【摘要】 目的 探讨miR-98在下咽癌Fadu细胞中的表达及其调控癌细胞转移及其上皮-间充质转化(EMT)的作用机制。方法 收集我院2016年10月～2021年1月临床确诊下咽癌患者的35例下咽癌组织及其28例癌旁正常组织，利用实时荧光定量PCR检测miR-98、EZH2其mRNA表达。通过转染技术下调miR-98表达，分别利用CCK-8法、Transwell和Western blot实验检测人下咽癌Fadu细胞的增殖、侵袭和上皮-间质质转化能力。通过生物信息学软件TargetScan分析miR-98的下游调控靶点，双荧光素酶报告基因实验进一步检测miR-98和EZH2之间的关系。通过转染技术下调EZH2表达，分析下调EZH2对Fadu细胞增殖、侵袭和EMT的影响。同时上调miR-98与EZH2表达，检测过表达miR-98通过EZH2对Fadu细胞转移及EMT的影响。下调或过表达EZH2后，Western blot检测Fadu细胞内JAK、p-JAK、STAT3和p-STAT3蛋白表达量。利用抑制剂AG490作用Fadu细胞，通过转染技术上调EZH2表达，检测Fadu细胞增殖、侵袭和EMT能力。结果 与癌旁正常组织相比，下咽癌组织中miR-98表达明显降低(P<0.01),EZH2表达明显上升(P<0.01)。下调miR-98促进了Fadu细胞的增殖、侵袭与EMT;miR-98靶向EZH2,和EZH2具有负调控关系；上调miR-98通过EZH2抑制了Fadu细胞的增殖、侵袭与EMT;下调EZH2抑制了Fadu细胞转移和EMT;上调EZH2激活了Fadu细胞内JAK/STAT3通路，且抑制剂AG490作用Fadu细胞能明显抑制过表达EZH2对Fadu细胞增殖、侵袭和EMT的上调作用。结论 miR-98靶向EZH2激活JAK/STAT3通路调控下咽癌细胞转移及其上皮-间充质转化。更多还原

【Abstract】 Objective To investigate the expression of miRNA-98 in hypopharyngeal carcinoma Fadu cells and the mechanism of its regulation on metastasis and epithelial mesenchymal transformation.Methods Real-time quantitative PCR was used to detect the mRNA expression of miR-98 and EZH2 in 35 hypopharyngeal carcinoma tissues and 28 adjacent normal tissues. The expression of miR-98 was down-regulated by transfection technique. The proliferation, invasion and epithelial mesenchymal transformation ability of Fadu cells were detected by CCK-8 assay, Transwell assay and Western blot assay, respectively. The downstream regulatory targets of miR-98 were analyzed by TargetScan, and the relationship between miR-98 and EZH2 was further detected by dual luciferase reporter assay. The expression of EZH2 was down-regulated by transfection technique, and the effects of down-regulation of EZH2 on proliferation, invasion and EMT of Fadu cells were analyzed. At the same time, the expression of miR-98 and EZH2 were up-regulated, and the effects of over-expression of miR-98 on Fadu cell metastasis and EMT through EZH2 were detected. After the down-regulation or overexpression of EZH2, the protein expressions of JAK, p-JAK, STAT3 and p-STAT3 in FADU cells were detected by Western blot. Fadu cells were treated with the inhibitor AG490, and the expression of EZH2 was up-regulated by transfection technology to detect the proliferation, invasion and EMT ability of Fadu cells.Results Compared with adjacent normal tissues, miR-98 expression was significantly reduced in hypopharyngeal carcinoma tissues(P<0.01), the expression of EZH2 was significantly increased(P<0.01).Downregulation of miR-98 promoted proliferation, invasion and EMT of Fadu cells. miR-98 targeted EZH2 and had a negative regulatory relationship with EZH2.The up-regulation of miR-98 inhibited the proliferation, invasion and EMT of Fadu cells by EZH2.Downregulation of EZH2 inhibited Fadu cell metastasis and EMT. The up-regulation of EZH2 activated the JAK/STAT3 pathway in Fadu cells, and AG490 can significantly inhibit the up-regulation of EZH2 overexpression on proliferation, invasion and EMT of Fadu cells. Conclusion miRNA-98 targeting EZH2 activates the JAK/STAT3 pathway to regulate hypopharyngeal cancer cell metastasis and epithelial-mesenchymal transformation.更多还原