摘要
目的探讨β-连环蛋白(β-catenin)对乳腺癌细胞系MDA-MB-231细胞上皮间质表型和侵袭迁移能力的影响。方法用β-catenin si RNA质粒转染乳腺癌MDA-MB-231细胞(对照组),免疫荧光染色观察β-catenin的表达情况。Western blot检测上皮标记蛋白E-cadherin、间质标记蛋白vimentin及上皮间质转化相关调控因子Twist1和Snail的表达。Transwell侵袭实验和伤口愈合实验比较对照组、空质粒组和β-catenin下调组细胞体外侵袭迁移能力。结果免疫荧光染色显示对照组和空质粒组细胞可见β-catenin在胞膜、胞浆和胞核表达,β-catenin下调组细胞胞膜β-catenin呈低表达,且未见胞浆、胞核表达。下调β-catenin的细胞E-cadherin表达增高,而vimentin、Twist1和Snail表达减少。侵袭迁移实验证明,下调组细胞穿膜细胞数和迁移距离显著少于MDA-MB-231和对照组(P<0.05)。结论下调β-catenin阻碍Wnt/β-catenin通路的激活,降低乳腺癌MDA-MB-231细胞间质表型的表达而促进其上皮表型的表达,从而降低了细胞的体外侵袭、迁移能力。
Objective To investigate the effect ofβ-catenin on epithelial mesenchymal phenotype and invasion migration ability of breast cancer cell line MDA-MB-231. Methods Small interfering RNA （siRNA） targetingβ-catenin plasmid was transfected into MDA-MB-231 cell line. Immunofluorescence staining was used to observe the expression of β-catenin. The expressions of epithelial cell marker E-cadherin and mesenchymal marker vimentin, epithelial mesenchymal transition correlation factor Twist1 and Snail were detected by Western blot assay. Invasion and migration ability was compared by transwell invasion and wound healing assay between control group, the MDA-MB-231 group andβ-catenin down-regulated group. Results Immunofluorescence staining showed thatβ-catenin was expressed in cell membrane, cytoplasm or nucleus in MDA-MB-231 group and control group. There was a decreased expression in β-catenin down-regulated group, and no expression in cytoplasm or nucleus. The expression of E-cadherin was increased, while vimentin, Twist 1 and Snail expression decreased inβ-catenin down-regulated cells. Transwell invasion and wound healing assay results proved that transmembrane cell number and migration distance were significantly lower in β-catenin down-regulated group than those of MDA-MB-231 group and control group （P〈0.05）. Conclusion The down-regulation ofβ-catenin inhibits Wnt/β-catenin activation that decreases the mesenchymal phenotype but increases epithelial phenotype of breast cancer cells MDA-MB-231, and which reduces the cell invasion and migration ability in vitro.
出处
《天津医药》
CAS
2015年第9期981-984,I0005,共5页
Tianjin Medical Journal
