摘要
目的研究Hedgehog信号通路对乳腺癌细胞侵袭能力的影响及其可能机制。方法通过Western blot和Real-time RT-PCR,检测人乳腺癌细胞MDA-231和人正常乳腺上皮细胞MCF-10A中SHH、SMO和Gli-1蛋白和mRNA表达。通过小干扰RNA转染MDA-231,以未转染细胞为正常对照组,转染siRNA Control为阴性对照组,Western blot和RT-PCR法检测靶向沉默SMO基因的效果,Transwell小室侵袭试验检测各组MDA-231细胞的侵袭能力,Western blot检测Gli-1、Snail、MMP-9、E-cadherin和Vimentin的表达变化。结果人乳腺癌细胞MDA-231高表达SHH、SMO和Gli-1。通过RNA干扰技术靶向沉默MDA-231细胞SMO基因,其细胞侵袭能力显著下调,其Gli-1、Snail、MMP-9、Vimentin表达下调,E-cadherin表达增强。结论 Hedgehog通路的异常激活与乳腺癌细胞侵袭相关,其机制可能与诱导乳腺癌上皮细胞间质转分化有关。
Objective To investigate the effect and mechanism of Hedgehog signaling pathway on the invasion of breast cancer cells in vitro. Methods The SHH, SMO and Gli-1 expression levels of breast cancer cell line MDA-231 and normal mammary epithelial cell line MCF-10A were detected by Western blot at protein level and by Real-time RT-PCR at mRNA level. Next, shRNA vector was transfected into the MDA-231 cells with highly expressed SMO, and the stable transfected cells were selected by G418. Western blot and Real-time RT-PCR were performed to observe the inhibitory effect of RNAi on SMO expression. MTT assay was used to assess the influence of SMO siRNA on cell proliferation. Transwell assay was applied to observe cell invasion ability. The expressions of Gli-1, Snail, MMP-9, E-cadherin and Vimentin protein were determined by Western blot. Restlits Compared with those of normal mammary epithelial cell line MCF-10A, the expressions of SHH, SMO and Gli-1 were significantly increased. The invasion of MDA-231 cells was inhibited significantly after SMO silencing. Additionally, the protein expressions of Gli-1, Snail, MMP-9 and Vimentin were obviously inhibited, and E-cadherin was significantly increased. Conclusion Mutative activation of Hedgehog signaling pathway in breast cancer cells promotes cell invasion probably through induction of epithelial-mesenchymal transition of the tumor cells.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2017年第1期48-52,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.81302029
81572734)
陕西省自然科学基金资助项目(No.2014JQ4149)
陕西省社会发展科技攻关项目(No.2016SF-121)~~
