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[生物钟基因PER1敲低对人口腔鳞状细胞癌生物钟基因网络的影响]

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

作者信息

Qin Zhao, Yiran Ao, Kai Yang, Xiaoli Su, Xiaoqiang Lü

机构信息

Dept. of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2017 Feb 1;35(1):57-62. doi: 10.7518/hxkq.2017.01.008.

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).

CONCLUSIONS: 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.
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摘要

目的

本研究调查了时钟基因PER1对口腔鳞状细胞癌细胞时钟基因网络中其他时钟基因表达水平的影响。

方法

我们使用短发夹RNA(shRNA)介导的RNA干扰,有效敲低SCC15人口腔鳞状细胞癌细胞中的PER1。采用流式细胞术检测PER1敲低后细胞的增殖和凋亡程度,采用定量实时PCR检测时钟基因CLOCK、BMAL1、PER1、PER2、PER3、DEC1、DEC2、CRY1、CRY2、TIM、CKIE、RORA、NPAS2和REV-ERBA的mRNA表达水平。

结果

PER1敲低后,SCC15细胞的增殖指数显著增加,而凋亡指数显著降低(P<0.05)。PER1、PER2、DEC1、DEC2、CRY1、CRY2和NPAS2的mRNA表达水平显著降低(P<0.05),而PER3、TIM、RORA和REV-ERBA的mRNA表达水平显著增加(P<0.05)。相比之下,CLOCK、BMAL1和CKIE的mRNA表达水平未观察到明显变化(P>0.05)。

结论

时钟基因PER1可以调节时钟基因网络中其他时钟基因的表达水平;这些基因包括PER2、DEC1、DEC2、CRY1、CRY2、NPAS2、PER3、TIM、RORA和REV-ERBA。因此,PER1基因在时钟基因网络的调节中起重要作用。

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