Camk2b低表达对1,4-苯醌致K562细胞线粒体毒性的影响

doi:10.3969/j.issn.1004-616x.2019.04.001

Abstract

Abstract: OBJECTIVE:To investigate the effect from exposure to 1,4-benzoquinone (1,4-BQ) and of low expression of calcium/calmodulin dependent protein kinase 2 beta (Camk2b) on mitochondrial toxicity in K562 cells. METHODS:Expression of Camk2b mRNA in K562 cells was determined after 1,4-BQ exposure. Then,the appropriate concentrations of Camk2b inhibitor,KN93,was selected. The gene and protein expressions of Camk2b were verified by real-time PCR and Western blot,respectively,and the low expression of Camk2b was successfully constructed. After K562 cells were exposed to 0,10,and 20 μmol/L 1,4-BQ,the following assays were conducted:cell proliferation,reactive oxygen species,ATP production,mitochondrial membrane potential,calcium ion and apoptosis. The same assays were conducted in K562 cells and control cells which showed low expression of Camk2b. RESULTS:Expression of Camk2b mRNA in K562 cells was reduced after 1,4-BQ exposure. Compared with the control cells,significant changes of the followings were detected:reduction of the ATP production of Camk2b low expression cells,reduction of the mitochondrial membrane potentials,increase of the calcium ion concentrations and increase of the apoptotic rates (all P < 0.01). After 1,4-BQ exposure and compared with the same concentrations of 1,4-BQ-treated control cells,significant changes were observed:increase of the ROS level of Camk2b low expression cells,reduction of ATP production,decrease of mitochondrial membrane potentials,increased of the calcium concentrations and increase of the apoptotic rates (P < 0.05 or 0.01). CONCLUSION:After the K562 cells were exposed to 1,4-BQ,expression of Camk2b mRNA was reduced. Compared with the control cells,Camk2b low-expression cells showed more obvious mitochondrial damage after the 1,4-BQ exposure and Camk2b was apparently important for antioxidant damage to these cells.

Key words: camk2b, 1,4-benzoquinone, mitochondrial damage, K562 cells

CLC Number:

R135.1

ZHANG Hong, WANG Tong, WANG Kun, WANG Boshen, ZHANG Mengying, ZHOU Yanhua, PU Yuepu, ZHANG Juan. Exposure to 1,4-benzoquinone and low expression of Camk2b on mitochondrial toxicity in K562 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(4): 261-267.

