Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi pr...Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi province; in addition variation of CTSEs was analyzed through single-strand conformation polymorphisms (SSCP). Results HPV16 CTSEs were detectable in 34 of 58 (57%) specimens and mutant rate was 41%(14/34) and the main mutations of chosen randomly variant CTSE (CTSEv) happened at YY1 binding sites in addition to glucocoticoid response elements (GRE). Conclusion CTSE in some specimens of Shaanxi province was obviously different from that in HPV16 wild type and variant CTSE might affect the transcriptional regulation of LCR on viral P97, which regulates over-expression of viral oncogenes in cervical carcinoma.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
Infection with high risk human papillomavirus is regarded as the major risk factor in the development of cervical cancer. In this study, HPV16 L1 eukaryotic expression plasmids pcDNA LI were constructed, which were...Infection with high risk human papillomavirus is regarded as the major risk factor in the development of cervical cancer. In this study, HPV16 L1 eukaryotic expression plasmids pcDNA LI were constructed, which were transfected into mammalian cells Cos 7. The expression of HPV16 L1 in transfected cells were identified by in situ hybridization, immunospot and immunocytochemistry. HPV16 L1 mRNA transcription and L1 protein expression were found in recombinant plasmid transfected cells. This expression system will provide us with plentiful resource for HPV16 L1 immunological study and will be helpful for the design of HPV16 prophylactic vaccine.展开更多
In order to provide the experimental basis for the further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16(HPV16),the eukaryotie expression vector pcDNA3.1(-)/E6 was used for the...In order to provide the experimental basis for the further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16(HPV16),the eukaryotie expression vector pcDNA3.1(-)/E6 was used for the study on the effect of E6 protein to influence the secretory activity of LPS-indueed THP-1-macrophages,and the reconstructed plasmid pcDNA3.1(-)/E6 was transfected into THP-1-maerophages.The expression of E6 gene was assayed in macrophage lysates by using Western blot analysis and the level of TNF-αor IL-1βwas examined by ELISA.All of data were analyzed by SPSS12.0.As demonstrated by Western blot analysis,the expression of E6 protein with a molecular weight of about 18 kDa by plasmid pcDNA3.1(-)/E6 in THP-1-macrophages could be detected.Howev- er,as demonstrated by ELISA assay,the level of TNF-αor IL-1βin lysates of THP-1-macrophages showed an obvious difference between the pcDNA3.1(-)/E6 group and the LPS control group or the pcDNA3.1(-)control group(P<0.01),but no significant difference existed between pcDNA3.1(-) control group and LPS control group(P>0.05).All these results illustrate that the transient over-ex- pression of HPV6 E6 protein reduces the production of TNF-αand IL-1βinduced by LPS in THP-1-mac- rophages.展开更多
目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性...目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性增强子(cell type specific enhancer,CTSE)。结果:在40例食管鳞癌中,HPV16的E6、E7基因和CTSE片段的检出率分别是37.5%(15/40)、42.5%(17/40)和40%(16/40),20例正常食管黏膜中E6、E7和CTSE的检出率分别为0%(0/20)、0%(0/20)和5%(1/20),两者均存在显著性差异(P<0.05)。CTSE片段分别与E6和E7基因有明显相关性(P<0.05)。食管鳞癌中E6、E7基因及CTSE的检出率在患者不同性别、年龄、肿瘤浸润程度、淋巴结转移和组织学分级肿瘤中差异均无显著性(P>0.05)。结论:E6和E7基因与CTSE片段共存于HPV16感染的食管鳞癌组织中,三者可能与食管鳞癌的发生和发展有关。展开更多
基金The study was supported by National Natural Science Foundation of China (No.39970035) .
文摘Objective To investigate the cell-type-specific enhancer (CTSE) in HPV16 and its variation in cervical carcinoma. Methods CTSEs were detected by polymerase chain reaction (PCR) in 58 cervical carcinoma from Shaanxi province; in addition variation of CTSEs was analyzed through single-strand conformation polymorphisms (SSCP). Results HPV16 CTSEs were detectable in 34 of 58 (57%) specimens and mutant rate was 41%(14/34) and the main mutations of chosen randomly variant CTSE (CTSEv) happened at YY1 binding sites in addition to glucocoticoid response elements (GRE). Conclusion CTSE in some specimens of Shaanxi province was obviously different from that in HPV16 wild type and variant CTSE might affect the transcriptional regulation of LCR on viral P97, which regulates over-expression of viral oncogenes in cervical carcinoma.
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.
文摘Infection with high risk human papillomavirus is regarded as the major risk factor in the development of cervical cancer. In this study, HPV16 L1 eukaryotic expression plasmids pcDNA LI were constructed, which were transfected into mammalian cells Cos 7. The expression of HPV16 L1 in transfected cells were identified by in situ hybridization, immunospot and immunocytochemistry. HPV16 L1 mRNA transcription and L1 protein expression were found in recombinant plasmid transfected cells. This expression system will provide us with plentiful resource for HPV16 L1 immunological study and will be helpful for the design of HPV16 prophylactic vaccine.
文摘In order to provide the experimental basis for the further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16(HPV16),the eukaryotie expression vector pcDNA3.1(-)/E6 was used for the study on the effect of E6 protein to influence the secretory activity of LPS-indueed THP-1-macrophages,and the reconstructed plasmid pcDNA3.1(-)/E6 was transfected into THP-1-maerophages.The expression of E6 gene was assayed in macrophage lysates by using Western blot analysis and the level of TNF-αor IL-1βwas examined by ELISA.All of data were analyzed by SPSS12.0.As demonstrated by Western blot analysis,the expression of E6 protein with a molecular weight of about 18 kDa by plasmid pcDNA3.1(-)/E6 in THP-1-macrophages could be detected.Howev- er,as demonstrated by ELISA assay,the level of TNF-αor IL-1βin lysates of THP-1-macrophages showed an obvious difference between the pcDNA3.1(-)/E6 group and the LPS control group or the pcDNA3.1(-)control group(P<0.01),but no significant difference existed between pcDNA3.1(-) control group and LPS control group(P>0.05).All these results illustrate that the transient over-ex- pression of HPV6 E6 protein reduces the production of TNF-αand IL-1βinduced by LPS in THP-1-mac- rophages.
文摘目的:检测食管鳞癌组织中人乳头瘤病毒(HPV)16E6、E7基因和细胞特异性增强子片段(CTSE)。方法:采用聚合酶链反应法(PCR)检测40例食管鳞癌和20例正常食管黏膜中HPV16E6、E7基因和病毒长控制区内(long control region,LCR)的细胞型特异性增强子(cell type specific enhancer,CTSE)。结果:在40例食管鳞癌中,HPV16的E6、E7基因和CTSE片段的检出率分别是37.5%(15/40)、42.5%(17/40)和40%(16/40),20例正常食管黏膜中E6、E7和CTSE的检出率分别为0%(0/20)、0%(0/20)和5%(1/20),两者均存在显著性差异(P<0.05)。CTSE片段分别与E6和E7基因有明显相关性(P<0.05)。食管鳞癌中E6、E7基因及CTSE的检出率在患者不同性别、年龄、肿瘤浸润程度、淋巴结转移和组织学分级肿瘤中差异均无显著性(P>0.05)。结论:E6和E7基因与CTSE片段共存于HPV16感染的食管鳞癌组织中,三者可能与食管鳞癌的发生和发展有关。
