抗人IgM抗体对喉鳞癌Hep-2细胞裸鼠移植瘤血管生成的影响和机制研究

Effect of anti-human IgM antibody on angiogenesis in xenograft of Hep-2 human laryngeal squamous cell carcinoma line among nude mice

目的探讨抗人IgM抗体对人喉鳞癌Hep-2细胞裸鼠移植瘤血管生成的影响及其可能机制。方法先行制备Hep一2细胞裸鼠移植瘤,随机分为实验组(IEM组)、IgG对照组和模型对照组,每组5只。处理结束后,分别取移植瘤组织,免疫组化法检测各组移植瘤组织中CD31标记的微血管密度(MVD)及血管内皮生长因子(VEGF)、CD40蛋白表达,并行图像分析,比较分析各组间表达差异。结果实验组肿瘤组织MVD为(11.4±0.71),显著低于lgG对照组(13.8±1.57)、PBS对照组(16.28±1.85),Jp均<0.05;实验组VEGF平均光密度值(0.346±0.038),显著低于IgG对照组(0.367±0.041)和PBS对照组(0.370±0.023),P均<0.05;实验组CD40平均光密度值(0.172±0.016),显著低于IgG对照组(0.230±0.017)和PBS对照组(0.232±0.072),P均<0.05,各组瘤组织MVD与VEGF及CD40表达均成正相关(P均<0.05)。结论抗人IgM抗体可以抑制人喉鳞癌Hep-2细胞裸鼠移植瘤血管生成,其机制可能与抑制VEGF和CD40蛋白表达有关。

Objective To investigate the effect of anti-human IgM antibody on angiogenesis in transplanted tumor with cells of Hep-2 human laryngeal squamous cell carcinoma among nude mice and to explore its potential mechanism. Method Model of transplanted tumor was prepared among 15 nude mice with Hep-2 cells at first. Then, modeling animals were randomly divided into 5 groups, i.e. experimental group （EG）, IgG control group （ICG） and model group （MG）, with 5 mice in each group and treated with anti-human IgM antibody, IgG and PBS respectively. By the end of treating procedure, tissue samples were taken from transplanted tumor of each group and immunohistochemical assaying was made to determine microvessel density （MVD） labeled by CD31, and expressive activities of vascular endothelial growth factor （VEGF） as well as CD40 protein in tumor tissues, supplemented by image analysis to measure averaged optical density （OD） values of VEGF and CD40 protein, with a comparative analysis on their value differences among various groups. Result The value of MVD was （11.4＆#177;0.71） in EG, significantly lower than that in ICG （13.8＆#177;1.57） and MG （16.28＆#177;1.85） respectively （P＆lt;0.05）. Averaged OD of VEGF in EG was （0.346＆#177;0.038）, obviously lower than that in ICG （0.367＆#177;0.041） and MG （0.370＆#177;0.023） respectively （P＆lt;0.05）. The expressive intensity of CD40 in EG was （0.172＆#177;0.016）, also markedly lower than that in ICG （0.230＆#177;0.017） and MG （0.232＆#177;0.072） respectively （P＆lt;0.05）. There were positive correlations of MVD with the expressive activity either VEGF or CD40 among all these groups （P＆lt;0.05）. Conclusion Anti-human IgM antibody can inhibit angiogenesis in Hep-2 cell xenografts of human laryngeal squamous cell carcinoma among nude mice and its mechanism may be related to the inhibition of VEGF and CD40 expressive activities.