References

[1] DOUGHERTY D, GARTE S, BARCHOWSKY A, et al. NQO1, MPO, CYP2E1, GSTT1 and GSTM1 polymorphisms and biological effects of benzene exposure:a literature review[J]. Toxicol Lett, 2008, 182(1/2/3):7-17.
[2] MCHALE C M, ZHANG L P, SMITH M T. Current understanding of the mechanism of benzene-induced leukemia in humans:implications for risk assessment[J]. Carcinogenesis, 2012, 33(2):240-252.
[3] CASALE T, SACCO C, RICCI S, et al. Workers exposed to low levels of benzene present in urban air:Assessment of peripheral blood count variations[J]. Chemosphere, 2016, 152:392-398.
[4] SNYDER R. Leukemia and benzene[J]. Int J Environ Res Public Health, 2012, 9(8):2875-2893.
[5] HUANG J S, ZHAO M D, LI X J, et al. The cytotoxic effect of the benzene metabolite hydroquinone is mediated by the modulation of MDR1 expression via the NF-κB signaling pathway[J]. Cell Physiol Biochem, 2015, 37(2):592-602.
[6] SMITH M T, ZHANG L P, MCHALE C M, et al. Benzene, the exposome and future investigations of leukemia etiology[J]. Chem Biol Interact, 2011, 192(1/2):155-159.
[7] 蒋小云, 孙蓉丽, 满招娣, 等. 1, 4-苯醌致K562细胞线粒体功能障碍与凋亡作用[J]. 癌变·畸变·突变, 2017, 29(2):129-133.
[8] WANG Q, GUO W J, HAO B X, et al. Mechanistic study of TRPM2-Ca(2+)-CAMK2-BECN1 signaling in oxidative stress-induced autophagy inhibition[J]. Autophagy, 2016, 12(8):1340-1354.
[9] LI C N, LI L, LIN B C, et al. Tris (1, 3-dichloro-2-propyl) phosphate induces toxicity by stimulating CaMK2 in PC12 cells[J]. Environ Toxicol, 2017, 32(6):1784-1791.
[10] 刘巍, 魏兴龙, 杨天瑶, 等. 线粒体钙超载激活线粒体凋亡途径在甲基汞致神经元凋亡中的作用[J]. 实用预防医学, 2018, 25(12):1409-1412.
[11] 曹萌. G6PD对苯暴露小鼠骨髓细胞信号通路影响的研究[D]. 南京:东南大学, 2017.
[12] WANG M G, LI W H, WANG X Y, et al. CaMKⅡ is involved in subcellular Ca²⁺ redistribution-induced endoplasmic reticulum stress leading to apoptosis in primary cultures of rat proximal tubular cells exposed to lead[J]. Oncotarget, 2017, 8(53):91162-91173.
[13] ZHONG P, QUAN D J, PENG J Y, et al. Role of CaMKⅡ in free fatty acid/hyperlipidemia-induced cardiac remodeling both in vitro and in vivo[J]. J Mol Cell Cardiol, 2017, 109:1-16.
[14] 刘子闲, 董芳, 张森, 等. 线粒体调控造血干细胞功能的研究进展[J]. 中国实验血液学杂志, 2019, 27(1):277-282.
[15] ARUN S, LIU L, DONMEZ G. Mitochondrial biology and neurological diseases[J]. Curr Neuropharmacol, 2016, 14(2):143-154.
[16] 杨婷婷, 江振洲, 张陆勇. 经由线粒体损伤诱发的药源性肝损伤研究进展[J]. 药学进展, 2014, 38(11):809-818.
[17] WANG L, WU Q, FAN Z J, et al. Platelet mitochondrial dysfunction and the correlation with human diseases[J]. Biochem Soc Trans, 2017, 45(6):1213-1223.
[18] XU S N, WANG T S, LI X, et al. SIRT2 activates G6PD to enhance NADPH production and promote leukaemia cell proliferation[J]. Sci Rep, 2016, 6:32734.
[19] PINTON P, GIORGI C, SIVIERO R, et al. Calcium and apoptosis:ER-mitochondria Ca²⁺ transfer in the control of apoptosis[J]. Oncogene, 2008, 27(50):6407-6418.
[20] LIU M J, WANG Z, JU Y, et al. Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca²⁺ homeostasis[J]. Cancer Chemother Pharmacol, 2005, 55(1):79-90.
[21] CIRCU M L, AW T Y. Reactive oxygen species, cellular redox systems, and apoptosis[J]. Free Radic Biol Med, 2010, 48(6):749-762.
[22] KUANG C Y, YU Y, WANG K, et al. Knockdown of transient receptor potential canonical-1 reduces the proliferation and migration of endothelial progenitor cells[J]. Stem Cells Dev, 2012, 21(3):487-496.
[23] MONACO S, RUSCIANO M R, MAIONE A S, et al. A novel crosstalk between calcium/calmodulin kinases Ⅱ and IV regulates cell proliferation in myeloid leukemia cells[J]. Cell Signal, 2015, 27(2):204-214.
[24] AN P, ZHU J Y, YANG Y, et al. KN-93, a specific inhibitor of CaMKⅡ inhibits human hepatic stellate cell proliferation in vitro[J]. World J Gastroenterol, 2007, 13(9):1445-1448.

Exposure to 1,4-benzoquinone and low expression of Camk2b on mitochondrial toxicity in K562 cells

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[3]	. [J]. , 2010, 22(2): 146-148.
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[6]	. Review; [J]. , 2000, 12(4): 227-228.