（E-cadherin）、N-cadherin、波形蛋白（Vimentin）蛋白表达水平。结果 与CCR7 WT+miR-98-5p NC比较，CCR7 WT+miR-98-5p mimic荧光素酶活性显著降低。与对照组比较，miR-98-5p mimic组中miR-98-5p表达水平、E-cadherin蛋白表达水平显著升高，CCR7基因和蛋白表达水平、细胞侵袭和迁移能力、MMP-2、MMP-9、VEGF、N-cadherin、Vimentin蛋白表达水平显著降低，pc-CCR7组细胞侵袭和迁移能力、CCR7、MMP-2、MMP-9、VEGF、N-cadherin、Vimentin蛋白表达水平显著升高，E-cadherin蛋白表达水平显著降低；与pc-CCR7组比较，miR-98-5p+pc-CCR7组细胞侵袭和迁移能力、CCR7、MMP-2、MMP-9、VEGF、N-cadherin、Vimentin蛋白表达水平显著降低，E-cadherin蛋白表达水平显著升高。结论 miR-98-5p可靶向作用于CCR7，抑制乳腺癌细胞MCF-7运动能力，其作用机制与上皮细胞间充质转化有关。

Abstract:

OBJECTIVE To investigate the effect and mechanism of miR-98-5p on the mobility of breast cancer cell MCF-7 by targeting CC chemokine receptor 7(CCR7). METHODS MCF-7 cells were divided into control, miR-98-5p mimic, miR-98-5p NC, pc-CCR7, miR-98-5p+pc-CCR7 groups, each group was treated with corresponding miRNA and CCR7 overexpression vector, gene levels of miR-98-5p and CCR7 were measured by RT-PCR, luciferase reporter assay was performed for measuring the relationship between miR-98-5p and CCR7, invasion ability was observed by Transwell, migration was observed by cell scratch. Western blotting was used to determine the protein levels of CCR7, MMP-2, MMP-9, VEGF, E-cadherin, N-cadherin, Vimentin. RESULTS In the Luciferase reporter assay, the luciferase activity of CCR7 WT+miR-98-5p mimic group was significantly reduced compared to CCR7 WT+miR-98-5p NC group. Compared with control group, the level of miR-98-5p, protein level of E-cadherin were increased notably, and the gene and protein levels of CCR7, cell invasion and migration ability, protein levels of MMP-2, MMP-9, VEGF, N-cadherin, Vimentin were decreased significantly in miR-98-5p mimic group, the cell invasion and migration ability, protein levels of CCR7, MMP-2, MMP-9, VEGF, N-cadherin, Vimentin were increased markedly, and the protein level of E-cadherin decreased notably in pc-CCR7 group. Compared with pc-CCR7 group, the cell invasion and migration ability, protein levels of CCR7, MMP-2, MMP-9, VEGF, N-cadherin, Vimentin were decreased markedly, and the protein level of E-cadherin was increased significantly. CONCLUSION miR-98-5p can target CCR7, to inhibit the mobility of breast cancer cell MCF-7, and the mechanism related to epithelial-mesenchymal transition.