上调HuR基因对食管鳞癌细胞Kyse450放射敏感性的影响

Effect of upregulated HuR gene on radiosensitivity of esophageal squamous cell carcinoma cell Kyse450

目的研究HuR基因上调对于食管鳞癌细胞Kyse450放射敏感性的影响。方法慢病毒上调Kyse450细胞HuR基因,同时选取X射线6 Gy照射剂量作为干预条件。采用Western blot和qPCR技术分别检测Kyse450转染后的蛋白和RNA表达情况;CCK8试剂盒检测细胞的增殖速率;克隆形成试验检测细胞克隆形成能力;伤口愈合试验和Transwell试验检测细胞迁移能力的改变。结果CCK8试验显示HuR基因上调后细胞的增殖能力增强,放射后这种增强趋势更加明显;平板克隆试验显示随着放射剂量的增加,两组细胞的克隆形成率都随之降低,但是过表达组的克隆形成数多于对照组;伤口愈合试验以及Transwell试验表明过表达组的伤口愈合速率和迁移能力高于对照组,放射治疗后这种差异更显著;Western blot显示放射治疗后24 h过表达组细胞内MMP9、MMP2水平高于对照组。结论HuR上调会增强食管鳞癌细胞增殖、克隆、迁移能力,降低其放射敏感性。

Objective To evaluate the effect of upregulation of HuR gene on the radiosensitivity of esophageal squamous cell carcinoma cell Kyse450.Methods The HuR gene of Kyse450 cells was upregulated by lentivirus.At the same time,X-ray irradiation at a dose of 6Gy was selected as the intervention condition.Western blot and qPCR were used to detect the expression levels of protein and RNA after Kyse450 transfection,respectively.CCK8 kit was employed to determine the cell proliferation rate.Clone formation assay was adopted to evaluate the ability of cell clone formation.Wound healing experiment and the Transwell test were performed to detect changes in cell migration.Results CCK8 assay showed that the proliferation ability of cells was enhanced after upregulation of HuR gene,and this enhancement trend was more obvious after radiation.The plate cloning experiment showed that with the increase of radiation dose,the clone formation rates were decreased in both groups,but the clone formation rate in the overexpression group was higher than that in the control group.Wound healing experiment and Transwell test demonstrated that the wound healing rate and migration ability in the overexpression group were higher than those in the control group,and the difference was more significant after radiotherapy.Western blot showed that the levels of MMP9 and MMP2 at 24h after radiotherapy in the overexpression group were higher than those in the control group.Conclusion The upregulation of HuR can enhance the proliferation,cloning,migration capabilities and decrease the radiosensitivity of esophageal squamous cell carcinoma cells.