Zhou E, Luo L, Ma L J, Yin J, Tan Z Q, Miao G Y, Liu J, Xiao X P
Department of Otorhinolaryngology Head and Neck Surgery, Hunan Provincial People's Hospital, Changsha 410005, China.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2019 Jul 7;54(7):534-539. doi: 10.3760/cma.j.issn.1673-0860.2019.07.010.
To investigate the diagnostic value of serum miRNAlet-7a in laryngeal carcinoma and the effect of let-7a on proliferation and apoptosis of laryngeal carcinoma cells. Real-time quantitative PCR was used to determine the expression level of serum miRNAlet-7a. The miRNA let-7a mimetic was synthesized and transiently transfected into the laryngeal carcinoma Hep-2 cell line by cationic liposome method. The effects of up-regulation of let-7a expression on laryngeal cancer Hep-2 cells were detected by FCM and MTT assays,respectively. The association of let-7a levels with laryngeal cancer and the diagnostic value for laryngeal cancer were analyzed. Measurement data were taken by test or analysis of variance; Counting data were analyzed by χ(2) test and Fisher exact probability method. The receiver operating characteristic curve was used to analyze the diagnostic value of let-7a for laryngeal cancer. The relative expression of serum let-7a in healthy subjects was significantly higher than that in patients with laryngeal cancer (0.931±0.094) (0.380±0.113) (26.507,0.01). The relative expressions of serum let-7a in patients with laryngeal cancer before and after surgery were (0.380±0.113) (0.493±0.164),with significant difference (3.848,0.01).The relative expression of serum let-7a was related to lymph node metastasis (=2.946, 0.01). There was a positive correlation between the relative expression of let-7a in laryngeal carcinoma and that in serum (0.466,0.003). After transfection of let-7a mimics, Hep-2 cells showed an increased significant increase in the expression of let-7a (0.01), proliferation (0.01) and apoptosis (0.01). ROC curve analysis showed that the best critical value for relative expression of let-7a in the diagnosis of laryngeal carcinoma was 0.557 with a sensitivity of 0.794,a specificity of 0.727,an area under curve(AUC) of 0.859,and a 95 of 0.773-0.926. miRNA let-7a can inhibit the proliferation of laryngeal carcinoma Hep-2 cells and promote apoptosis. Serum let-7a is down-regulated in patients with laryngeal cancer and the level of let-7a is related to lymph node metastasis,which would help early diagnosis and postoperative disease monitoring of laryngeal cancer,but further research is needed.
探讨血清微小RNA（miRNA）let-7a在喉癌中的诊断价值及let-7a对喉癌细胞增殖和凋亡的影响。采用实时定量聚合酶链反应（PCR）检测血清miRNA let-7a的表达水平。合成miRNA let-7a模拟物，通过阳离子脂质体法将其瞬时转染至喉癌Hep-2细胞系。分别采用流式细胞术（FCM）和噻唑蓝（MTT）比色法检测上调let-7a表达对喉癌Hep-2细胞的影响。分析let-7a水平与喉癌的相关性及对喉癌的诊断价值。计量资料采用t检验或方差分析；计数资料采用χ²检验和Fisher确切概率法分析。采用受试者工作特征（ROC）曲线分析let-7a对喉癌的诊断价值。健康受试者血清let-7a的相对表达水平显著高于喉癌患者（0.931±0.094） （0.380±0.113）（26.507，P＜0.01）。喉癌患者手术前后血清let-7a的相对表达分别为（0.380±0.113）、（0.493±0.164），差异有统计学意义（3.848，P＜0.01）。血清let-7a的相对表达与淋巴结转移有关（χ²=2.946，P＜0.01）。喉癌组织中let-7a的相对表达与血清中let-7a的相对表达呈正相关（r=0.466，P=0.003）。转染let-7a模拟物后，Hep-2细胞中let-7a的表达显著增加（P＜0.01），细胞增殖受到抑制（P＜0.01），凋亡增加（P＜0.01）。ROC曲线分析显示，let-7a相对表达诊断喉癌的最佳临界值为0.557，灵敏度为0.794，特异度为0.727，曲线下面积（AUC）为0.859，95%置信区间为0.773～0.926。miRNA let-7a可抑制喉癌Hep-2细胞增殖并促进其凋亡。喉癌患者血清let-7a表达下调，且let-7a水平与淋巴结转移有关，这有助于喉癌的早期诊断和术后病情监测，但仍需进一步研究。
