表达CRT/E7融合蛋白的人乳头瘤病毒DNA疫苗对肿瘤生长和血管生成抑制作用的研究HPV DNA Vaccines Expressing Recombinant CRT/ HPV6bE7 Fusion Protein Inhibit Tumor Growth and Angiogenic Activity
徐妍;程浩;赵可佳;朱可建;张行;
摘要(Abstract)：
研究HPV6b E7/CRT DNA疫苗免疫保护,清除已有感染和相关肿瘤细胞及其血管生成抑制作用,分析CRT抗血管生成的功能片段,为筛选高效的HPV疫苗提供实验依据。用重组质粒pcDNA3.1(+)-GFP-CRT120/HPV6bE7、pcDNA3.1(+)-GFP-HPV6bE7、pcDNA3.1(+)-GFP-CRT120、pcDNA3.1(+)-GFP-CRT180/HPV6bE7、pcDNA3.1(+)-GFP-CRT180通过肌内注射途径免疫C57BL/6小鼠。Matrigel法进行抗血管活性检测;B16/HPV6bE7细胞接种于C57BL/6雌性小鼠建立荷瘤模型,观察各组DNA疫苗对HPV6bE7基因的荷瘤组织的出瘤时间和肿瘤大小的影响。结果显示:重组DNA疫苗pcDNA3.1-CRT180/HPV6bE7和pcDNA3.1-CRT180在动物体内能对bFGF诱导的新生血管的生成有明显的抑制作用;CRT180/HPV6bE7和CRT180能显著抑制荷瘤的大小且CRT180/HPV6bE7免疫组较其他组能明显延缓荷瘤的形成时间、生长速率以及肿瘤重量。CRT180/HPV6bE7免疫组较其他组能诱导更强的血管抑制作用和部分抑制肿瘤生长,推测抑制血管的功能片段存在于CRT 120180 aa片段上。
关键词(KeyWords)： 尖锐湿疣;人乳头瘤病毒;钙网蛋白;重组DNA疫苗;血管抑制;抗肿瘤
基金项目(Foundation): 国家自然科学基金(30271190)
作者(Authors): 徐妍;程浩;赵可佳;朱可建;张行;
参考文献(References)：
- [1]Cheng W F,Hung C F,Chai C Y,et al.Tumor-specificimmunity and antiangiogenesis generated by a DNA vac-cine encoding calreticulin linked to a tumor antigen[J].JClin Invest,2001,108:669-678.
- [2]Ishiji T.Molecular mechanism of carcinogenesis by hu-man papillomavirus-16[J].J Dermatol,2000,27:73-86.
- [3]Peng S,Ji H,Trimble C,et al.Development of a DNAvaccine targeting human papillomavirus type 16 oncopro-tein E6[J].J Virol,2004,78:8468-8476.
- [4]Pike S E,Yao L,Jones K D,et al.Vasostatin,a calret-iculin fragment,inhibits angiogenesis and suppressestumor growth[J].J Exp Med,1998,188:2349-2356.
- [5]Pike S E,Yao L,Setsuda J,et al.Calreticulin and cal-reticulin fragments are endothelial cell inhibitors thatsuppress tumor growth[J].Blood,1999,94:2461-2468.
- [6]Yao L,Pike S E,Tosato G,et al.Laminin binding to thecalreticulin fragment vasostatin regulates endothelial cellfunction[J].J Leukoc Biol,2002,711:47-53.
- [7]Angiolillo A L,Sgadari C,Taub D D,et al.Human in-terferon-inducible protein 10 is a potent inhibitor of an-giogenesisin vivo[J].J Exp Med,1995,182:155-162.
- [8]Cheng W F,et al.Vascularity index as a novel parameterfor thein vivoassessment of angiogenesis in patientswith cervical carcinoma[J].Cancer,1999,85:651-657.
- [9]赵可佳,程浩,陈民利,等.表达人乳头瘤病毒6b和11型E6/E7基因的小鼠肿瘤细胞株的建立[J].中华皮肤科杂志,2004,37:91-93.
- [10]Nash P D,Opas M,Michalak M.Calreticulin:not justanother calcium-binding protein[J].Mol Cell Biochem,1994,135:71-78.