Yan Qin, Cheung Ying-Kit, Cheng Samuel Chak-Sum, Wang Xian-Hua, Shi Meng, Hu Mei-Haob, Yong Xie
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, PR China.
Gynecol Oncol. 2007 Jan;104(1):199-206. doi: 10.1016/j.ygyno.2006.07.044. Epub 2006 Oct 16.
Cervical cancer is found highly associated with human papillomaviruses type 16 (HPV16). HPV16 E6 and E7 oncogenes are important transforming genes which have become the main focus of anti-cervical cancer therapy. In this study, a recombinant DNA vaccine candidate, termed HPV16-DNA-E6E7, constructed with HPV16 E7 and E6 genes was generated and used to against HPV16-induced tumors.
We inserted an E7 DNA fragment into E6 gene to produce a recombinant gene (E6E7-DNA). The E6E7-DNA gene was inserted into a mammalian expression vector, pcDNA 3.1+, to construct the DNA vaccine candidate. Animals (C57BL/6 mice) were immunized with the vaccine candidate with various concentrations (50 microg, 100 microg or 200 microg, respectively), and cytotoxicity measurement and tumor protection assay were carried out to examine the immunological effects of the vaccine candidate.
Immunization of with HPV16-E6E7-DNA induced HPV16-specific immune response and also conveyed protection against TC-1 induced tumor in vivo. A survival rate (90%) after 45 days of tumor challenge was observed. The animals injected with a higher dosage of the vaccine (200 microg) exhibited prolonged survival duration of more than 55 days. No transforming activity of the vaccine candidate was detected, as determined by focus formation and degradation of endogenous p53.
Our results demonstrated that the HPV16-E6E7-DNA compound might become a candidate for HPV16 precautionary and immunotherapy.
宫颈癌被发现与16型人乳头瘤病毒(HPV16)高度相关。HPV16 E6和E7癌基因是重要的转化基因,已成为抗宫颈癌治疗的主要焦点。在本研究中,构建了一种用HPV16 E7和E6基因构建的重组DNA疫苗候选物,称为HPV16-DNA-E6E7,并用于对抗HPV16诱导的肿瘤。
我们将一个E7 DNA片段插入E6基因以产生一个重组基因(E6E7-DNA)。将E6E7-DNA基因插入哺乳动物表达载体pcDNA 3.1+中,构建DNA疫苗候选物。用不同浓度(分别为50微克、100微克或200微克)的疫苗候选物免疫动物(C57BL/6小鼠),并进行细胞毒性测定和肿瘤保护试验,以检测疫苗候选物的免疫效果。
用HPV16-E6E7-DNA免疫可诱导HPV16特异性免疫反应,并在体内对TC-1诱导的肿瘤产生保护作用。在肿瘤攻击45天后观察到存活率为90%。注射较高剂量疫苗(200微克)的动物存活时间延长超过55天。通过焦点形成和内源性p53降解测定,未检测到疫苗候选物的转化活性。
我们的结果表明,HPV16-E6E7-DNA复合物可能成为HPV16预防和免疫治疗的候选物。