转染血小板第四因子氨基末端改构体cDNA抑制裸鼠实体肿瘤生长的研究

Transfer of cDNAs of platelet factor 4 and N-truncated peptide inhibits solid tumor growth in vivo

目的 研究血小板第四因子 (PF4 )氨基末端改构体cDNA (p17 70 )在实体肿瘤中抗血管新生基因治疗中的作用。方法 构建PF4及p17 70重组逆转录病毒载体 (pLXSN PF4和pLXSNp17 70 ) ,通过包装细胞系 (PA317 PF4和PA317p17 70 )产生的病毒上清转染人头颈部癌 (KB)细胞系 ,并建立KB细胞裸鼠移植瘤动物模型。体外实验观察MTT法检测转染的KB细胞增殖指数及重组逆转录病毒上清和重组分泌型PF4s对人脐静脉内皮细胞 (HUVEC)增殖的影响 ;体内实验 2 4只裸鼠 ,分KB PF4、KBP17 70、野生型KB组和空载体 4组 ,观察裸鼠移植瘤生长状态、死亡时间、及病理切片计数血管密度 ,并重复实验一次。结果 KB PF4s和野生型KB细胞增殖指数无显著差异 (P >0 0 5 ) ,PA317 PF4s病毒上清和KB PF4s细胞培养液可特异性抑制HUVEC增殖 (P <0 0 5 )。KB PF4和KBp17 70细胞在同源裸鼠体内生长速度减慢 (P <0 0 1,P <0 0 0 1)。与野生型KB组和空载体组相比较 (1 6 9g±0 35g ,1 94g± 0 2 5g) ,KB PF4组和KBp17 70组的瘤体平均重量明显减轻 (0 96g± 0 2 1g ,0 80g±0 34g) (P <0 0 5 )。实验荷瘤鼠生存时间延长 (P <0 0 5 ) ,移植瘤中新生血管密度明显减少 (P <0 0 0 1)。与KB PF4相比较 ,KBp17 70细胞在同源裸鼠体内生长速?

Objective Determine the anti tumor angiogenic effect of PF4 and p17 70 using virally mediated gene transfer Methods Full length PF4 cDNA and p17 70 cDNA were cloned into retroviral vector (pLXSN) KB was incubated with the supernatant of transfected PA317 PCR, RT PCR and Western blotting analysis was used to determine the integration and expression of foreign gene By MTT method the effects of the supernatants of PA317 PF4s and KB PF4s on the proliferation of human umbilical vein endothelial cells (HUVEC) were studied Tumorigenecity of KB PF4 and KBp17 70 cells was assayed with xenograft tumor growth in isogenous nude mice by examining the growth rate of xenograft, survival, and histochemistry of xenograft tumor Results Recombinant PF4 and p17 70 were able to inhibit selectively HUVEC proliferation Animal survival was significantly prolonged Furthermore, p17 70 significantly prolonged animal survival compared with PF4 group ( P <0 05) Conclusion Transduction of p17 70 inhibits solid tumor growth through an anti angiogensis mechanism Targeted anti angiogenesis, using retroviral mediated PF4 gene transfer, especially p17 70 represents a promising strategy for cancer gene therapy