p16/pRb和JNK信号通路在氢醌诱导TK6细胞恶性转化中的作用  

Effects of p16/pRb and JNK signaling pathways in hydroquinone-induced malignant transformation of TK6 cells

作　　者：陈林 邱伟峰 崔哲明 杨慧 唐焕文 罗皓 Chen Lin;Qiu Weifeng;Cui Zheming;Yang Hui;Tang Huanwen;Luo Hao(Dongguan Key Laboratory of Environmental Medicine,School of Public Health,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]广东医科大学公共卫生学院,东莞市环境医学重点实验室,东莞523808

出　　处：《中华劳动卫生职业病杂志》2022年第10期721-726,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金青年项目(81803278);广东省自然科学基金面上项目(2018A030313580、2021A1515010950);广东省医学科学技术研究基金项目(A2018197、A2019404);广东医科大学学科建设项目(4SG21003G)。

摘　　要：目的探讨氢醌(HQ)诱导的人淋巴母细胞(TK6)恶性转化细胞周期和凋亡情况及其相关的调控机制。方法于2014年3月,以20μmol/L HQ染毒TK6细胞,每次染毒时间为24 h,每周1次,处理19周,设为实验组,同时以磷酸盐缓冲液(PBS)处理19周的TK6细胞为对照组;在调控机制的探讨中将细胞分为4组,即对照组、实验组、对照抑制剂组、实验抑制剂组(加入10μmol/L P600125)。采用流式细胞术检测细胞周期和凋亡的情况;Western成blot检测细胞周期相关蛋白和JNK信号通路蛋白的表达水平。结果与对照组比较,实验组G0/G1期细胞所占比例明显降低(P=0.001),S期细胞所占比例明显增加(P=0.002);实验组细胞p-Rb(Ser780)、E2F1、Cyclin D1、p-p16(Ser152)、JNK1、p-JNK1(Thr183/Tyr185)、c-jun、p-c-jun(Ser63)蛋白表达均上调(P=0.015、0.021、0.001、0.001、0.005、0.001、0.039、0.003),而Rb、p16蛋白表达明显下调(P=0.048、0.002);加入SP600125抑制JNK信号通路后,与实验组比较,实验抑制剂组细胞周期分布和细胞凋亡率均无明显变化,差异无统计学意义(P=0.946、0.923);c-jun蛋白表达下调,Rb蛋白表达上调,差异有统计学意义(P=0.040、0.027)。结论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,但被激活的JNK信号通路可能不参与调控HQ诱导的TK6细胞恶性转化过程中的细胞周期改变进程。Objective To investigate the cell cycle and apoptosis in hydroquinone(HQ)-induced malignant transformation of TK6 cells and its related regulatory mechanisms.Methods TK6 cells were exposed to 20μmol/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 10μmol/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.Results Flow cytometry showed that compared with control group,the ratio of cells in the G0/G1 phase of the experimental group was significantly decreased(P=0.001),and the ratio of cells in the S phase was significantly increased(P=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)(P=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(P=0.048)and p16(P=0.002)were significantly down-regulated.After exposed to SP600125,compared with experimental group,there were no significant changes in cell cycle distribution(P=0.946)and apoptosis rate(P=0.923)in experimental inhibitor group.The expression of c-jun(P=0.040)protein was down-regulated,while the expression of Rb(P=0.027)protein was up-regulated in experimental inhibitor group.Conclusion 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.

关 键 词：氢醌类 细胞周期 细胞凋亡 恶性转化 JNK信号通路 p16/pRb信息通路 

分 类 号：R114[医药卫生—卫生毒理学]