目的探讨新疆地区感染HPV16变异体谱系分布及E6、E7、LCR基因突变研究。方法对HPV16阳性的宫颈癌及癌前病变患者，提取基因组DNA，利用PCR扩增HPV16 DNA E6、E7基因及LCR区核苷酸片段，正反向测序。与HPV16基因序列分析比对，确定HPV16...目的探讨新疆地区感染HPV16变异体谱系分布及E6、E7、LCR基因突变研究。方法对HPV16阳性的宫颈癌及癌前病变患者，提取基因组DNA，利用PCR扩增HPV16 DNA E6、E7基因及LCR区核苷酸片段，正反向测序。与HPV16基因序列分析比对，确定HPV16谱系分布，分析核苷酸突变位点。结果E6基因突变率为80．00％，主要突变位点为T350G（59．78％）和T178G（18．48％）；E7突变率为54．78％，主要突变位点为A647G（33．33％）和T846C（26．98％）；LCR突变率为23．48％，主要突变位点为C24T（74．07％）和C13T（18．52％）。新疆维吾尔族妇女与汉族妇女之间比较，维吾尔族妇女T350G突变率（72．00％）显著高于汉族（45．24％），汉族妇女A647G（33．33％）、T846C（31．25％）、C24T（78．95％）三个突变率均显著高于维吾尔族（依次为20．00％、13．33％、62．50％），差异具有统计学意义。新疆汉族妇女中感染的HPV16主要为亚洲型，新疆维吾尔族妇女感染的HPV16主要为欧洲型。结论T350G突变可能是新疆维吾尔族妇女宫颈癌高发的原因之一，新疆汉族妇女中感染的HPV16型主要为亚洲型，新疆维吾尔族妇女感染的HPV16型主要为欧洲型。展开更多
Using human papillomavirus type 16 （HPV16） E7 as an antigen and Heat Shock Protein 70 as adjuvant, we constructed a DNA vaccine by linking HSP70 gene to E7^C91G; gene. Mice, after being immunized with E7^C91G;-HSP70...Using human papillomavirus type 16 （HPV16） E7 as an antigen and Heat Shock Protein 70 as adjuvant, we constructed a DNA vaccine by linking HSP70 gene to E7^C91G; gene. Mice, after being immunized with E7^C91G;-HSP70, E7^C91G/HSP70, E7^C91G, and wild E7 DNA vaccines respectively, produced E7 specific CD8^＋ T-cell precursor frequencies of 280.33±2.52, 144.34±4.04, 164.34±5.13 and 82.33±3.51 respectively within every 1 × 10^5 mouse splenocytes. This proves that E7^C91G-HSP70 fusion vaccine can significantly enhance the E7 specific cellular immunity within the mice body（p〈0. 01）. After being immunized with E7^C91G-HSP70 fusion vaccine, tumor-bearing mice of the group being treated have significantly longer latency and survival periods, comparing with other three categories of E7 vaccines. Experiment shows that this vaccine has a significant effect on enhancing E7 positive tumor-treatment within mice body. After being immunized with E7^C91G-HSP70 vaccine, there were no pathological changes found in livers, kidneys and spleens of the mice, which proves that the vaccine is quite safe. After all, E7^C91G-HSP70 fusion vaccine has a much stronger tumor- treatment effect than thai of wild type E7 DNA vaccine.展开更多
文摘目的探讨新疆地区感染HPV16变异体谱系分布及E6、E7、LCR基因突变研究。方法对HPV16阳性的宫颈癌及癌前病变患者，提取基因组DNA，利用PCR扩增HPV16 DNA E6、E7基因及LCR区核苷酸片段，正反向测序。与HPV16基因序列分析比对，确定HPV16谱系分布，分析核苷酸突变位点。结果E6基因突变率为80．00％，主要突变位点为T350G（59．78％）和T178G（18．48％）；E7突变率为54．78％，主要突变位点为A647G（33．33％）和T846C（26．98％）；LCR突变率为23．48％，主要突变位点为C24T（74．07％）和C13T（18．52％）。新疆维吾尔族妇女与汉族妇女之间比较，维吾尔族妇女T350G突变率（72．00％）显著高于汉族（45．24％），汉族妇女A647G（33．33％）、T846C（31．25％）、C24T（78．95％）三个突变率均显著高于维吾尔族（依次为20．00％、13．33％、62．50％），差异具有统计学意义。新疆汉族妇女中感染的HPV16主要为亚洲型，新疆维吾尔族妇女感染的HPV16主要为欧洲型。结论T350G突变可能是新疆维吾尔族妇女宫颈癌高发的原因之一，新疆汉族妇女中感染的HPV16型主要为亚洲型，新疆维吾尔族妇女感染的HPV16型主要为欧洲型。
基金Supported by the National Natural Science Foundation of China(30171042)
文摘Using human papillomavirus type 16 （HPV16） E7 as an antigen and Heat Shock Protein 70 as adjuvant, we constructed a DNA vaccine by linking HSP70 gene to E7^C91G; gene. Mice, after being immunized with E7^C91G;-HSP70, E7^C91G/HSP70, E7^C91G, and wild E7 DNA vaccines respectively, produced E7 specific CD8^＋ T-cell precursor frequencies of 280.33±2.52, 144.34±4.04, 164.34±5.13 and 82.33±3.51 respectively within every 1 × 10^5 mouse splenocytes. This proves that E7^C91G-HSP70 fusion vaccine can significantly enhance the E7 specific cellular immunity within the mice body（p〈0. 01）. After being immunized with E7^C91G-HSP70 fusion vaccine, tumor-bearing mice of the group being treated have significantly longer latency and survival periods, comparing with other three categories of E7 vaccines. Experiment shows that this vaccine has a significant effect on enhancing E7 positive tumor-treatment within mice body. After being immunized with E7^C91G-HSP70 vaccine, there were no pathological changes found in livers, kidneys and spleens of the mice, which proves that the vaccine is quite safe. After all, E7^C91G-HSP70 fusion vaccine has a much stronger tumor- treatment effect than thai of wild type E7 DNA vaccine.
