MiR-124对鼻咽癌细胞株上皮间充质转化和放疗敏感性的影响

摘要

Objective To identify the role of miR-124 in regulating the radiosensitivity and the epithelial-mesenchymal transition (EMT) of nasopharyngeal carcinoma (NPC). Methods Transient transfection of cells with miR-124 mimic or inhibitor was performed and wound-healing assay was used to investigate the role of miR-124 in the EMT of NPC. The apoptosis affected by miR-124 was also measured after irradiation , followed by investigating the cell proliferation by EdU assay. Finally , proteins of Akt and ERK associated with EMT and radiosensitivity, were measured by western blot. Results The migration index from NPC cell line indicated that miR-124 repressed the EMT. The results from caspase-3 activity assay showed that caspase-3 activity after irradiation significantly increased in miR-124 mimic group compared with the control group (P < 0.01). It was also confirmed that irradiation led to a higher percentage of apoptosis in miR-124 group compared with the control group in NPC cells. Cell proliferation after irradiation was significantly decreased in MiR-124 group as compared with control group . MiR-124 inhibited the protein expression of p-Akt . Conclusion MiR-124 may repress the EMT and decrease radio-resistance of NPC via p-Akt signaling pathway , which may provide a new insight into radio-resistance in NPC.%目的：研究miR-124对鼻咽癌细胞株上皮间充质转化（epithelial- mesenchymal transitions， EMT）和放疗敏感性的影响。方法：转染miR-124 mimic 或inhibitor入鼻咽癌细胞株，划痕实验检测miR-124对鼻咽癌EMT 影响；细胞转染成功24 h 后 X 线照射，检测细胞株中 Caspase-3活性及凋亡细胞比例；EdU 方法检测放疗后细胞增殖能力；Western blot检测信号通路蛋白。结果：划痕实验结果显示 miR-124能抑制鼻咽癌细胞株EMT。 Caspase-3活性检测结果提示，miR-124过表达细胞株接受放疗后 Caspase-3活性显著上升（P ＜0．01）。流式细胞仪结果显示，miR-124能增加细胞株放疗后凋亡。EdU结果显示，miR-124能抑制放疗后细胞的增殖。 Western blot 结果表明过表达 miR-124的细胞株接受放疗后，p-Akt 含量显著下降。结论：miR-124可能通过Akt信号通路抑制鼻咽癌细胞株EMT，增加其放疗敏感性。这为鼻咽放疗增敏提供了新的可行的研究方向。