华西口腔医学杂志 ›› 2017, Vol. 35 ›› Issue (1): 57-62.doi: 10.7518/hxkq.2017.01.008

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沉默生物钟基因PER1对口腔鳞状细胞癌细胞中生物钟基因网络的影响

赵钦(), 敖奕然, 杨凯(), 苏小丽, 吕晓强   

  1. 重庆医科大学附属第一医院口腔颌面外科,重庆 400016
  • 收稿日期:2016-07-10 修回日期:2016-09-10 出版日期:2017-02-01 发布日期:2017-02-01
  • 作者简介:

    赵钦,硕士,E-mail:zhaoqinzxc123@163.com

Effect of clock gene PER1 knockdown on clock gene networks in human oral squamous cell carcinoma

Qin Zhao(), Yiran Ao, Kai Yang(), Xiaoli Su, Xiaoqiang. Lü   

  1. Dept. of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Chong-qing Medical University, Chongqing 400016, China
  • Received:2016-07-10 Revised:2016-09-10 Online:2017-02-01 Published:2017-02-01

摘要:

目的 探讨口腔鳞状细胞癌细胞中生物钟基因PER1对生物钟基因网络中其他生物钟基因表达的影响。方法 采用RNA干扰技术沉默人口腔鳞状细胞癌SCC15细胞中PER1基因,应用流式细胞仪检测沉默后细胞的增殖和凋亡水平,实时荧光定量聚合酶链式反应检测生物钟基因CLOCK、BMAL1、PER1、PER2、PER3、DEC1、DEC2、CRY1、CRY2、TIM、CKIE、RORA、NPAS2、REV-ERBA mRNA的表达情况。结果 PER1基因沉默后,SCC15细胞增殖指数上升,凋亡指数下降(P<0.05);PER1、PER2、DEC1、DEC2、CRY1、CRY2和NPAS2 mRNA的表达水平降低(P<0.05),PER3、TIM、RORA和REV-ERBA mRNA的表达水平升高(P<0.05),CLOCK、BMAL1和CKIE mRNA的表达水平无统计学改变(P>0.05)。结论 生物钟基因PER1能够调控生物钟基因网络中众多其他生物钟基因PER2、DEC1、DEC2、CRY1、CRY2、NPAS2、PER3、TIM、RORA和REV-ERBA的表达,PER1在生物钟基因网络中具有重要调控作用。

关键词: 生物钟, 鳞状细胞癌, 口腔, 基因

Abstract:

Objective This study investigated the effect of clock gene PER1 on the expression levels of other clock genes in clock gene networks in oral squamous cell carcinoma cells. Methods We used RNA interference mediated by short hairpin RNAs (shRNAs) to effectively knock down PER1 in SCC15 human oral squamous cell carcinoma cells. Flow cytometry was used to detect the degree of proliferation and apoptosis of the cells after PER1 knockdown, and quantitative real-time PCR was used to detect the mRNA expression levels of the clock genes CLOCK, BMAL1, PER1, PER2, PER3, DEC1, DEC2, CRY1, CRY2, TIM, CKIE, RORA, NPAS2, and REV-ERBA. Results The proliferation index of SCC15 cells increased significantly while the apoptotic index decreased significantly after PER1 knockdown (P<0.05). The mRNA expression levels of PER1, PER2, DEC1, DEC2, CRY1, CRY2, and NPAS2 markedly decreased (P<0.05) while those of PER3, TIM, RORA, and REV-ERBA markedly increased (P<0.05). By contrast, no obvious changes were observed in the mRNA expression levels of CLOCK, BMAL1, and CKIE (P>0.05). Conclusion The clock gene PER1 can regulate the expression levels of other clock genes in the clock gene networks; these genes include PER2, DEC1, DEC2, CRY1, CRY2, NPAS2, PER3, TIM, RORA, and REV-ERBA. PER1 gene thus plays an important role in the regulation of clock gene networks.

Key words: circadian clock, squamous cell carcinoma, oral cavity, gene

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