顺铂诱导上皮间质转化在耐药喉癌细胞中的作用

发布时间：2018-05-27 11:39

本文选题：上皮间质转化 + 顺铂　；参考：《临床耳鼻咽喉头颈外科杂志》2017年23期

【摘要】：目的:观察顺铂(CDDP)诱导的人喉癌耐药细胞(hep-2/CDDP)中的上皮间质转化(EMT)情况,探讨EMT在喉癌顺铂耐药中的机制。方法:利用hep-2细胞和前期培养的hep-2/CDDP细胞株;Transwell、划痕实验检测细胞的侵袭迁移能力;分别提取人喉癌顺铂耐药细胞hep-2/CDDP及其亲代细胞hep-2mRNA、总蛋白,应用实时荧光定量PCR(RT-qPCR)、Western blot方法检测上皮表型E-cadherin、Zo-1,上皮间质转化转录因子Snail、Slug、Twist1,间质表型分子Vimentin mRNA及蛋白表达水平。结果:Transwell结果显示耐药细胞穿过基质胶的数量高于亲本细胞(P0.05),划痕实验示hep-2/CDDP细胞划痕愈合面积百分比高于hep-2细胞(P0.05),hep-2/CDDP细胞上皮表型E-cad、Zo-1mRNA及蛋白表达水平较hep-2细胞降低(均P0.01),EMT转录因子Snail、Slug表达水平较hep-2细胞升高(均P0.01),Twist1蛋白水平表达无明显变化(P0.05),间质表型Vimentin mRNA及蛋白表达水平较hep-2细胞升高(均P0.01)。结论:hep-2细胞的耐药性和侵袭迁移能力改变与顺铂诱导的上皮间质转变可能有关,抑制EMT相关通路可能为逆转喉癌顺铂耐药提供新思路。
[Abstract]:Aim: to investigate the mechanism of EMT in cisplatin induced resistance to cisplatin in human laryngeal cancer cell line hep-2 / CDDP. Methods: the invasive and migration ability of hep-2 cells and hep-2/CDDP cell lines were tested by scratch test, and the total proteins of hep-2/CDDP cells and their parental cells, hep-2mRNAs, were extracted, respectively. The expression of E-cadherinine Zo-1, the transcriptional factor Snail-SlugSlugt1, the Vimentin mRNA and protein expression of the mesenchymal phenotypic molecules were detected by real-time fluorescence quantitative PCR- RT-qPCR- blot. The expression of E-cadherinine Zo-1, the transcriptional factor Slug1, and the expression of Vimentin mRNA and protein were detected. Results the results showed that the number of drug resistant cells passing through matrix glue was higher than that of parent cells P0.05.The scratch test showed that the percentage of scratch healing area of hep-2/CDDP cells was higher than that of hep-2 cells. The expression level of E-cadhe Zo-1 mRNA and protein in the epithelial phenotype of hep-2 cells was lower than that of hep-2 cells. The expression of Slug, a transcription factor of P0.01EMT, was higher than that of hep-2 cells. The expression of Twist1 protein was not changed significantly. The expression of Vimentin mRNA and protein in stromal cells was higher than that in hep-2 cells (all P0.01). Conclusion the changes of drug resistance and invasion and migration in the cell line may be related to the epithelial interstitial transition induced by cisplatin. Inhibition of EMT related pathway may provide a new way to reverse cisplatin resistance in laryngeal carcinoma.
【作者单位】：广州医科大学耳鼻咽喉头颈外科研究所广州市耳鼻咽喉头颈外科医院广州市第十二人民医院;
【基金】：广州市喉癌临床医学与转化中心试点建设项目(No:201604020005)
【分类号】：R739.65

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