白血病相关长链非编码RNA在氢醌诱导TK6恶性转化细胞中的表达及其调控机制  

Expression of leukemia-related lncRNA in hydroquinone-induced TK6 malignant transformed cells and its related regulatory mechanisms

作　　者：袁倩 张海桥 凌晓璇 黄海燕 郑冬燕 李洁优 钟柏森 刘林华 YUAN Qian;ZHANG Hai-qiao;LING Xiao-xuan;HUANG Hai-yan;ZHENG Dong-yan;LI Jie-you;ZHONG Bai-sen;LIU Lin-hua(School of Public Health, Guangdong Medical University, Dongguan, Guangdong 523808,China)

机构地区：[1]广东医科大学公共卫生学院,广东东莞523808 [2]东莞市环境医学重点研究室 [3]广东医科大学公共卫生学院实验教学中心 [4]广东医科大学劳动卫生与环境卫生学教研室

出　　处：《环境与健康杂志》2018年第5期393-397,共5页Journal of Environment and Health

基　　金：国家自然科学基金(81202231);广东省医学科学技术研究基金(A2018225);广东省教育厅青年创新人才项目(2014KQNCX102);广东医科大学校级大学生创新创业训练计划项目(GDMU2017025;2016ZZDG004);广东医科大学博士科研启动项目(B2017021)

摘　　要：目的探讨氢醌(hydroquinone,HQ)对白血病相关长链非编码RNA(lncRNA)的表达影响及其相关的调控机制。方法以磷酸盐缓冲液(PBS)处理的TK6细胞为PBS对照组,以10.0μmol/L HQ诱导的TK6恶性转化细胞为HQ染毒组;在调控机制探讨中以10.0μmol/L HQ诱导的TK6恶性转化细胞为对照,同时设10.0μmol/L HQ+5μmol/L 5-氮杂胞苷(5-Aza C,DNA甲基转移酶酶抑制剂)组和10.0μmol/L HQ+200 nmol/L曲古抑菌素A(TSA,蛋白去乙酰化酶抑制剂)组。以反转录实时荧光定量-聚合酶链反应(q RT-PCR)检测白血病相关lncRNA表达量以及Tet1、Tet2、Tet3的mRNA表达量。以蛋白免疫印迹检测Tet1、Tet2蛋白表达水平。结果与PBS对照组相比,HQ诱导的TK6恶性转化细胞中lncRNA HOTAIRM1、lncRNA BCO35666和lncRNA NEAT1表达下降,lncRNA HOTAIRM1表达下降最为显著,lncRNA NELT1、lncRNA ANKR036BP1、lncRNA LUNAR1、lncRNA SENCR和lncRNA UCA1表达上升,UCA1的表达升高最为明显;Tet1、Tet2、Tet3的mRNA表达下降,Tet2、Tet3的下降均有统计学意义(P<0.05),Tet1、Tet2的蛋白表达下降。与HQ染毒组比较,经5-Aza C与TSA处理的HQ诱导的TK6恶性转化细胞lncRNA HOTAIRM1和lncRNA NEAT1表达上升,而lncRNA BCO35666表达下降;Tet1、Tet2、Tet3 mRNA在HQ+5-Aza C处理组中表达均升高,Tet1、Tet2升高均有统计学意义(P<0.05),而在HQ+TSA处理组中Tet1表达升高(P<0.05),Tet3表达下降(P<0.05);Tet1、Tet2的蛋白表达与mRNA结果一致。结论在HQ诱导的TK6恶性转化细胞中,DNA甲基化与DNA去甲基化可能相互作用共同调控lncRNA HOTAIRM1、lncRNA NEAT1的表达。Objective To investigate the effect of hydroquinone（HQ） on the expression of leukemia-associated lncRNA and its related regulatory mechanism. Methods TK6 cells treated with phosphate buffered saline（PBS） were used as the control group and TK6 malignant transformed cells induced by 10.0 μmol/L HQ were used as the HQ exposure group. In regulatory mechanisms research, HQ-induced TK6 malignant transformed cells were served as the control group, HQ-induced TK6 malignant transformed cells individually treated by DNA methyltransferase enzyme inhibitors（5 μmol/L 5-Aza C） or histone deacetylase inhibitors（200 nmol/L TSA） were used as HQ＋5-Aza C group and HQ＋ TSA group respectively. q RT-PCR was used to detect leukemia-related lncRNA expression and Tet1, Tet2 and Tet3 mRNA expression levels. The protein expression of Tet1 and Tet2 were detected by Western blot. Results Compared with PBS group, the expression of lncRNA HOTAIRM1, lncRNA NEAT1 and lncRNA BCO35666 were decreased in the HQ-induced malignant transformed TK6 cells, especially for lncRNA HOTAIRM1（P〈0.05）; While the expression of lncRNA NELT1, lncRNA ANKR036 BP1, lncRNA LUNAR1, lncRNA SENCR,lncRNA UCA1 were increased, especially for lncRNA UCA1（P〈0.05）; The mRNA expression of Tet1, Tet2 and Tet3 decreased,especially for Tet2 and Tet3（P〈0.05）; The expression of Tet1 and Tet2 protein decreased also. Compared with HQ exposure group, the expression of lncRNA HOTAIRM1 and lncRNA NEAT1 increased and the expression of lncRNA BCO35666 decreased in HQ ＋5-Aza C group and HQ ＋TSA group; The mRNA expression of Tet1, Tet2 and Tet3 increased in HQ ＋5-Aza C group,especially for Tet1 and Tet2（P〈0.05）; Tet1 expression also increased and Tet3 decraesed in HQ ＋5 TSA group; The trends of protein expressions of Tet1 and Tet2 were similar with their mRNA expression. Conclusion In HQ-induced TK6 malignant transformed cells, DNA methylation may interact with DNA demethylation to regulate the expression of lncRNA HOTAIRM1 and lncRNA NEA

关 键 词：氢醌 长链非编码RNA DNA甲基化 DNA去甲基化 恶性转化 白血病 

分 类 号：R994.6[医药卫生—毒理学]