姜黄素抑制耐紫杉醇食管癌细胞的上皮间质转化作用及机制探讨

发布时间：2018-05-24 03:10

本文选题：紫杉醇 + 食管肿瘤　；参考：《重庆医学》2017年34期

【摘要】：目的建立耐紫杉醇食管癌(EC)细胞系EC9706/PTX,观察姜黄素对EC9706/PTX细胞上皮间质转化(EMT)的抑制作用并探讨其机制,为耐药EC的治疗提供理论依据。方法用中等浓度间歇作用法建立耐紫杉醇EC细胞EC9706/PTX,四甲基偶氮唑蓝(MTT)法测定细胞耐药指数及交叉耐药性,检测不同浓度姜黄素对EC9706/PTX细胞增殖的抑制。使用细胞骨架染色、划痕实验、Transwell侵袭实验检测姜黄素对EC9706/PTX细胞形态变化、迁移和侵袭能力的影响。荧光定量PCR及蛋白免疫印迹(Western blot)检测姜黄素对EC9706/PTX细胞中EMT相关分子标志物E-钙黏蛋白、N-钙黏蛋白、波形蛋白、纤维连接蛋白在mRNA和蛋白水平的表达影响。Western blot检测姜黄素对EC9706/PTX细胞中转录因子NF-κB p65及Snail在蛋白水平的表达影响。结果 EC9706/PTX对紫杉醇的耐药指数为28.4,对顺铂、阿霉素产生交叉耐药性,姜黄素能够抑制EC9706/PTX细胞的增殖。在20μmol/L浓度的姜黄素作用下,EC9706/PTX细胞的迁移和侵袭能力明显降低。荧光定量PCR及Western blot检测显示,细胞耐药后E-钙黏蛋白的表达明显下调,而N-钙黏蛋白表达则明显上调,姜黄素逆转了这一现象。Western blot检测提示,EC细胞发生耐药及EMT后,NF-κB p65及Snail蛋白的表达增强,姜黄素阻断了这一作用。结论姜黄素能够抑制紫杉醇耐药EC细胞的增殖并且能够逆转紫杉醇耐药EC细胞的EMT现象,其机制可能是通过抑制NF-κB-Snail信号通路实现的。
[Abstract]:Objective to establish paclitaxel-resistant esophageal carcinoma cell line EC9706 / PTX. to observe the inhibitory effect of curcumin on epithelial mesenchymal transformation of EC9706/PTX cells and its mechanism, and to provide a theoretical basis for the treatment of drug-resistant EC. Methods Taxol-resistant EC cells EC9706 / PTX and tetramethylazolam MTT were used to determine the drug resistance index and cross resistance. The inhibition of curcumin at different concentrations on the proliferation of EC9706/PTX cells was detected. The effects of curcumin on the morphological changes, migration and invasion of EC9706/PTX cells were detected by cytoskeleton staining and transwell invasion assay. Fluorescence quantitative PCR and Western blot were used to detect the effect of curcumin on EMT related molecular marker E-cadherin and vimentin in EC9706/PTX cells. The expression of fibronectin at mRNA and protein levels. Western blot was used to detect the expression of transcription factor NF- 魏 B p65 and Snail in EC9706/PTX cells. Results the resistance index of EC9706/PTX to paclitaxel was 28. 4, cross resistance to cisplatin and adriamycin. Curcumin could inhibit the proliferation of EC9706/PTX cells. The migration and invasion ability of EC9706 / PTX cells was significantly decreased at 20 渭 mol/L concentration of curcumin. Fluorescence quantitative PCR and Western blot analysis showed that the expression of E-cadherin was down-regulated, while the expression of N- cadherin was up-regulated. Curcumin reversed this phenomenon. Western blot assay showed that the cells developed drug resistance and the expression of NF- 魏 B p65 and Snail protein increased after EMT, which was blocked by curcumin. Conclusion curcumin can inhibit the proliferation of paclitaxel-resistant EC cells and reverse the EMT phenomenon of paclitaxel-resistant EC cells. The mechanism may be through the inhibition of NF- 魏 B-Snail signaling pathway.
【作者单位】：郑州大学附属肿瘤医院/河南省肿瘤医院消化内一科;
【基金】：河南省科技厅基础与前沿项目(112300410044)
【分类号】：R735.1

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