PDF(1922 KB)
紫外线在低氧条件下对急性早幼粒白血病细胞增殖的影响及机制研究
王艺霖, 王玲珍, 孙健栋, 李学荣, 王志, 孙立荣
中国当代儿科杂志 ›› 2019, Vol. 21 ›› Issue (5) : 491-496.
PDF(1922 KB)
PDF(1922 KB)
紫外线在低氧条件下对急性早幼粒白血病细胞增殖的影响及机制研究
Effect of ultraviolet irradiation on the proliferation of acute promyelocytic leukemia cells under hypoxic conditions and related mechanisms
目的 观察280 nm波长紫外发光二极管照射在低氧条件下对急性早幼粒白血病细胞（HL-60）增殖的影响，并探讨发生机制。方法 取对数生长期的HL-60细胞为研究对象，分为对照组、低氧组、紫外线组及低氧+紫外线组。低氧组给予氯化钴（终浓度150 μmol/L）处理，紫外线组用30 J/m2能量的280 nm波长紫外发光二极管照射，低氧+紫外线组在氯化钴处理基础上用30 J/m2能量的280 nm波长紫外发光二极管照射，48 h后收集细胞于倒置显微镜下观察细胞形态变化。采用CCK-8法检测细胞增殖抑制率；Annexin-FITC/PI双染流式细胞术检测细胞凋亡；实时荧光定量PCR检测Bcl-2基因mRNA的表达量。上述各实验均独立重复3次。结果 与对照组相比，各实验组细胞皱缩、透亮度降低、排列紊乱，随着培养时间延长，细胞数量减少。各组细胞增殖抑制率及细胞凋亡率比较差异有统计学意义（P < 0.01），其中低氧组+紫外线组细胞增殖抑制及诱导细胞凋亡作用最强，紫外线组次之，低氧组最弱。与对照组比较，低氧组、紫外线组及低氧+紫外线组Bcl-2 mRNA的表达逐渐下调，且低氧+紫外线组Bcl-2 mRNA表达量较低氧组和紫外线组下调更明显（P < 0.05）。结论 低氧和紫外线作用均可抑制HL-60细胞增殖，诱导细胞凋亡，紫外线在低氧条件下对细胞的增殖抑制及凋亡作用较常氧条件下更明显，其机制可能与Bcl-2基因表达下调有关。
Objective To study the effect of 280 nm-LED ultraviolet irradiation on the proliferation of acute promyelocytic leukemia (APL) HL-60 cells under hypoxic conditions and related mechanism. Methods HL-60 cells in the logarithmic growth phase were selected and divided into control, hypoxia, ultraviolet and hypoxia+ultraviolet groups. The cells in the hypoxia group were treated with cobalt chloride (with a final concentration of 150 μmol/L), those in the ultraviolet group were irradiated by 280 nm-LED ultraviolet with an energy intensity of 30 J/m2, and those in the hypoxia+ultraviolet group were treated with cobalt chloride and then irradiated by 280 nm-LED ultraviolet. After 48 hours of treatment, the cells were placed under an invert microscope to observe cell morphology. CCK-8 assay was used to measure the inhibition rate of cell proliferation. Annexin V-FITC/PI double staining flow cytometry was used to evaluate cell apoptosis. Quantitative real-time PCR was used to measure the mRNA expression of Bcl-2. Each experiment above was repeated three times independently. Results Compared with the control group, the experimental groups showed shrinkage, decreased brightness, and disordered arrangement of cells, and the number of cells decreased over the time of culture. There were significant differences in the inhibition rate of cell proliferation and cell apoptosis rate among the groups (P < 0.01), and the hypoxia+ultraviolet group showed the strongest inhibition of cell proliferation and induction of cell apoptosis, followed by the ultraviolet group and the hypoxia group. Compared with the control group, the other three groups had a gradual reduction in the mRNA expression of Bcl-2, and the hypoxia+ultraviolet group had a significantly greater reduction than the hypoxia and ultraviolet groups (P < 0.01). Conclusions Both hypoxia and ultraviolet irradiation can inhibit the proliferation of HL-60 cells and induce cell apoptosis, and ultraviolet irradiation has a better effect on proliferation inhibition and cell apoptosis under hypoxic conditions than under normoxic conditions, possibly by downregulating the mRNA expression of Bcl-2.
