lncRNA SBF2-AS1低表达提高食管鳞癌的放射敏感度

Low Expression of LncRNASBF2-AS1 Increases Radiosensitivity in Esophageal Squamous Cell Carcinoma

目的探究长链非编码RNA SBF2-AS沉默后对食管鳞癌细胞放射敏感度的影响。方法用实时荧光定量PCR(qRT-PCR)检测TE-13和HET-1A细胞中SBF2-AS1的表达水平。将TE-13细胞分为si-NC组、si-SBF2-AS1组、IR+si-NC组、IR+si-SBF2-AS1组。四甲基偶氮唑蓝(MTT)和EDU(胸腺嘧啶核苷酸类似物)检测各组细胞增殖。流式细胞术检测各组细胞凋亡。蛋白质印记(Western blot)法检测各组细胞中E2F1蛋白的变化。结果TE-13细胞中SBF2-AS1的表达水平明显高于HET-1A(P<0.05)。与IR+si-NC组比较,IR+si-SBF2-AS1组细胞存活分数呈剂量依赖性显著下降(P<0.05)。与si-NC组比较,IR+si-NC组、si-SBF2-AS1组和IR+si-SBF2-AS1组在48h和72h的A值显著降低(P<0.05)。与IR+si-NC组比较,IR+si-SBF2-AS1组48h和72h的A值显著降低(P<0.05)。与si-SBF2-AS1组比较,IR+si-SBF2-AS1组48h和72h的A值显著降低(P<0.05)。与si-NC组比较,IR+si-NC组、si-SBF2-AS1组和IR+si-SBF2-AS1组增殖率显著降低(P<0.05)。与IR+si-NC组比较,IR+si-SBF2-AS1组增殖率显著降低(P<0.05)。与si-SBF2-AS1组比较,IR+si-SBF2-AS1组增殖率显著降低(P<0.05)。与si-NC组比较,IR+si-NC组、si-SBF2-AS1组和IR+si-SBF2-AS1组的细胞凋亡率显著升高(P<0.05)。与IR+si-NC组比较,IR+si-SBF2-AS1组的细胞凋亡率显著升高(P<0.05)。与si-SBF2-AS1组比较,IR+si-SBF2-AS1组的细胞凋亡率显著升高(P<0.05)。与IR+si-NC组比较,IR+si-SBF2-AS1组E2F1蛋白的表达量显著下降(P<0.05)。结论敲除SBF2-AS1可以通过抑制TE-13细胞增殖,促进其凋亡,来增强其对放射治疗的敏感度,为提高食管鳞癌放射治疗疗效提供一个新的靶点。

Objective To investigate the biological role of SBF2-AS1 in promoting radiotherapy resistance of esophageal squamous cell carcinoma.Methods Detection of SBF2-AS1 expression in TE-13 and HET-1A cells by real-time fluorescence quantitative PCR(qRT-PCR)was performed.TE-13 cells were divided into si-NC group,si-SBF2-AS1 group,IR+si-NC group and IR+si-SBF2-AS1 group.Cell proliferation was detected by MTT and EDU.The apoptosis of each group was detected by flow cytometry,and the change of E2F1 protein in each group was detected by westernblot.Results The expression level of SBF2-AS1 in TE-13 cells was significantly higher than that in HET-1A group(P<0.05).Compared with IR+si-NC group,the cell survival fraction of IR+si-SBF2-AS1 group decreased significantly in a dose-dependent manner(P<0.05).Compared with si-NC group,the A values of IR+si-NC group,si-SBF2-AS1 group and IR+si-SBF2-AS1 group were significantly lower at 48h and 72h(P<0.05).Compared with IR+si-NC group,the A values of IR+si-SBF2-AS1 group at 48h and 72h were significantly lower(P<0.05).Compared with si-SBF2-AS1 group,the A values of IR+si-SBF2-AS1 group at 48h and 72h were significantly lower(P<0.05).The proliferation rate of IR+si-NC group,si-SBF2-AS1 group and IR+si-SBF2-AS1 cell group was significantly lower than that of si-NC group(P<0.05),that of IR+si-SBF2-AS1 group was significantly lower than that of IR+si-NC group(P<0.05),and that of IR+si-SBF2-AS1 group was significantly lower than that of si-SBF2-AS1 group(P<0.05).Compared with si-NC group,the apoptosis rate of IR+si-NC group,si-SBF2-AS1 group and IR+si-SBF2-AS1 group was significantly higher(P<0.05),that of IR+si-SBF2-AS1 group was significantly higher than that of IR+si-NC group(P<0.05),and that of IR+si-SBF2-AS1 group was significantly higher than that of si-SBF2-AS1 group(P<0.05).Compared with IR+si-NC group,the expression of E2F1 protein in IR+si-SBF2-AS1 group decreased significantly(P<0.05).Couclusion Knockout of SBF2-AS1 can enhance the sensitivity of TE-13 cells to radiotherapy by inhibiting proliferation and promoting apoptosis,which provides a new target for improving the efficacy of radiotherapy for esophageal squamous cell carcinoma.