摘要
目的探讨Gli-1在缺氧诱导乳腺癌MDA-MB-231细胞上皮-间质转分化(EMT)及侵袭中的重要作用。方法通过缺氧培养乳腺癌MDA-MB-231细胞,以常氧培养作为对照。Transwell小室侵袭试验检测各组MDAMB-231细胞的侵袭能力;Western blot检测HIF-1α、Gli-1、E-Cadherin和vimentin蛋白的表达水平。通过shRNA稳定转染乳腺癌细胞,再次通过Transwell小室侵袭试验检测缺氧对乳腺癌细胞侵袭能力的影响,Western blot检测HIF-1α、Gli-1、E-Cadherin和vimentin蛋白的表达水平。结果缺氧可明显诱导乳腺癌MDA-MB-231细胞侵袭,并上调HIF-1α、Gli-1和vimentin蛋白,下调E-cadherin蛋白。靶向沉默Gli-1基因后,缺氧失去了对乳腺癌细胞侵袭和EMT的诱导作用。结论缺氧通过上调Gli-1表达活化Hedgehog通路,诱导乳腺癌细胞侵袭及EMT过程。
Objective To investigate the important role of Gli-1 in hypoxia-induced epithelial-mesenchymal transition of breast cancer cell line MDA-MB-231. Methods For the present study, breast cancer cell line MDA- MB-231 was cultured under normoxic or hypoxic condition. Then, the shRNA of Gli-1 was stably transfected into MDA-MB-231 cells. Invasion of MDA-MB-231 was detected by the Transwell assay. The expressions of HIF-1α, Gli-1, E-Cadherin and vimentin proteins were determined by Western blot. Results Hypoxia significantly induced the invasion of MDA-MB-231 cells, upregulated the expressions of HIF-1α, Gli-1 and vimentin proteins, and downregulated the expression of E-Cadherin protein. Furthermore, knocking down Gli-1 obviously abrogated hypoxia-induced epithelial-mesenchymal transition and enhanced invasion of MDA-MB-231 cells. Gonclusion Hypoxia induces epithelial-mesenchymal transition and enhances invasion of breast carcinoma cells via strengthening Gli-1 expression and activation of Hedgehog pathway.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第6期800-804,819,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.81201824)
中央高校基本科研业务费专项资金资助项目(No.2013jdhz33)~~
