探讨在体内和体外砷与Dicer表达的关系及Dicer在细胞增殖中的作用,体内实验选择云南某高污染砒霜厂工人作为砷暴露组,附近村庄无砷接触史的居民作为对照组,检测尿液中不同形态砷含量、外周血中Dicer mRNA表达水平.结果表明,与对照组相比,砷暴露组尿中外周血中Dicer mRNA的相对表达水平显著升高,Dicer mRNA表达水平与尿中无机砷、一甲基胂酸、二甲基胂酸呈正相关.体外实验以不同浓度(1.5,3,4.5μmol/L)的亚砷酸钠及4.5μmol/L的一甲基胂酸、二甲基胂酸、亚砷酸钠处理人上皮支气管细胞16HBE,检测Dicer mRNA和蛋白表达;敲低16HBE细胞内Dicer表达水平,检测细胞活力和增殖,结果表明,16HBE细胞中Dicer mRNA和Dicer蛋白表达水平在3,4.5μmol/L组均显著高于对照组,在一甲基胂酸组和二甲基砷酸组无变化.敲低Dicer抑制细胞活力和增殖,敲低Dicer和亚砷酸钠联合作用下,细胞增殖率下降更多.
Abstract
This study investigated the relationship between Dicer expression levels and urinary arsenic metabolites both in vivo and in vitro, as well as examined the role of Dicer in cell proliferation. For the epidemiological analysis, workers from a high arsenic-polluted factory in Yunnan Province were selected as the arsenic-exposed group, while residents from nearby villages without arsenic exposure history were recruited as controls. Urinary arsenic species (inorganic arsenic, monomethylarsonic acid [MMA], and dimethylarsinic acid [DMA]) were quantified, and Dicer mRNA expression levels in peripheral blood were measured. It was found that the relative Dicer mRNA expression in the arsenic-exposed group was significantly elevated compared to controls. Furthermore, Dicer mRNA levels were positively correlated with urinary inorganic arsenic, MMA, and DMA concentrations. For in vitro experiments, human bronchial epithelial cells (16HBE) were treated with sodium arsenite (1.5, 3, 4.5μmol/L) or 4.5μmol/L MMA, DMA, or sodium arsenite. Dicer mRNA and protein expression were analyzed by RT-qPCR and Western blot. Additionally, Dicer expression was knocked down in 16HBE cells using siRNA, and cell viability and proliferation were assessed via CCK-8 and EdU assays. It was observed that Dicer mRNA and protein levels in 3 and 4.5μmol/L sodium arsenite-treated cells were significantly upregulated compared to untreated controls, whereas no changes were detected in MMA- or DMA-treated groups. Knockdown of Dicer was shown to suppress cell viability and proliferation. Notably, sodium arsenite exposure combined with Dicer knockdown resulted in a more pronounced reduction in cell proliferation rates.
关键词
16HBE细胞 /
Dicer /
增殖 /
职业砷暴露
Key words
16HBE cells /
Dicer /
occupational arsenic exposure /
proliferation
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参考文献
[1] Nurchi V M, Djordjevic A B, Crisponi G, et al. Arsenic toxicity: Molecular targets and therapeutic agents[J]. Biomolecules, 2020,10(2): 235.
[2] Pershagen G. Lung cancer mortality among men living near an arsenic-emitting smelter[J]. American Journal of Epidemiology, 1985, 122:684-694.
[3] De Gregori I, Fuentes E, Rojas M, et al. Monitoring of copper, arsenic and antimony levels in agricultural soils impacted and non-impacted by mining activities, from three regions in Chile[J]. Journal of Environmental Monitoring, 2003,5:287-295.
[4] Sassano M, Seyyedsalehi Ms, Siea Ac, et al. Occupational arsenic exposure and digestive and head and neck cancers: A systematic review and meta-analysis[J]. Environmental Research, 2024,260: 119643.
[5] Folesani G, Galetti M, Petronini Pg, et al. Interaction between occupational and non-occupational arsenic exposure and tobacco smoke on lung cancerogenesis: A systematic review[J]. International Journal of Environmental Research and Public Health, 2023,20(5): 4167.
[6] Román M D, Niclis C, Aballay L R, et al. Do Exposure to arsenic, occupation and diet have synergistic effects on prostate cancer risk?[J]. Asian Pacific Journal of Cancer Prevention, 2018,19:1495-1501.
[7] Di Leva G, Garofalo M, Croce C M. MicroRNAs in cancer[J]. Annual Review of Pathology, 2014,9:287-314.
[8] Zapletal D, Kubicek K, Svoboda P, et al. Dicer structure and function: conserved and evolving features[J]. EMBO Reports, 2023,24(7): e57215.
[9] Vergani-Junior C A, Tonon-Da-Silva G, Inan M D, et al. DICER: structure, function, and regulation[J]. Biophysical Reviews, 2021, 13(6):1081-1090.
