长链非编码RNA GAS5调节喉癌细胞生物学行为的实验研究

临床肿瘤学杂志 ›› 2018, Vol. 23 ›› Issue (9): 811-816.

长链非编码RNA GAS5调节喉癌细胞生物学行为的实验研究

461000 河南许昌许昌市中心医院耳鼻喉科

收稿日期:2018-04-27 修回日期:2018-07-11 出版日期:2018-09-30 发布日期:2018-11-28

Experimental study of long non-coding RNA GAS5 regulating the biological behavior of laryngeal carcinoma cells

Department of ENT， Xuchang Central Hospital， Xuchang 461000， China

Received:2018-04-27 Revised:2018-07-11 Online:2018-09-30 Published:2018-11-28

摘要/Abstract

摘要： 目的探讨长链非编码RNA GAS5（lncRNA GAS5）对喉癌Hep2细胞增殖和凋亡的影响及可能机制。方法采用实时荧光定量PCR（QPCR）检测喉癌和癌旁正常组织中GAS5的表达，分析其表达与喉癌临床病理特征的关系。Hep2细胞中分别转染pcDNA3.1-GAS5或siGAS5，另分别转染pcDNA3.1-NC或siNC作对照。MTT法检测转染后Hep2细胞增殖活性，流式细胞术检测Hep2细胞周期和凋亡，QPCR法和Western blotting法检测E2F1、BTF3 mRNA和蛋白的表达。结果喉癌组织和癌旁正常组织中GAS5 表达水平分别为0.56±0.10和0.98±0.11，差异有统计学意义（P＜0.05）。GAS5表达与分化程度、TNM分期有关。pcDNA3.1-GAS5组24、48 、72 、96 h Hep2细胞存活率明显低于pcDNA3.1-NC组，差异有统计学意义（P＜0.05）；siGAS5组细胞存活率明显高于siNC组，差异有统计学意义（P＜0.05）。pcDNA3.1-GAS5组细胞G1期细胞比例为（68.14±4.33）％，高于pcDNA3.1-NC组，差异有统计学意义（P＜0.05）。pcDNA3.1-GAS组凋亡率高于pcDNA3.1-NC组，差异有统计学意义（P＜0.05）。pcDNA3.1-GAS5组细胞E2F1、BTF3 mRNA和蛋白的水平均低于pcDNA3.1-NC组，差异具有统计学意义（P＜0.05）；siGAS5组细胞E2F1、BTF3 mRNA和蛋白的水平均高于siNC组，差异具有统计学意义（P＜0.05）。结论 GAS5可负性调控E2F1和BTF3表达，从而参与调控喉癌细胞的增殖和凋亡。

关键词: 喉癌, GAS5, 转录因子E2F1, 通用转录因子3, 增殖, 凋亡

Abstract:

Objective To investigate the effect of long non-coding RNA GAS5 （lncRNA GAS5） on proliferation and apoptosis of laryngeal carcinoma Hep2 cells and its possible mechanism. Methods Real-time fluorescence quantitative PCR （QPCR） was used to detect the expression of GAS5 in laryngeal carcinoma and adjacent normal tissues. The relationship between the expression of GAS5 and the clinicopathological features of laryngeal carcinoma was analyzed. Hep2 cells were transfected with pcDNA3.1-GAS5 or siGAS5， and pcDNA3.1-NC or siNC respectively as control. MTT assay was used to detect the proliferation activity of Hep2 cells after transfection， and Hep2 cell cycle and apoptosis were detected by flow cytometry. QPCR and Western blotting were used to detect the expression of E2F1 and BTF3 mRNA and protein. Results The expression levels of GAS5 in laryngeal carcinoma and adjacent normal tissues were 0.56±0.10 and 0.98±0.11 respectively， and the difference was statistically significant（P＜0.05）. The expression of GAS5 was related to the degree of differentiation and TNM stage. The survival rate of Hep2 cells in pcDNA3.1-GAS5 group at 24， 48， 72 and 96 hours was significantly lower than that in pcDNA3.1-NC group （P＜0.05）， and that in siGAS5 group was significantly higher than that in siNC group （P＜0.05）. The proportion of G1 phase cells transfected with pcDNA3.1-GAS5 was（68.14±4.33）％ higher than that of pcDNA3.1-NC group（P＜0.05）. The apoptotic rate in group pcDNA3.1-GAS5 was higher than that in group pcDNA3.1-NC， and the difference was statistically significant （P＜0.05）. The mRNA and protein levels of E2F1 and BTF3 in pcDNA3.1-GAS5 transfection group were significantly lower than those in pcDNA3.1-NC group （P＜0.05）； the mRNA and protein levels of E2F1 and BTF3 in siGAS5 transfection group were significantly higher than those in siNC group （P＜0.05）. Conclusion GAS5 can negatively regulate the expression of E2F1 and BTF3， and thus participate in the regulation of laryngeal cancer cell proliferation and apoptosis.

Key words: Laryngeal carcinoma, GAS5, Transcription factor E2F1, Basic transcription factor 3, Proliferation, Prognosis

贺松坡, 曹银生, 张晓静.

长链非编码RNA GAS5调节喉癌细胞生物学行为的实验研究 [J]. 临床肿瘤学杂志, 2018, 23(9): 811-816.

HE Songpo, CAO Yinsheng, ZHANG Xiaojing..

Experimental study of long non-coding RNA GAS5 regulating the biological behavior of laryngeal carcinoma cells [J]. Chinese Clinical Oncology, 2018, 23(9): 811-816.

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