对苯二酚转化细胞来源的外泌体miR-1246通过靶向TK6细胞中的细胞周期蛋白G2驱动S期积累停滞。
Exosomal derived miR-1246 from hydroquinone-transformed cells drives S phase accumulation arrest by targeting cyclin G2 in TK6 cells.
作者信息
Chen Yuting, Chen Lin, Zhu Shiheng, Yang Hui, Ye Zhongming, Wang Huanhuan, Wu Haipeng, Wu Yao, Sun Qian, Liu Xiaoshan, Liang Hairong, Tang Huanwen
机构信息
The First Dongguan Affiliated Hospital, Guangdong Medical University, Dongguan, 523808, China; Dongguan Key Laboratory of Environmental Medicine, School of Public Health, Guangdong Medical University, Dongguan, 523808, China.
Dongguan Key Laboratory of Environmental Medicine, School of Public Health, Guangdong Medical University, Dongguan, 523808, China.
出版信息
Chem Biol Interact. 2024 Jan 5;387:110809. doi: 10.1016/j.cbi.2023.110809. Epub 2023 Nov 23.
BACKGROUND
Hydroquinone (HQ), a major metabolite of benzene and known hematotoxic carcinogen. MicroRNA 1246 (miR-1246), an oncogene, regulates target genes in carcinogenesis including leukemia. This study investigates the impact of exosomal derived miR-1246 from HQ-transformed (HQ19) cells on cell-to-cell communication in recipient TK6 cells.
METHODS
RNA sequencing was used to identify differentially expressed exosomal miRNAs in HQ19 cells and its phosphate buffered solution control cells (PBS19), which were then confirmed using qRT-PCR. The impact of exosomal miR-1246 derived from HQ-transformed cells on cell cycle distribution was investigated in recipient TK6 cells.
RESULTS
RNA sequencing analysis revealed that 34 exosomal miRNAs were upregulated and 158 miRNAs were downregulated in HQ19 cells compared with PBS19 cells. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses predicted that their targets are enriched in cancer development-related pathways, such as MAPK signaling, microRNAs in cancer, apoptosis, PI3K-Akt signaling, cell cycle, Ras signaling, and Chronic myeloid leukemia. Eleven miRNAs were confirmed to have differential expression through qRT-PCR, with 6 upregulated (miR-140-3p, miR-551b-3p, miR-7-5p, miR-1290, miR-92a-3p, and miR-1246) and 5 downregulated (miR-183-5p, miR-26a-5p, miR-30c-5p, miR-205-5p, and miR-99b-3p). Among these, miR-1246 exhibited the highest expression level. HQ exposure resulted in a concentration-dependent increase in miR-1246 levels and decrease Cyclin G2 (CCNG2) levels in TK6 cells. Similarly, exosomes from HQ19 exhibited similar effects as HQ exposure. Dual luciferase reporter gene assays indicated that miR-1246 could band to CCNG2. After HQ exposure, exosomal miR-1246 induced cell cycle arrest at the S phase, elevating the expression of genes like pRb, E2F1, and Cyclin D1 associated with S phase checkpoint. However, silencing miR-1246 caused G2/M-phase arrest.
CONCLUSION
HQ-transformed cells' exosomal miR-1246 targets CCNG2, regulating TK6 cell cycle arrest, highlighting its potential as a biomarker for HQ-induced malignant transformation.
背景
对苯二酚（HQ）是苯的主要代谢产物，是一种已知的血液毒性致癌物。微小RNA 1246（miR-1246）作为一种癌基因，在包括白血病在内的肿瘤发生过程中调节靶基因。本研究调查了HQ转化（HQ19）细胞来源的外泌体miR-1246对受体TK6细胞间细胞通讯的影响。
方法
采用RNA测序鉴定HQ19细胞及其磷酸盐缓冲液对照细胞（PBS19）中差异表达的外泌体微小RNA，然后用qRT-PCR进行验证。研究了HQ转化细胞来源的外泌体miR-1246对受体TK6细胞周期分布的影响。
结果
RNA测序分析显示，与PBS19细胞相比，HQ19细胞中有34种外泌体微小RNA上调，158种微小RNA下调。基因本体论和京都基因与基因组百科全书分析预测，它们的靶标富集在癌症发展相关途径中，如MAPK信号通路、癌症中的微小RNA、细胞凋亡、PI3K-Akt信号通路、细胞周期、Ras信号通路和慢性粒细胞白血病。通过qRT-PCR确认有11种微小RNA存在差异表达，其中6种上调（miR-140-3p、miR-551b-3p、miR-7-（此处原文有误，应为miR-7-5p）、miR-1290、miR-92a-3p和miR-1246），5种下调（miR-183-5p、miR-26a-5p、miR-3c-5p（此处原文有误，应为miR-30c-5p）、miR-205-5p和miR-99b-3p）。其中，miR-1246表达水平最高。HQ暴露导致TK6细胞中miR-1246水平呈浓度依赖性增加，细胞周期蛋白G（此处原文有误，应为Cyclin G2）2（CCNG2）水平降低。同样，HQ19细胞的外泌体表现出与HQ暴露类似的效果。双荧光素酶报告基因检测表明，miR-1246可与CCNG2结合。HQ暴露后，外泌体miR-1246诱导细胞周期停滞在S期，提高了与S期检查点相关的pRb、E2F1和细胞周期蛋白D1等基因的表达。然而，沉默miR-1246会导致G2/M期停滞。
结论
HQ转化细胞的外泌体miR-1246靶向CCNG2，调节TK6细胞周期停滞，突出了其作为HQ诱导恶性转化生物标志物的潜力。
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