[10] Tseng C F, Chen L T, Wang H D, et al. Transcriptional suppression of Dicer by HOXB-AS3/EZH2 complex dictates sorafenib resistance and cancer stemness[J]. Cancer Science, 2022,113(5):1601-1612.
[11] Shan W, Sun C, Zhou B, et al. Role of Dicer as a prognostic predictor for survival in cancer patients: a systematic review with a meta-analysis[J]. Oncotarget, 2016,7(45):72672-72684.
[12] Zhao H, Jin X, Su H, et al. Down-regulation of Dicer expression in cervical cancer tissues[J]. Medical Oncology, 2014,31(5):937.
[13] Yu J, Li S, Shen S, et al. The transcript NR 134251.1of lncRNA APTR with an opposite function to all transcripts inhibits proliferation and induces apoptosis by regulating proliferation and apoptosis-related genes[J]. Human& Experimental Toxicology, 2023;42: 9603271221150247.
[14] Rager Je, Yosim A, Fry Rc. Prenatal exposure to arsenic and cadmium impacts infectious disease-related genes within the glucocorticoid receptor signal transduction pathway[J]. International Journal of Molecular Sciences, 2014,15(12):22374-22391.
[15] Shimoda Y, Kato K, Asami S, et al. Differences in apoptotic signaling and toxicity between dimethylmonothioarsinic acid (DMMTAV) and its active metabolite, dimethylarsinous acid (DMAIII), in HepaRG cells: Possibility of apoptosis cascade based on diversity of active metabolites of DMMTAV[J]. Journal of Trace Elements in Medicine and Biology, 2018,50:188-197.
[16] Suzuki T, Yamashita S, Ushijima T, et al. Genome-wide analysis of DNA methylation changes induced by gestational arsenic exposure in liver tumors[J]. Cancer Science, 2013,104(12):1575-1585.
[17] Sun M, Cheng H, Yu T, et al. Involvement of a AS3MT/c-Fos/ p53signaling axis in arsenic-induced tumor in human lung cells[J]. Environmental Toxicology, 2023,38(3):615-627.
[18] Wang X, Chen H, Wen Y, et al. Dicer affects cisplatin-mediated apoptosis in epithelial ovarian cancer cells[J]. Molecular Medicine Reports, 2018,18(5):4381-4387.
[19] Su Cm, Hsu Tw, Chen Ha, et al. Chaperone-mediated autophagy degrade Dicer to promote breast cancer metastasis[J]. Cellular Physiology, 2023,238(4):829-841.
[20] Faber C, Horst D, Hlubek F, et al. Overexpression of Dicer predicts poor survival in colorectal cancer[J]. European Journal of Cancer, 2011,47(9):1414-1419.
[21] Chiosea S, Jelezcova E, Chandran U, et al. Up-regulation of dicer, a component of the MicroRNA machinery, in prostate adenocarcinoma[J]. The American Journal of Pathology, 2006,169(5):1812-1820.
[22] Bakir M B, Salama M A, Refaat R, et al. Evaluating the therapeutic potential of one-carbon donors in nonalcoholic fatty liver disease[J]. European Journal of Pharmacology, 2019,847:72-82.
[23] Deshpande A A, Bhatia M, Laxman S, et al. Thiol trapping and metabolic redistribution of sulfur metabolites enable cells to overcome cysteine overload[J]. Microbial Cell, 2017,4(4):112-126.
[24] Kuo C C, Moon K A, Wang S L, et al. The association of arsenic metabolism with cancer, cardiovascular disease, and diabetes: A systematic review of the epidemiological evidence[J]. Environmental Health Perspectives, 2017,125(8):087001.
[25] Tan J, Sun M, Luo Q, et al. Arsenic exposure increased expression of HOTAIR and LincRNA-p21 in vivo and vitro[J]. Environmental Science and Pollution Research International, 2021,28(1):587-596.
[26] Wang M, Tan J, Jiang C, et al. Inorganic arsenic influences cell apoptosis by regulating the expression of MEG3 gene[J]. Environmental Geochemistry and Health, 2021,43(1):475-484.
[27] Mudhasani R, Zhu Z, Hutvagner G, et al. Loss of miRNA biogenesis induces p19Arf-p53signaling and senescence in primary cells[J]. The Journal of Cell Biology, 2008,181(7):1055-1063.
[28] Harris K S, Zhang Z, Mcmanus M T, et al. Dicer function is essential for lung epithelium morphogenesis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2006,103(7): 2208-2213.
[29] Roche B, Arcangioli B, Martienssen R. New roles for Dicer in the nucleolus and its relevance to cancer[J]. Cell Cycle, 2017,16(18): 1643-1653.
基金
云南省科技厅基础研究专项-面上项目(202401AT070177);国家自然科学基金(82160607);云南省创新团队(202405AS350016)
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