Chen L, Qiu W F, Cui Z M, Yang H, Tang H W, Luo H
Dongguan Key Laboratory of Environmental Medicine, School of Public Health, Guangdong Medical University, Dongguan 523808, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2022 Oct 20;40(10):721-726. doi: 10.3760/cma.j.cn121094-20210706-00328.
To investigate the cell cycle and apoptosis in hydroquinone (HQ) -induced malignant transformation of TK6 cells and its related regulatory mechanisms. TK6 cells were exposed to 20 μmol/L HQ, 24 h/time, once a week, for 19 weeks as experimental group and TK6 cells treated with phosphate buffer (PBS) for 19 weeks was used as control group from March 2014. In regulatory mechanism research, the cells were divided into four groups: control group, experimental group, control inhibitor group and experimental inhibitor group (inhibitor groups were added 10 μmol/L P600125) . Cell cycle and apoptosis were detected by flow cytometry. The protein expression of cell cycle-related proteins and JNK signaling pathway proteins were detected by Western blot. Flow cytometry showed that compared with control group, the ratio of cells in the G0/G1 phase of the experimental group was significantly decreased (=0.001) , and the ratio of cells in the S phase was significantly increased (=0.002) . Western blotting demonstrated that the protein expressions of p-Rb (Ser780) , E2F1, Cyclin D1, p-p16 (Ser152) , JNK1, p-JNK1 (Thr183/Tyr185) , c-jun, p-c-jun (Ser63) (=0.015, 0.021, 0.001, 0.001, 0.005, 0.001, 0.039, 0.003) were up-regulated, while the protein expressions of Rb (=0.048) and p16 (=0.002) were significantly down-regulated. After exposed to SP600125, compared with experimental group, there were no significant changes in cell cycle distribution (=0.946) and apoptosis rate (=0.923) in experimental inhibitor group. The expression of c-jun (=0.040) protein was down-regulated, while the expression of Rb (=0.027) protein was up-regulated in experimental inhibitor group. In HQ-induced TK6 cells malignant transformation, the cell cycle is arrested in the S phase, and the p16/pRb signaling pathway is inhibited, while the JNK signaling pathway is activated. However, the activated JNK signaling pathway may not be involved in the regulation of cell cycle.
研究对苯二酚（HQ）诱导TK6细胞恶性转化过程中的细胞周期及凋亡情况及其相关调控机制。将TK6细胞暴露于20μmol/L的HQ中，每周1次，每次24小时，共19周作为实验组，自2014年3月起，用磷酸盐缓冲液（PBS）处理19周的TK6细胞作为对照组。在调控机制研究中，将细胞分为四组：对照组、实验组、对照抑制剂组和实验抑制剂组（抑制剂组添加10μmol/L SP600125）。采用流式细胞术检测细胞周期和凋亡情况。通过蛋白质印迹法检测细胞周期相关蛋白和JNK信号通路蛋白的表达。流式细胞术显示，与对照组相比，实验组G0/G1期细胞比例显著降低（=0.001），S期细胞比例显著升高（=0.002）。蛋白质印迹法表明，p-Rb（Ser780）、E2F1、细胞周期蛋白D1、p-p16（Ser152）、JNK1、p-JNK1（Thr183/Tyr185）、c-jun、p-c-jun（Ser63）的蛋白表达上调（=0.015、0.021、0.001、0.001、0.005、0.001、0.039、0.003），而Rb（=0.048）和p16（=0.002）的蛋白表达显著下调。暴露于SP600125后，与实验组相比，实验抑制剂组细胞周期分布（=0.946）和凋亡率（=0.923）无显著变化。实验抑制剂组c-jun（=0.040）蛋白表达下调，而Rb（=0.027）蛋白表达上调。在HQ诱导的TK6细胞恶性转化过程中，细胞周期阻滞于S期，p16/pRb信号通路被抑制，而JNK信号通路被激活。然而，激活的JNK信号通路可能不参与细胞周期的调控。