白血病 / 紫外线 / 低氧 / 细胞凋亡 / HL-60细胞
Leukemia / Ultraviolet / Hypoxia / Cell apoptosis / HL-60 cell
[1] Vellenga E, van Putten W, Ossenkoppele GJ, et al. Autologous peripheral blood stem cell transplantation for acute myeloid leukemia[J]. Blood, 2011, 118(23):6037-6042.
[2] Wetzel D, Mueller BU, Mansouri TB, et al. Delayed Haematological recovery after autologous stem cell transplantation is associated with favourable outcome in acute myeloid leukaemia[J]. Br J Haematol, 2015, 168(2):268-273.
[3] 曹易耕, 姜尔烈, 何祎, 等. 自体外周血造血干细胞移植治疗55例急性髓系白血病患者的疗效及预后分析[J]. 中华血液学杂志, 2016, 37(6):464-468.
[4] Azzouz D, Khan MA, Sweezey N, et al. Two-in-one:UV radiation simultaneously induces apoptosis and NETosis[J]. Cell Death Discov, 2018, 4:51.
[5] Harberts E, Fishelevich R, Liu J, et al. MyD88 mediates the decision to die by apoptosis or necroptosis after UV irradiation[J]. Innate Immun, 2014, 20(5):529-539.
[6] Salucci S, Burattini S, Battistelli M, et al. Ultraviolet B (UVB) irradiation-induced apoptosis in various cell lineages in vitro[J]. Int J Mol Sci, 2012, 14(1):532-546.
[7] Artiukhov VG, Zemchenkova OV, Basharina OV, et al. Apoptosis and necrosis of lymphocytes induced by UV-radiation in the presence of autological plasma[J]. Tsitologiia, 2014, 56(1):77-83.
[8] Song K, Mohseni M, Taghipour F. Application of ultraviolet light-emitting diodes (UV-LEDs) for water disinfection:a review[J]. Water Res, 2016, 94:341-349.
[9] Kemeny L, Csoma Z, Bagdi E, et al. Targeted phototherapy of plaque-type psoriasis using ultraviolet B-light-emitting diodes[J]. Br J Dermatol, 2010, 163(1):167-173.
[10] Hu Z, Wang F, Wu Z, et al. FOXO3a-dependent up-regulation of Mxi1-0 promotes hypoxia-induced apoptosis in endothelial cells[J]. Cell Signal, 2018, 51:233-242.
[11] Zhou J, Qian CY, Tong RQ, et al. Hypoxia induces apoptosis of mouse spermatocyte GC-2 cells through activation of autophagy[J]. Cell Biol Int, 2018. doi:10.1002/cbin.10971.[Epub ahead of print]
[12] Teti G, Focaroli S, Salvatore V, et al. The hypoxia-mimetic agent cobalt chloride differently affects human mesenchymal stem cells in their chondrogenic potential[J]. Stem Cells Int, 2018, 2018:3237253.
[13] 石旭. 低氧模拟物CoCl2对人急性髓系白血病细胞HL60中ROS及钙离子的调控及其意义[D]. 北京:中国协和医科大学, 2008.
[14] Xie D, Sun Y, Wang L, et al. Ultraviolet light-emitting diode irradiation-induced cell death in HL-60 human leukemia cells in vitro[J]. Mol Med Rep, 2016, 13(3):2506-2510.
[15] Reichrath J, Rass K. Ultraviolet damage, DNA repair and vitamin D in nonmelanoma skin cancer and in malignant melanoma:an update[J]. Adv Exp Med Biol, 2014, 810:208-233.
