摘要
目的通过TGF-β1诱导乳腺癌MCF-7发生上皮-间质转化(epithelial-mesenchymal transition,EMT)后检测锌指转录因子Snail表达的改变,探讨Snail在EMT及乳腺癌发生发展中的作用。方法常规培养乳腺癌细胞株MCF-7后,用TGF-β1诱导其发生EMT,用Transwell侵袭小室法进行细胞体外侵袭能力检测;用免疫组织化学方法及免疫荧光检测E-cadherin、Vi mentin、Snail的表达;用real ti me PCR检测E-cadherin、Vi mentin、Snail mRNA的表达。结果TGF-β1处理72h后的MCF-7细胞穿透能力明显增强。E-cadherin蛋白及mRNA表达减少,Vi mentin、Snail蛋白及mRNA表达增加。结论E-cadherin、Vi mentin是细胞发生EMT的重要生物学标志,Snail可能在转录水平上调控E-cadherin、Vi mentin蛋白的表达,Snail在EMT和乳腺癌的发生发展中起着重要的作用。
Objective To detect the expression of the zinc finger transcription factor snail after TGF-β1-induced epithelial-mesenchymal transition（EMT）,and investigate the role of snail in EMT and the development of breast cancer.Methods TGF-β1 induced EMT in routinely cultured breast cancer cell line MCF-7.The invasive and metastatic capacity was evaluated with transwell migration assay.The expressions of E-cadherin,vimentin and snail protein and mRNA were investigated by real time PCR,immunohistochemistry and immunofluorescence.Results TGF-β1 significantly decreased the expressions of E-cadherin protein and mRNA,but increased the expressions of Vimentin and snail protein and mRNA.The cellular invasion assay indicated that TGF-β1 drastically enhanced the invasive potential of MCF-7.Conclusion E-cadherin and Vimentin are important biological markers of EMT.Snail may regulate the expressions of E-cadherin and Vimentin protein at the transcriptional level.Snail plays an important role in EMT and the development of breast cancer.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2011年第2期120-123,共4页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金(81072068)
山东省优秀中青年科学家科研奖励基金(BS2010yy071)
山东省潍坊医学院青年教师启动基金(KQ07037)
