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目的:为明确COX-2表达是否可以影响乳腺癌细胞上皮间质转化(EMT)变化,培养高表达的COX-2和EMT改变的乳腺癌MDA-MB-231细胞,用不同浓度的塞来昔布和小干扰RNA分别来抑制COX-2功能和表达,观测细胞EMT的变化和Snail的表达。方法:培养乳腺癌MDA-MB-231细胞,构建干扰COX-2 siRNA载体,将干扰载体转染MDAMB-231细胞,应用免疫荧光染色检测E-Cadherin、viminten和COX-2的表达,RT-PCR检测Western Blot检测Snail、COX-2的表达。检测培养液PGE2的水平。结果:小干扰RNA与塞来昔布治疗组相比E-Cadherin表达增加明显,viminten、Snail表达降低更显著,小干扰RNA处理乳腺癌MDA-MB-231细胞72 h后与塞来昔布处理的相比获得更多的上皮特征和更少的间质结构的特征。COX-2 mRNA在60μmol/L的塞来昔布处理组和RNAi处理组COX-2 mRNA的相对表达分别为0.97和0.22,塞来昔布不能抑制COX-2 mRNA表达。干扰组与塞来昔布组相比,细胞培养液中PGE2的浓度没有显著差异。结论:①COX-2的抑制可通过PGE2的下调调控乳腺癌EMT。②COX-2通过非PGE2依赖途径调节Snail的表达进而调控EMT的发生。
OBJECTIVE: To determine whether COX-2 expression can affect the epithelial-to-mesenchymal transition (EMT) in breast cancer cells and to culture the breast cancer MDA-MB-231 cells with high expression of COX-2 and EMT, with different concentrations of Celecoxib Cloned and small interfering RNA, respectively, to inhibit COX-2 function and expression were observed EMT changes and Snail expression. Methods: Breast cancer MDA-MB-231 cells were cultured and transfected into MDAMB-231 cells interfered with COX-2 siRNA vector. The expression of E-Cadherin, viminten and COX-2 was detected by immunofluorescence staining. Western Blot was used to detect the expression of Snail and COX-2. The level of PGE2 in culture medium was detected. Results: Compared with Celecoxib group, the expression of E-Cadherin increased significantly and the expression of viminten and Snail decreased more significantly. Small interfering RNA treatment of breast cancer MDA-MB-231 cells 72 hours after treatment with celecoxib More epithelial features and fewer features of interstitial structures. The relative expression of COX-2 mRNA of COX-2 mRNA in 60μmol / L Celecoxib treated group and RNAi-treated group was 0.97 and 0.22, respectively. Celecoxib could not inhibit the expression of COX-2 mRNA. There was no significant difference in the concentration of PGE2 in the cell culture medium between the interferon group and the celecoxib group. Conclusion: (1) The inhibition of COX-2 can regulate breast cancer EMT through the down-regulation of PGE2. ②COX-2 regulates the occurrence of EMT by regulating the expression of Snail through non-PGE2-dependent pathway.