中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (9): 646-649.

• 论著 • 上一篇    下一篇

Notch1基因对裸鼠皮肤鳞状细胞癌移植瘤细胞增殖和凋亡的影响

刘冬1,夏永华1,李素娟2,3,张彩凤3,李敏4,付丹丹2,李占国3,田中伟5   

  1. 1. 新乡医学院第一附属医院皮肤性病科
    2. 新乡医学院第一附属医院皮肤科
    3.
    4. 河南省人民医院皮肤科
    5. 河南省新乡医学院第一附属医院皮肤科
  • 收稿日期:2010-11-08 修回日期:2011-05-04 出版日期:2011-09-15 发布日期:2011-08-31
  • 通讯作者: 田中伟 E-mail:zhonwt@163.com
  • 基金资助:

    河南省高校科技创新人才支持计划资助

Effects of Notch1 gene on the proliferation of and apoptosis in xenografted human cutaneous squamous cell carcinoma cells in nude mice

  • Received:2010-11-08 Revised:2011-05-04 Online:2011-09-15 Published:2011-08-31

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摘要：

目的 探讨Notch1基因在人皮肤鳞状细胞癌SCL-1细胞裸鼠移植瘤中的作用。方法 于裸鼠腋下接种0.2 ml 1 × 108/ml的SCL-1细胞悬液，建立人皮肤鳞状细胞癌裸鼠移植瘤模型，将其分为三组：未处理组（接种PBS悬浮的未转染细胞），空载体转染组（接种空载体转染的细胞）和Notch1转染组（接种Notch1转染的细胞）。连续2周观察转染荷瘤裸鼠的生长情况，采用TdT介导的dUTP缺口末端标记技术（TUNEL）研究肿瘤组织中的细胞凋亡，利用RT-PCR和Western印迹研究肿瘤组织中Notch1、bcl-2和bax mRNA和蛋白的表达。结果 Notch1转染组中肿瘤的生长明显受到抑制，其肿瘤的重量为（0.574 ± 0.219） g，显著低于未处理组（2.642 ± 0.404） g和空载体转染组（2.606 ± 0.512） g（F = 26.642，P 值均 < 0.01）。TUNEL结果表明，Notch1转染组每500个细胞中凋亡的细胞数为87 ± 9，明显高于未处理组（8 ± 2）和空载体转染组（10 ± 3）中细胞凋亡的数目（P < 0.05）；此外，RT-PCR和Western印迹结果表明Notch1转染组中裸鼠肿瘤组织Notch1和bax mRNA和蛋白表达均显著上升，而bcl-2的表达显著下调，三组间比较差异具有统计学意义（P值均 ＜ 0.05）。结论 Notch1基因能抑制人皮肤鳞状细胞癌细胞SCL-1裸鼠皮下移植瘤的生长，诱导SCL-1细胞凋亡，后者可能与bax表达的上升和bcl-2表达的下调相关。

关键词: 细胞凋亡

Abstract:

Objective To investigate the role of Notch1 gene in xenografted human cutaneous squamous cell （SCL-1） carcinoma. Methods Fifteen nude mice were divided into three groups, including untreated group （inoculated with SCL-1 cells treated with phosphate buffered saline）, empty vector group （inoculated with SCL-1 cells transfected with empty vector） and Notch1 group （inoculated with SCL-1 cells transfected with Notch1 expression vector）. All the mice were inoculated with SCL-1 cells （1 × 108/ml） of 0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay for the detection of cell apoptosis, reverse-transcription （RT）-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. Results The proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group （0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F = 26.642, P < 0.01）. TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was （87 ± 9） in Notch1 group, evidently higher than that in the untreated group （8 ± 2） and empty vector group （10 ± 3） （F = 194.266, P < 0.05）. Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch1 group, with significant difference among the three groups （all P < 0.05）. Conclusions Notch1 gene can inhibit the growth of xenografted human cutaneous squamous cell （SCL-1） carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

Key words: cell apoptosis