Ang2-siRNA慢病毒干预裸鼠移植性恶性黑色素瘤的研究

目的研究血管生成素2小干扰RNA（Ang2-siRNA）对裸鼠移植性恶性黑色素瘤血管形成及其生长的影响。方法建立人恶性黑色素肿瘤裸鼠异种移植瘤模型。在体内使用pNL-EGFP—Ang2-siRNA慢病毒干扰裸鼠移植性恶性黑色素瘤Ang2基因的表达，观察统计各组肿瘤（空白组、空载组、实验组）生长情况。应用实时荧光定量RT—PCR检测肿瘤组织中Ang2基因的mRNA表达水平；CD34单克隆抗体作为血管内皮标志物，免疫组织化学法检测其表达，以评价肿瘤微血管密度。结果成功建立了恶性黑色素瘤裸鼠异种移植瘤模型，实验组肿瘤Ang2基因mRNA水平、微血管密度、肿瘤体积显著小于空白组和空载组（P〈0．01），空白组和空载组之间无统计学意义（P〉0．05）。结论pNL—EGFP—Ang2-siRNA慢病毒能沉默Ang2的表达，显著抑制恶性黑色素瘤血管的形成和肿瘤的生长，可能为临床恶性黑色素肿瘤的基因治疗开辟新的途径。

Ang2-siRNA Lentivirus Interference on Melanoma Xenograft in Nude Mice

Objective Research the influence of Ang2-siRNA on angiogenesis and growth of transplanted melanoma tumors in nude mice. Methods Human malignant melanoma （MM） trans- plantation tumor model was established in nude mice. Interference of Ang2 gene expression by pNL- EGFP-Ang2-siRNA lentivirus in transplanted MM was observed and recorded by comparison of tumor growth between the PBS control group, the Vector control group, and the experimental RNAi group. The expression levels of Ang2 gene in transplanted tumors was deteeted by Real-time RT- PCR. CD34, as one of the vascular endothelial markers, was detected by CD34 monoclonal anti- bodies to evaluate the microvessel deNsity of transplanted tumor using Immunohistochemical meth- od. Results The human MM transplanted tumor nude mice model was successfully established. The Ang2 gene mRNA level, microvassel deNsity, and tumor size in RNAi group were significantly de- ceased, compared to those in the PBS and Vector control groups （P 〈 0. 01 ） . There was no signif- icant difference between the PBS and Vector control groups （ P 〉 0. 05 ） . Conclusion PNL-EGFP- Ang2-siRNA lentivirus can reduce Ang2 gene expression, thereby inhibiting angiogenesis and the tumor growth, implying its potential application in clinical gene therapy of malignant melanomas.