[紫外线照射对低氧条件下急性早幼粒细胞白血病细胞增殖的影响及相关机制]
[Effect of ultraviolet irradiation on the proliferation of acute promyelocytic leukemia cells under hypoxic conditions and related mechanisms].
作者信息
Wang Yi-Lin, Wang Ling-Zhen, Sun Jian-Dong, Li Xue-Rong, Wang Zhi, Sun Li-Rong
机构信息
Department of Pediatrics, Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, China.
出版信息
Zhongguo Dang Dai Er Ke Za Zhi. 2019 May;21(5):491-496. doi: 10.7499/j.issn.1008-8830.2019.05.018.
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/m, 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.
目的
研究280 nm发光二极管（LED）紫外线照射对缺氧条件下急性早幼粒细胞白血病（APL）HL-60细胞增殖的影响及其相关机制。
方法
选取对数生长期的HL-60细胞，分为对照组、缺氧组、紫外线组和缺氧+紫外线组。缺氧组细胞用氯化钴处理（终浓度为150 μmol/L），紫外线组用能量强度为30 J/m的280 nm LED紫外线照射，缺氧+紫外线组先用氯化钴处理，再用280 nm LED紫外线照射。处理48小时后，将细胞置于倒置显微镜下观察细胞形态。采用CCK-8法检测细胞增殖抑制率。用Annexin V-FITC/PI双染流式细胞术评估细胞凋亡。采用定量实时聚合酶链反应（qRT-PCR）检测Bcl-2的mRNA表达。上述每个实验均独立重复3次。
结果
与对照组相比，各实验组细胞均出现皱缩、亮度降低、排列紊乱，且随着培养时间延长细胞数量减少。各组细胞增殖抑制率和细胞凋亡率差异有统计学意义（P<0.01），缺氧+紫外线组对细胞增殖的抑制作用和诱导细胞凋亡作用最强，其次为紫外线组和缺氧组。与对照组相比，其他三组Bcl-2的mRNA表达均逐渐降低，且缺氧+紫外线组降低幅度明显大于缺氧组和紫外线组（P<0.01）。
结论
缺氧和紫外线照射均可抑制HL-60细胞增殖并诱导细胞凋亡，且紫外线照射在缺氧条件下对增殖抑制和细胞凋亡的作用优于常氧条件，其机制可能与下调Bcl-2的mRNA表达有关。
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