广西医学

目的探讨长链非编码RNA（lncRNA）核旁斑富集转录本1（NEAT1）在鼻咽癌中的表达情况及其对鼻咽癌放疗敏感性的影响。方法（1）使用UALCAN数据库分析NEAT1在头颈部鳞状细胞癌中的表达情况及其与患者临床分期、预后的关系。（2）利用实时荧光定量PCR检测鼻咽癌组织及正常鼻咽黏膜上皮、人永生化鼻咽上皮细胞NP⁃69及多株鼻咽癌细胞（CNE⁃1、CNE⁃2、CNE⁃2R、C666⁃1、C666⁃1R）中NEAT1的表达情况。（3）使用RNA干扰技术构建NEAT1沉默的CNE⁃2R细胞［NEAT1⁃短发夹RNA（shRNA）组］，并设立对照组（未转染的CNE⁃2R细胞）、NEAT1⁃NC组（转染对照序列的CNE⁃2R细胞）。给予不同放射剂量X线照射后，检测3组CNE⁃2R细胞的存活率、凋亡率、克隆形成率及放射生物学参数。结果（1）NEAT1在头颈部鳞状细胞癌组织中呈高表达，且表达水平随着头颈部鳞状细胞癌分期的增加呈升高趋势（P<0.05），但不同NEAT1表达水平患者的总生存率差异无统计学意义（P > 0.05）。（2）相较于正常鼻咽黏膜上皮组织，鼻咽癌组织中NEAT1表达上调（P<0.05）。相较于NP⁃69细胞，NEAT1在鼻咽癌细胞中的表达上调（P<0.05），其中CNE⁃2R细胞中的表达量最高（P<0.05）；放射抗拒性细胞CNE⁃2R、C666⁃1R中NEAT1的表达量分别高于放疗敏感性细胞CNE⁃2、C666⁃1（P<0.05）。（3）在接受不同剂量X射线照射后，与对照组及NEAT1⁃NC 组比较，NEAT1⁃shRNA 组CNE⁃2R细胞存活率、克隆形成率及放射生物学参数降低，凋亡率升高（P<0.05），但对照组与NEAT1⁃NC 组上述指标差异无统计学意义（P>0.05）。结论 lncRNA NEAT1在鼻咽癌组织和细胞中呈高表达，与肿瘤分期密切相关，沉默NEAT1的表达可增加鼻咽癌的放疗敏感性。

Objective To explore the expression of long non⁃coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) in nasopharyngeal carcinoma and its influence on radiotherapy sensitivity of nasopharyngeal carcinoma. Methods (1) NEAT1 expression in head and neck squamous cell carcinoma and its relation with clinical stage and prognosis of patients were analyzed through using the UALCAN database. (2) The real⁃time fluorescent quantitative PCR was used to detect NEAT1 expression in nasopharyngeal cancerous tissues and normal nasopharyngeal mucosal epithelium, in human immortalized nasopharyngeal epithelial cells NP⁃69, and in multiple strains of nasopharyngeal carcinoma cells (CNE⁃1, CNE⁃2, CNE⁃2R, C666⁃1, and C666⁃1R). (3) The RNA interference technique was used to construct NEAT1⁃silenced CNE⁃2R cells (the NEAT1⁃shot hairpin RNA ［shRNA］ group), and control group (non⁃transfection CNE⁃2R cells), and NEAT1⁃NC group (CNE⁃2R cells transfected with the control sequence) were set. After X⁃ray irradiation with different radian doses, the survival rate, apoptosis rate, cloning formation rate, and radiobiological parameters of CNE⁃2R cells were detected in the 3 groups. Results (1) NEAT1 was highly expressed in tissues of head and neck squamous cell carcinoma, and its expression was elevated with the progression of stage of head and neck squamous cell carcinoma (P<0.05), but there was no statistically significant difference in the total survival rate between patients with different NEAT1 expressions (P>0.05). (2) Compared with tissues of normal nasopharyngeal mucosal epithelium, NEAT1 expression was up⁃regulated in nasopharyngeal cancerous tissues (P<0.05). Compared with NP⁃69 cells, NEAT1 expression was up⁃regulated in nasopharyngeal cancerous tissues (P<0.05), therein expression in CNE⁃2R cells was the highest (P<0.05). NEAT1 expressions in radioresistant cells CNE⁃2R and C666⁃1R were higher than those in radiotherapy sensitivity cells CNE⁃2 and C666⁃1, respectively (P<0.05). (3) After receiving X⁃ray irradiation with different radiant doses, compared with the control group and the NEAT1⁃NC group, the NEAT1⁃shRNA group yielded decreased survival rate, cloning formation rate and radiobiological parameters of CNE⁃2R cells, whereas an elevated apoptosis rate of CNE⁃2R cells (P<0.05), but there was no statistically significant difference in the aforementioned indices between the control group and the NEAT1⁃NC group (P>0.05). Conclusion The expression of lncRNA NEAT1 is high in nasopharyngeal cancerous tissues and cells, and lncRNA NEAT1 is closely related to tumor stage. Silencing NEAT1 expression can increase radiotherapy sensitivity of nasopharyngeal carcinoma.