Gong X R, Yu F, Zhou Y B
Institute of Otolaryngology Head and Neck Surgery Affiliated to Guangzhou Medical University, Guangzhou Hospital of Otolaryngology Head and Neck Surgery, Guangzhou 12th People's Hospital, Guangzhou, 510620, China.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2017 Dec 5;31(23):1839-1843. doi: 10.13201/j.issn.1001-1781.2017.23.014.
To investigate the mechanism between epithelial-mesenchymal transition (EMT) and cisplatin induced resistant cell subline and the malignant biological characteristics, to explore EMT in human hep-2 laryngeal resistant cells. Using cisplatin-resistant cells (hep-2/CDDP) and non-resistant cells (hep-2) established in our previous study; the invasion and migration biological behaviors were detected by transwell and scratch assay; the expressions of E-cadherin, Zo-1, Snail, Slug, Twist1, Vimentinon in the mRNA level were detected by RT-qPCR and the protein level by Western blot. Transwell and scratch assay show the invasion and migration behaviors were increased in hep-2/CDDP cells (<0.05), the epithelial marker E-cadherin and Zo-1 were downregulated in hep-2/CDDP cells (all <0.05), transcription factor Snail, Slug were upregulated in mRNA and protein level (all <0.01) while Twist1 had no significant changed in protein level (>0.05), the expression of mesenchymal marker Vimentin was also increased in mRNA and protein levels in cisplatin resistant cells (<0.01). It was confirmed that the hep-2/CDDP cells possessed EMT phenotypes. The cisplatin resistant laryngeal cancer cells perform higherinvasion and migration biological behaviors,and the mechanisms of increased ability of invasion and migration induced by cisplatin was associated to eEMT, study on signal path related to EMT may overcome cisplatin resistance and reduce invasion and migration behaviors.
探讨上皮-间质转化（EMT）与顺铂诱导的耐药细胞亚系及恶性生物学特性之间的机制，以研究人喉癌Hep-2耐药细胞中的EMT。采用本课题组前期建立的顺铂耐药细胞（Hep-2/CDDP）和非耐药细胞（Hep-2）；通过Transwell和划痕实验检测侵袭和迁移生物学行为；采用RT-qPCR检测E-钙黏蛋白、紧密连接蛋白1（Zo-1）、蜗牛蛋白（Snail）、蛞蝓蛋白（Slug）、Twist1、波形蛋白（Vimentin）在mRNA水平的表达，采用蛋白质免疫印迹法检测其在蛋白水平的表达。Transwell和划痕实验显示，Hep-2/CDDP细胞的侵袭和迁移能力增强（P<0.05），上皮标志物E-钙黏蛋白和Zo-1表达下调（均P<0.05），转录因子Snail、Slug在mRNA和蛋白水平均上调（均P<0.01），而Twist1蛋白水平无明显变化（P>0.05），顺铂耐药细胞中，间质标志物波形蛋白在mRNA和蛋白水平表达也增加（P<0.01）。证实Hep-2/CDDP细胞具有EMT表型。顺铂耐药喉癌细胞具有更高的侵袭和迁移生物学行为，顺铂诱导侵袭和迁移能力增强的机制与EMT相关，研究与EMT相关的信号通路可能克服顺铂耐药并降低侵袭和迁移行为。
