Polymorphism in the upstream regulatory region of human papilloma virus type 16 from the cervical cancer biopsies in Xinjiang Uygur women

To investigate the mutations in the upstream regulatory region (URR) of human papillomavirus type 16 (HPV-16) from the cervical cancer biopsies in Xinjiang Uygur women and its relationship to the high incidence of cervical cancer in the southern Xinjiang, the tissue DNA was extracted from the cervical cancer biopsies, and the URR segment of HPV-16 DNA was amplified, sequenced and analyzed. Thereafter, the polymorphism of URR in HPV-16 was then analyzed. It was demonstrated that the positive rate detected for the presence of URR in HPV-16 was 89.47% (17/19). Compared with the previously published sequence in URR of prototype HPV-16, some mutations were detected in the sequence of URR. The mutations in 17 URR fragments of HPV-16 could be divided into 11 patterns (XJU-1 to XJU-11) at nucleic acid level, in which each of XJU-1 and XJU-4 accounted for 23.53% (4/17), and other patterns of mutation accounted for 5.88% (1/17) . In comparison with the URR of prototype HPV-16, the DNA identity of these patterns was 98.50%-99.68% . In these 17 URR fragments, two point mutations occurred at position 7192 (G to T) and position 7520 (G to A) and they appeared to be constant in Xinjiang area. These two mutations were ubiquitous in the Asia-American type and conferred strong infection activity and carcinogenicity of this virus. In addition, the mutations at position 7729 (A to C), position 7843 (A to G) and position 7792 (C to T) could enhance its transcription activity considerably. It is concluded that some mutations occur in URR gene of HPV-16 in the cervical cancer biopsies taken from Uygur women in Xinjiang area, suggesting that certain relationship exists among the mutations in URR of HPV-16, the phylogeny of HPV-16 and the high incidence of cervical cancer in southern part of Xinjiang area.

Expression of Bcl-2 and Survivin in uterine cervical carcinogenesis and correlation between the expression and infection of HPV_(16/18)

To investigate the expression of Bcl-2 and Survivin and HPV16/18 infection in uterine cervical carcinogenesis, formalin-fixed, paraffin-embedded specimens from cervical carcinomas, cervical intraepithelial neoplasia （C1N） and normal cervical tissues were studied. Using streptavidin-biotin peroxidase （S- P） immunohistochemical technique, the authors examined the expression of Bcl-2 and Survivin in these specimens. The number of apoptosis cells was assessed in situ by TdT-mediated dUTP-biotin end labeling （TUNEL） method. The infection of HPV type 16, 18 DNA were determined by PCR. It was found that there were significant differences in Bcl-2, Survivin and apoptotic index （AI） between cervical carcinomas, CIN and normal cervical tissues, respectively. Expression of Bcl-2 and AI were correlated with tu- mor grades, clinical stages and lymph node metastasis and expression of Survivin was associated with tu- mor grades and lymph node metastasis. There were different positive rate of HPV^s between cervical car- cinomas, C1N and normal tissues and were not associated with tumor grades, clinical stages and lymph node metastasis. The infection of HPV16/18 was associated with the expression of Bcl-2, Survivin and AI, respectively. It was concluded that the abnormal expression of Bcl-2, Survivin and infection of HPV16/18 were associated with cervical carcinomas. They possibly can be useful indexes for the primary screening and prognosis of cervical carcinomas.

Immune response to plasmid DNA encoding HPV16-L1 protein

Objective To test the immunogenicity of recombinant plasmid DNA containing human papillomavirus type 16 L1 (HPV16 L1) coding sequence of mice Methods The HPV16 L1 encoding sequence was generated by polymerase chain reaction (PCR), and inserted into TA cloning vector PCR Ⅱ, then cloned in the eukaryotic expression vector pcDNA3 1 with CMV promoter The recombinant plasmid DNA pcDNA L1 was transferred into Cos 7 cells and used to immunize BALB/c mice via muscular injection The expression of HPV16 L1 in transferred cells was identified by immunospot and immunocytochemistry, which tested specific anti HPV16 L1 antibody in the serum of immunized mice Results Using the immunospot technique, we found L1 protein expression in pcDNA L1 transferred cells The immunocytochemistry studies demonstrated that the L1 protein was located in nuclei In immunized mice, specific anti HPV16 L1 antibodies could be detected by immunospot and immunocytochemistry 28 days after the first immunization and last at least 41 days Conclusions We constructed HPV16 L1 eukaryotic expressing plasmid whose DNA could induce immuno humoral response in mice This observation will be helpful in designing HPV16 prophylactic vaccine