SBF2-AS1在食管鳞癌中的表达变化及其对肿瘤细胞KYSE410增殖、放疗敏感性的影响

Changes of SBF2-AS1 expression in esophageal squamous cell carcinoma and their effects on prolifer⁃ation and radiosensitivity of tumor cells KYSE410

目的观察长链非编码RNA SBF2-AS1在食管鳞癌中的表达变化及其对食管鳞癌细胞增殖、放疗敏感性的影响,并探索可能的机制。方法采用qRT-PCR法检测食管鳞癌组织和食管鳞癌细胞KYSE410中的SBF2-AS1,分析食管鳞癌组织中SBF2-AS1表达与肿瘤临床病理参数的关系。培养KYSE410细胞并分为siRNA-SBF2-AS1组、NC组,分别转染siRNA-SBF2-AS1、NC,分别于转染24、48、72、96 h后检测细胞增殖能力。对两组细胞给予X线照射,测算不同照射条件下的细胞增殖率和凋亡率;绘制单击多靶模型拟合细胞存活曲线得出D0(终斜率的倒数)、Dq(准阈剂量)、SF2(离体肿瘤培养细胞经2 Gy照射后的SF),计算放射增敏比;检测细胞中的E2F转录因子1(E2F1)蛋白。结果食管鳞癌组织和细胞中SBF2-AS1表达高于癌旁正常组织和食管正常上皮细胞(P均<0.05)。肿瘤直径>5 cm者、T3~T4期者SBF2-AS1相对表达量分别高于直径≤5 cm者、T1~T2期者(P均<0.05)。转染24、48、72、96 h后siRNASBF2-AS1组细胞增殖能力低于NC组(P均<0.05)。不同X线照射条件的siRNA-SBF2-AS1组细胞增殖率、E2F1蛋白表达均低于NC组,细胞凋亡率高于NC组;照射后细胞增殖率和E2F1蛋白表达低于未照射细胞,细胞凋亡率高于未照射细胞(P均<0.05)。siRNA-SBF2-AS1组细胞D0、Dq、SF2低于NC组(P均<0.05),放射增敏比为2.00±0.10。结论SBF2-AS1在食管癌组织和细胞中高表达;下调SBF2-AS1表达可抑制食管鳞癌细胞增殖,提高放疗敏感性,其机制可能与调控E2F1蛋白表达有关。

Objective To observe the expression changes of long-chain non-coding RNA SBF2-AS1 in the esophageal squamous cell carcinoma and their effects on cell proliferation and radiation sensitivity of esophageal squamous cell carcinoma cells and to investigate the possible mechanism.Methods The qRT-PCR method was used to detect the SBF2-AS1 in the esophageal squamous cell carcinoma tissues and esophageal squamous cell carcinoma cell line KYSE410 and to analyze the relationship between SBF2-AS1 expression and tumor clinicopathological parameters in the esophageal squamous cell carcinoma tissues.KYSE410 cells were cultured and divided into the siRNA-SBF2-AS1 group and NC group,which were treasfected with siRNA-SBF2-AS1 and NC,respectively;the cell proliferation ability was detected at 24,48,72,and 96 h after transfection.The cells of the two groups were subjected to X-ray irradiation,and we calculated the cell proliferation rate and apoptosis rate under different irradiation conditions;the D0(reciprocal of final slope),Dq(quasi-threshold dose),and SF2(SF of isolated tumor cultured cells after 2 Gy irradiation)were obtained by drawing the cell survival curve fitted by multiple target models,and then we calculated the radiosensitization ratio and detected the E2F transcription factor 1(E2F1)protein in the cells.Results The expression of SBF2-AS1 in the esophageal squamous cell carcinoma was higher than that in the adjacent normal tissues and esophageal normal epithelial cells(P<0.05).The relative expression of SBF2-AS1 in patients with tumor diameter>5 cm,in the T3-T4 stage was higher than that of patients with diameter<5 cm and in the T1-T2 stage(all P<0.05).The proliferation abilities of the siRNA-SBF2-AS1 group after transfection for 24,48,72,and 96 h were lower than those the NC group(all P<0.05).The cell proliferation rate and E2F1 protein expression in the siRNA-SBF2-AS1 group with different X-ray irradiation conditions were lower than those in the NC group,and the cell apoptosis rate was higher than that in the NC group;the cell proliferation rate and E2F1 protein expression were lower in the cells after irradiation than in the cells without irradiation,but the apoptosis rate was higher(all P<0.05).The cell D0,Dq,and SF2 of the siRNA-SBF2-AS1 group were lower than those of the NC group(all P<0.05),and the radiosensitization ratio was 2.00±0.10.Conclusion SBF2-AS1 is highly expressed in the esophageal cancer tissues and cells,and down-regulation of SBF2-AS1 expression can inhibit the proliferation of esophageal squamous cell carcinoma cells and improve the sensitivity of radiotherapy;the mechanism may be related to the regulation of E2F1 protein expression.