[16] Synowiec E, Hoser G, Wojcik K, et al. UV differentially induces oxidative stress, DNA damage and apoptosis in BCR-ABL1-positive cells sensitive and resistant to imatinib[J]. Int J Mol Sci, 2015, 16(8):18111-18128.
[17] de Pedro I, Alonso-Lecue P, Sanz-Gómez N, et al. Sublethal UV irradiation induces squamous differentiation via a p53-independent, DNA damage-mitosis checkpoint[J]. Cell Death Dis, 2018, 9(11):1094.
[18] Yogosawa S, Yoshida K. Tumor suppressive role for kinases phosphorylating p53 in DNA damage-induced apoptosis[J]. Cancer Sci, 2018, 109(11):3376-3382.
[19] Shokrollahi BM, Hasanzadeh H, Barati M, et al. Apoptosis/necrosis induction by ultraviolet, in ER positive and ER negative breast cancer cell lines[J]. Iran J Cancer Prev, 2015, 8(6):e4193.
[20] Kong L, Wang S, Wu X, et al. Paeoniflorin attenuates ultraviolet B-induced apoptosis in human keratinocytes by inhibiting the ROS-p38-p53 pathway[J]. Mol Med Rep, 2016, 13(4):3553-3558.
[21] Srinivas US, Tan BWQ, Vellayappan BA, et al. ROS and the DNA damage response in cancer[J]. Redox Biol, 2018. doi:10.1016/j.redox.2018.101084.[Epub ahead of print]
[22] Widel M, Krzywon A, Gajda K, et al. Induction of bystander effects by UVA, UVB, and UVC radiation in human fibroblasts and the implication of reactive oxygen species[J]. Free Radic Biol Med, 2014, 68:278-287.
[23] Ali D, Verma A, Mujtaba F, et al. UVB-induced apoptosis and DNA damaging potential of chrysene via reactive oxygen species in human keratinocytes[J]. Toxicol Lett, 2011, 204(2-3):199-207.
[24] Baharaghdam S, Yousefi M, Movasaghpour A, et al. Effects of hypoxia on biology of human leukemia T-cell line (MOLT-4 cells) co-cultured with bone marrow mesenchymal stem cells[J]. Avicenna J Med Biotechnol, 2018, 10(2):62-68.
[25] 张博, 翁勤洁, 陈中婷, 等. Q39在缺氧条件下诱导人白血病K562细胞凋亡[J]. 浙江大学学报(医学版), 2007, 36(3):261-266.
[26] Peng C, Rao W, Zhang L, et al. Mitofusin 2 ameliorates hypoxia-induced apoptosis via mitochondrial function and signaling pathways[J]. Int J Biochem Cell Biol, 2015, 69:29-40.
[27] Muñoz-Sánchez J, Chánez-Cárdenas ME. The use of cobalt chloride as a chemical hypoxia model[J]. J Appl Toxicol, 2018. doi:10.1002/jat.3749.[Epub ahead of print]
[28] Lee M, Kang H, Jang SW. CoCl2 induces PC12 cells apoptosis through p53 stability and regulating UNC5B[J]. Brain Res Bull, 2013, 96:19-27.
[29] Yang Z, Yang C, Xiao L, et al. Novel insights into the role of HSP90 in cytoprotection of H2S against chemical hypoxia-induced injury in H9c2 cardiac myocytes[J]. Int J Mol Med, 2011, 28(3):397-403.
[30] Dai ZJ, Gao J, Ma XB, et al. Up-regulation of hypoxia inducible factor-1alpha by cobalt chloride correlates with proliferation and apoptosis in PC-2 cells[J]. J Exp Clin Cancer Res, 2012, 31:28.
[31] Si H, Zhang Y, Song Y, et al. Overexpression of adrenomedullin protects mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the Akt/GSK3β and Bcl-2 signaling pathways[J]. Int J Mol Med, 2018, 41(6):3342-3352.
[32] Birkinshaw RW, Czabotar PE. The BCL-2 family of proteins and mitochondrial outer membrane permeabilisation[J]. Semin Cell Dev Biol, 2017, 72:152-162.
