miR-377-5p通过抑制AKT1/GSK-3β通路增强食管癌细胞TE-1的放射敏感性

目的 探究miR-377-5p对食管癌细胞TE-1放射敏感性的影响及机制。方法 TE-1细胞转染miR-377-5pmimic和miR-377-5pmimic NC构建过表达miR-377-5p细胞。对转染后的TE-1细胞照射后采用平板集落形成实验检测细胞放射生物学参数(D0、Dq、SF2),Transwell小室法检测细胞侵袭能力,细胞划痕实验检测细胞迁移能力,CCK8法检测细胞活力,流式细胞术检测细胞凋亡和细胞周期,免疫印迹检测AKT1和GSK-3β磷酸化水平。结果 2、4、6、8Gy照射的集落形成率均显著下降(P<0.05),且在同一剂量下miR-377-5pmimic组细胞集落形成率均显著低于miR-377-5pmimic NC组(P<0.05)。相比于miR-377-5pmimic NC组,miR-377-5pmimic组D0、Dq、SF2均显著下降(P<0.05),放射增敏比为1.34(D0值比);0 Gy照射后细胞侵袭、迁移、增殖能力显著下降,细胞凋亡水平显著上升,细胞周期被阻滞于G1期,AKT1和GSK-3β磷酸化水平显著下降(P<0.05);4 Gy照射后细胞侵袭、迁移、增殖能力下降,细胞凋亡水平显著上升,G1期显著延长,AKT1和GSK-3β磷酸化水平亦显著下降(均P<0.001)。结论 miR-377-5p能够增加食管癌细胞TE-1的放射敏感性,其机制可能是抑制AKT1/GSK-3β信号通路。

miR-377-5p enhances radiosensitivity of esophageal cancer cell line TE-1 by inhibiting AKT1/GSK-3β signaling pathway

Objective To investigate the effect and mechanism of miR-377-5p on the radiosensitivity of esophageal cancer cell line TE-1. Methods Esophageal cancer cell line TE-1 was transfected with miR-377-5p mimic and miR-377-5p mimic NC to construct the TE-1 cells overexpressing miR-377-5p. After the transfected TE-1 cells were exposed to ionizing radiation, radiobiological parameters(D0,Dq,SF2) were detected by plate colony formation assay. Cell invasion ability was assessed by Transwell chamber assay, Cell migration ability was evaluated by cell scratch assay, Cell viability was assessed by CCK8 assay. Cell apoptosis and cell cycle were detected by flow cytometry, Western blot. The phosphorylation levels of AKT1 and GSK-3βwere measured by Western blot. Results At the doses of 2, 4, 6, and 8 Gy, the colony formation rate in each group was significantly decreased (all P<0.05), and the colony formation rate at the same irradiation dose in the miR-377-5p mimic group was significantly lower than that in the miR-377-5p mimic NC group (P<0.05). Compared with the miR-377-5pmimic NC group, the D0, Dq and SF2 at 2Gy were decreased significantly in the miR-377-5pmimic group (all P<0.05), and the radiosensitization ratio(D0 ratio) was 1.34. After 0 Gy ionizing radiation, the invasion, migration and proliferation abilities of TE-1 cells were significantly decreased, the level of cell apoptosis was significantly increased, the cell cycle was arrested in G1 phase, and the phosphorylation levels of AKT1 and GSK-3β were significantly decreased in the miR-377-5pmimic group (all P<0.05). Following 4 Gy irradiation, cell invasion, migration, proliferation abilities were decreased, the level of cell apoptosis was increased significantly, the G1 phase was significantly extended and the phosphorylation levels of AKT1 and GSK-3β were also decreased significantly in the miR-377-5pmimic group (all P<0.001). Conclusion miR-377-5p can increase the radiosensitivity of esophageal cancer cell line TE-1, which may be due to the inhibition of the AKT1/GSK-3β signaling pathway.