沉默长链非编码RNA HOTAIR对舌鳞状细胞癌细胞放射敏感性的影响

Effect of Silencing Long Noncoding RNA HOTAIR on Radiosensitivity of Tongue Squamous Cell Cells

目的:研究下调长链非编码RNA HOTAIR的表达水平对舌鳞状细胞癌细胞株CAL-27和SCC-9放射敏感性的影响。方法:设计3条干扰HOTAIR的序列,通过慢病毒载体转染至CAL-27和SCC-9中,运用实时荧光定量聚合酶链反应(qRT-PCR)验证干扰效率。将细胞分为空白组(CAL-27、SCC-9)、实验组(si-HOTAIR CAL-27、si-HOTAIR SCC-9)、阴性对照组(si-NC CAL-27、si-NC SCC-9),再将以上细胞给予8 Gy电子线照射,MTT法检测各组细胞增殖,划痕实验检测各组细胞迁移,流式细胞实验检测各组细胞凋亡情况。结果:在3条慢病毒载体中,si-2及si-3可下调HOTAIR在CAL-27和SCC-9中的表达(P<0.05),且si-3干扰效率最高(P<0.05),在接受8 Gy电子线照射后,与对照组(si-NC CAL-27、si-NC SCC-9)相比,实验组(si-HOTAIR CAL-27、si-HOTAIR SCC-9)的增殖能力明显减弱(P<0.05);细胞凋亡率增加[(23.87±1.97)%vs.(46.03±2.49)%,(15.99±0.76)%vs.(39.38±2.95)%],迁移率下降[65.85%vs. 58.82%,47.81%vs. 37.78%],差异有统计学意义(P<0.05)。结论:降低HOTAIR的表达水平可提高舌鳞癌细胞CAL-27和SCC-9的放射敏感性。

Objective:To investigate the effect of down-regulated expression of long-chain non-coding RNA HOTAIR on the radiosensitivity of tongue squamous cell lines CAL-27 and SCC-9.Methods:Three HOTAIR interference sequences were designed and transfected into tongue squamous cell cells CAL-27 and SCC-9 by lentiviral vector.The interference efficiency was verified by real-time fluorescence quantitative PCR(qRT-PCR).The cells were divided into blank group(CAL-27,SCC-9),experimental group(si-HOTAIR CAL-27,si-HOTAIR SCC-9),and negative control group(si-NC CAL-27,si-NC SCC-9).The cells were irradiated with 8Gy electron line.MTT assay was used to detect the cell proliferation,scratch assay was used to detect the cell migration,and flow cytometry was used to detect the cell apoptosis.Results:Among three lectin vectors,si-2 and si-3 down-regulated HOTAIR expression in tongue squamous cell cells CAL-27 and SCC-9(P<0.05),and si-3 had the highest interference efficiency(P<0.05).After receiving 8Gy electron line irradiation,compared with the control group(si-NC CAL-27 and si-NC SCC-9),the proliferative ability of the experimental group(si-HOTAIR CAL-27 and SI-NC SCC-9)was significantly decreased(P<0.05),apoptosis rate was increased[(23.87±1.97)%vs.(46.03±2.49)%,(15.99±0.76)%vs.(39.38±2.95)%],and migration rate was decreased[65.85%vs.58.82%,47.81%vs.37.78%].The difference was statistically significant(P<0.05).Conclusion:Reducing the expression level of HOTAIR can increase the radiosensitivity of CAL-27 and SCC-9 of tongue squamous cell.