亚砷酸钠对SH-SY5Y细胞周期的影响及MPST过表达的干预
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摘要
目的探讨亚砷酸钠(NaAsO_2)对神经细胞周期的影响及MPST过表达在其中的作用。方法以空白组(正常SH-SY5Y细胞,BC)、空载对照组(NC)、过表达组(过表达MPST,OP)细胞分别经NaAsO_2处理后,采用CCK-8法、结晶紫染色法、流式细胞术和Western blot法,分别检测细胞活力、细胞贴壁力、细胞周期及p53、CDC25A、CyclinA、CDK2等蛋白表达水平。结果经NaAsO_2处理48 h后,NC组细胞存活率、细胞贴壁率明显降低,而过表达MPST细胞上述变化明显得到改善(P<0.01)。同时,NaAsO_2明显上调BC、NC组细胞S期比例和p53蛋白表达,而下调CDC25A、CyclinA及CDK2蛋白(P<0.01);然而,过表达MPST则明显拮抗上述效应(P<0.05,P<0.01)。结论 NaAsO_2通过诱导SH-SY5Y细胞S期阻滞,抑制细胞生长,而过表达MPST可拮抗NaAsO_2诱导的毒性效应。
Aim To investigate the effects of sodium arsenite(NaAsO_2) on the cell cycle and growth, and the intervention of MPST over-expression in the neural cells. Methods NaAsO_2 was used to treat SH-SY5Y neuroblastoma cells for 48 h from the blank control(BC),empty vector control(transfected with empty vector,NC) and over-expression group(lentiviral transfection with MPST,OP). The methods of CCK-8,crystal violet staining,flow cytometry and Western blot were used to examine cell viability,adherent rate,cell cycle and protein expression of p53,CDC25 A,CyclinA and CDK2. Results The cell viability and adherent rate significantly decreased after treatment with NaAsO_2 for 48 h,which was reversed in OP group( P < 0. 01).Meanwhile,NaAsO_2 also significantly increased the proportion of S phase cells and p53 protein expression,and down-regulated the protein levels of CDC25 A,Cyclin A and CDK2 in BC and NC groups( P < 0. 01),whereas the above changes of protein levels were significantly antagonized in OP group compared with NCgroup( P < 0. 05,P < 0. 01). Conclusions NaAsO_2 inhibits the cell growth by inducing S-phase arrest and over-expression of MPST could reverse the noxious effects caused by NaAsO_2 in SH-SY5Y cells.
Aim To investigate the effects of sodium arsenite(NaAsO_2) on the cell cycle and growth, and the intervention of MPST over-expression in the neural cells. Methods NaAsO_2 was used to treat SH-SY5Y neuroblastoma cells for 48 h from the blank control(BC),empty vector control(transfected with empty vector,NC) and over-expression group(lentiviral transfection with MPST,OP). The methods of CCK-8,crystal violet staining,flow cytometry and Western blot were used to examine cell viability,adherent rate,cell cycle and protein expression of p53,CDC25 A,CyclinA and CDK2. Results The cell viability and adherent rate significantly decreased after treatment with NaAsO_2 for 48 h,which was reversed in OP group( P < 0. 01).Meanwhile,NaAsO_2 also significantly increased the proportion of S phase cells and p53 protein expression,and down-regulated the protein levels of CDC25 A,Cyclin A and CDK2 in BC and NC groups( P < 0. 01),whereas the above changes of protein levels were significantly antagonized in OP group compared with NCgroup( P < 0. 05,P < 0. 01). Conclusions NaAsO_2 inhibits the cell growth by inducing S-phase arrest and over-expression of MPST could reverse the noxious effects caused by NaAsO_2 in SH-SY5Y cells.
引文
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[3] Li H,Chen G.In vivo reprogramming for CNS repair:regenerating neurons from endogenous glial cells[J].Neuron,2016,91(4):728-38.
[4] Qabazard B,Li L,Gruber J,et al.Hydrogen sulfide is an endogenous regulator of aging in Caenorhabditis elegans[J].Antioxid Redox Signal,2014,20(16):2621-30.
[5] 聂杨,王念,徐国强,等.亚砷酸钠对SH-SY5Y细胞生长和3MST表达的影响[J].中国药理学通报,2016,32(8):1182-3.[5] Nie Y,Wang N,Xu G Q,et al.Effect of sodium arsenite on cell growth and expression of 3MST in SH-SY5Y cells[J].Chin Pharmacol Bull,2016,32(8):1182-3.
[6] 王念,潘际刚,范海琼,等.亚砷酸钠对神经细胞活力及3MST蛋白表达的影响[J].现代预防医学,2017,44(11):2046-9.[6] Wang N,Pan J G,Fan H Q,et al.Effects of sodium arsenite on cell viability and expression of 3MST in neuronal cells[J].Mod Prev Med,2017,44(11):2046-9.
[7] Vimercati L,Gatti M F,Gagliardi T,et al.Environmental exposure to arsenic and chromium in an industrial area[J].Environ Sci Pollut Res Int,2017,24(12):11528-35.
[8] Ryu K H,Woo S Y,Lee M Y,et al.Morphological and biochemical changes induced by arsenic trioxide in neuroblastoma cell lines[J].Pediatr Hematol Oncol,2005,22(7):609-21.
[9] 胡亚男,赵巍,陈承志,等.亚砷酸钠和三氧化二砷对人正常肝细胞增殖与凋亡效应的影响[J].卫生研究,2014,43(2):203-9.[9] Hu Y N,Zhao W,Chen C Z,et al.Research on the sodium arsenite and arsenic trioxide induced proliferation and apoptosis effects on human hepatocyte[J].J Hyg Res,2014,43(2):203-9.
[10] Pozo-Molina G,Ponciano-Gómez A,Hernández-Zavala A,et al.Arsenic-induced S phase cell cycle lengthening is associated with ROS generation,p53 signaling and CDC25A expression[J].Chem Biol Interact,2015,238:170-9.
[11] Kokontis J M,Lin H P,Jiang S S,et al.Androgen suppresses the proliferation of androgen receptor-positive castration-resistant prostate cancer cells via inhibition of Cdk2,CyclinA,and Skp2[J].PLoS One,2014,9(10):e109170.
[12] 凌娜,孙庆岩,徐艳艳,等.硒化卡拉胶联合表阿霉素对肝癌HepG-2细胞增殖及细胞周期的影响[J].食品与药品,2014,16(2):81-4.[12] Ling N,Sun Q Y,Xu Y Y,et al.Effect of kappa-selenocarrageenan combined with epirubicin on proliferation and cell cycle of HepG-2 cells[J].Food Drug,2014,16(2):81-4.
[13] 董青川,程继,王志刚,等.miR-155通过p53/p21调控前列腺癌细胞周期[J].现代生物医学进展,2016,16(29):5640-3.[13] Dong Q C,Cheng J,Wang Z G,et al.Effect of miR-155 on the cell cycle of prostate cancer by p53/p21 pathways[J].Prog Mod Biomed,2016,16(29):5640-3.
[14] Chen Z,Zhang Z,Zhang D,et al.Hydrogen sulfide protects against TNF-α induced neuronal cell apoptosis through miR-485-5p/TRADD signaling[J].Biochem Biophys Res Commun,2016,478(3):1304-9.