中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (12): 843-846.

• 论著 • 上一篇    下一篇

光诱导纳米二氧化钛抑制人表皮鳞状细胞癌细胞株A431的实验研究

覃静净1,2,曾维惠3,高建武4,徐磊5,周莹6,耿松梅7   

  1. 1. 西安交通大学医学院第二附属医院皮肤科
    2. 西安医学院附属医院
    3.
    4. 陕西省人民医院皮肤科
    5. 成都市第二人民医院皮肤科
    6. 杭州浙江大学校医院华家池分院
    7. 西安交通大学第二附属医院皮肤科
  • 收稿日期:2011-12-13 修回日期:2012-08-14 出版日期:2012-12-15 发布日期:2012-11-30
  • 通讯作者: 曾维惠 E-mail:zengwh88@126.com
  • 基金资助:

    陕西省科技计划项目

Photo-induced inhibitory effect of titanium dioxide nanoparticles on a human epidermal squamous cell carcinoma cell line A431

  • Received:2011-12-13 Revised:2012-08-14 Online:2012-12-15 Published:2012-11-30

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摘要：

目的 探讨纳米二氧化钛（TiO2）颗粒光催化活性对人表皮鳞状细胞癌细胞株A431的生长抑制效应及机制。方法 单纯紫外线（主要波长为253.7 nm，功率30 W，光距30 cm，照射时间15 min）、不同浓度（100、200、300、400、500、600 mg/L）纳米TiO2及纳米TiO2 + 紫外线作用于A431细胞，采用噻唑蓝（MTT）法检测上述因素对A431细胞生长抑制率的影响；膜联蛋白Ⅴ-异硫氰酸荧光素/碘化丙啶（Annexin Ⅴ-FITC/PI）双染法检测A431细胞凋亡率；罗丹明123（Rho123）染色法检测A431细胞线粒体跨膜电位的变化。采用SPSS13.0统计软件进行t检验和方差分析，组间比较采用SNK检验。结果 实验干预24、48、72 h后，100、200、300、400、500、600 mg/L纳米TiO2 + 紫外线组A431细胞的生长抑制率增高，且呈浓度依赖效应，3个时间点不同浓度组比较，F值分别为21.54、77.56、20.27，P值均 ＜ 0.05（n = 6），SNK检验示各组间差异均有统计学意义；而不同浓度单纯纳米TiO2组细胞生长抑制率在3个时间点均无明显增高，差异均无统计学意义（P值均 ＞ 0.05）。纳米TiO2 联合紫外线照射可诱导A431细胞发生凋亡，并降低A431细胞线粒体跨膜电位，100、200、400 mg/L纳米TiO2 + 紫外线组凋亡率分别为8.86% ± 0.22%、11.72% ± 0.29%、31.24% ± 0.78%，空白对照组为2.69% ± 0.28%，经方差分析，差异有统计学意义（F = 256.61，P ＜ 0.05，n = 3）；以上3个浓度组线粒体跨膜电位总荧光强度值分别为758.48 ± 15.42、676.60 ± 14.35、557.71 ± 13.12，空白对照组为2943.65 ± 70.26，经方差分析，差异有统计学意义（F = 208.57，P ＜ 0.05，n = 3），SNK检验示各组间差异均有统计学意义。结论 纳米TiO2 + 紫外线对A431细胞有生长抑制作用，并能诱导细胞凋亡，线粒体跨膜电位下降可能是其诱导细胞凋亡的机制之一；单纯纳米TiO2对A431细胞的生长无抑制作用。

关键词: 细胞系，肿瘤

Abstract:

Objective To evaluate the inhibitory effect of photocatalytic titanium dioxide （TiO2）on the growth of a human epidermal squamous cell carcinoma cell line A431 and its mechanism. Methods Cultured A431 cells were classified into various groups to remain untreated （blank control group）, be treated with different concentrations （100, 200, 300, 400, 500, 600 mg/L） of TiO2 nanoparticles alone or in combination with ultraviolet（UV, main wavelength 253.7 nm, power 30 W, distance 30 cm, exposure duration 15 min） irradiation. After additional culture for different durations, methyl thiazolyl tetrazolium （MTT） assay was performed to evaluate cell growth, annexin V-fluorescein isothiocyanate/propidium iodide （PI） double staining to observe cell apoptosis, and Rho123 staining to determine mitochondrial transmembrane potential. Statistical analysis was carried out using SPSS 13.0 software. Analysis of variance （AOV）, t test and Student-Newman- Keuls （SNK） test were performed to assess the differences in these parameters between these groups. Results The growth of A431 cells was inhibited by pretreatment with TiO2 nanoparticles followed by UV irradiation, and the inhibitory effect was enhanced as the dose of TiO2 nanoparticles increased. As AOV and SNK test showed, there were significant differences in the growth inhibition rate among A431 cells treated with different concentrations of TiO2 nanoparticles at the three time points （24, 48 and 72 hours） after UV irradiation （n = 6, F = 21.54, 77.56, 20.27, respectively, all P < 0.05）. No statistical inhibition was observed in the growth of A431 cells treated with TiO2 nanoparticles alone compared with untreated A431 cells （all P > 0.05）. Photocatalytic TiO2 nanoparticles also induced the apoptosis but decreased the mitochondrial transmembrane potential in A431 cells. In detail, the apoptosis rate was 8.86% ± 0.22%, 11.72% ± 0.29% and 31.24% ± 0.78% in A431 cells treated with TiO2 nanoparticles of 100, 200, 400 mg/L followed by UV irradiation, respectively, compared to 2.69% ± 0.28% in the blank control group （n = 3, F = 256.61, P < 0.05）. Decreased mitochondrial transmembrane potential （expressed as total fluorescence intensity） was observed in A431 cells treated with TiO2 nanoparticles of 100, 200, 400 mg/L followed by UV irradiation compared with blank control group （758.48 ± 15.42, 676.60 ± 14.35, 557.71 ± 13.12 vs. 2943.65 ± 70.26, F = 208.57, P < 0.05, n = 3）, and SNK test also revealed statistical differences between these groups. Conclusions TiO2 nanoparticles combined with UV can inhibit the growth of but induce the apoptosis in A431 cells, which may be associated with the reduction in mitochondrial transmembrane potential in A431 cells, while TiO2 nanoparticles alone show no inhibitory effect on the growth of A431 cells.

中图分类号: 

  • R75