Sa Na, Yu Liang, Mu Yakui, Sun Gaoying, Xu Wei
Department of Otorhinolaryngology Head and Neck Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China.
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Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2014 Dec;49(12):990-4.
To investigate the expression of microRNA-214(miR-214) in advanced hypopharyngeal carcinoma tissues and its effects on the invasion, migration and colone formation of FaDu cells.
miR-214 expression in 30 cases of advanced hypopharyngeal carcinoma tissues and normal hypopharyngeal mucosa tissues was detected by real-time quantitative polymerase chain reaction (RT-qPCR). miR-214 was upregulated through transfecting the overexpression vector hsa-mir-214 into FaDu cells. The influences of miR-214 upregulation on the invasion, migration, clone formation and Twist expression were measured by Transwell invasion, Transwell migration, plate clone formation and Western blot assays, respectively.
The expression of miR-214 in advanced hypopharyngeal carcinoma tissues (0.311 ± 0.206) was significantly less than normal hypopharyngeal mucosa tissues (1.620 ± 1.394; t = 5.09, P < 0.05) . The expression of miR-214 was notably upregulated after tranfected with hsa-mir-214 compared with the negative control group (t = 6.347, P < 0.05). The migration and invasion ability of FaDu cells transfeced with hsa-mir-214 was decreased by comparison with negative control cells (t = 11.6, P < 0.01; t = 6.499, P < 0.05). There was no significant difference of the average clony number and the cloning efficiency between the experimental and negative control groups (t = 0.592, P > 0.05).
of Western blot assay showed that, Twist expression in the miR-214-overexpressed group was apparently decreased compared with that in the control group (t = 6.545, P < 0.05).
miR-214 is expressed at a low level in advanced hypopharyngeal carcinoma tissues, and can obviously inhibit the invasion and migration abilities of FaDu cells, possibly because of its inhibiting effect on Twist expression. Additionally, miR-214 plays no significant role in the proliferation of FaDu cells.
探讨微小RNA-214（miR-214）在晚期下咽癌组织中的表达及其对FaDu细胞侵袭、迁移和克隆形成的影响。
采用实时定量聚合酶链反应（RT-qPCR）检测30例晚期下咽癌组织及正常下咽黏膜组织中miR-214的表达。通过将过表达载体hsa-mir-214转染至FaDu细胞来上调miR-214。分别采用Transwell侵袭实验、Transwell迁移实验、平板克隆形成实验和蛋白质免疫印迹法检测miR-214上调对细胞侵袭、迁移、克隆形成及Twist表达的影响。
晚期下咽癌组织中miR-214的表达量为（0.311±0.206），显著低于正常下咽黏膜组织（1.620±1.394；t=5.09，P<0.05）。与阴性对照组相比，转染hsa-mir-214后miR-214的表达显著上调（t=6.347，P<0.05）。与阴性对照细胞相比，转染hsa-mir-214的FaDu细胞迁移和侵袭能力降低（t=11.6，P<0.01；t=6.499，P<0.05）。实验组与阴性对照组之间的平均克隆数和克隆效率差异无统计学意义（t=0.592，P>0.05）。蛋白质免疫印迹法结果显示，miR-214过表达组中Twist的表达明显低于对照组（t=6.545，P<0.05）。
miR-214在晚期下咽癌组织中低表达，可明显抑制FaDu细胞的侵袭和迁移能力，可能是通过抑制Twist表达实现的。此外，miR-214对FaDu细胞的增殖无显著作用。
