With the development of biomolecular technology, more human papillomavirus (HPV) types have been found to be related with various diseases. High-risk HPV 16, 18, 31, 33, 45, 52, and 61 have shown causal relationship with cancers of the cervix, vulva, vagina, penis, anus, oral cavity, and oropharynx. Low-risk HPV 6 and 11 have shown causal relationship with genital warts. HPV may also cause cancer of the larynx. However, the relationships between HPV and esophagus and lung cancers are still controversial.
Humans ; Neoplasms ; virology ; Papillomaviridae ; physiology

AIMTo pretreat sperm at various temperatures before exposure to human papillomavirus (HPV) 16 DNA fragments and to assess the efficiency of HPV carrier sperm to transfect cumulus cells.

METHODSCumulus cells from follicular aspirates were obtained, pooled and divided into culture dishes containing Sybr Gold-stained HPV DNA carrying sperm that were either pretreated at 4 degree C, 37 degree C or 40 degree C (n = 5). The cells were incubated in 5% CO(2) in air mixture at 37 degree C for 24 hours. The efficiency of sperm to take up fluorescent HPV DNA was determined at hour 0. After incubation, cumulus cell viability was assessed using the eosin method and the percentages of fluorescent cumulus cells determined.

RESULTSOver half of all the cumulus cells became fluorescent with the highest percentage in the 37 degree C group. Sperm pretreated at 4 degree C had the greatest amount of HPV DNA fragments. Total sperm motility was similar for the 3 pretreatment groups. There were no differences in cumulus viability among the groups.

CONCLUSIONSperm pretreated at 37 degree C transferred the greatest amount of fluorescent HPV DNA fragments to the cumulus cells. The HPV DNA was observed in the nuclear and cytoplasmic compartments. The data suggested the possibility of sperm as a vector for the transmission of HPV DNA to the cumulus cells surrounding ovulated oocytes, which might lead to early implantation failures.

DNA, Viral ; pharmacokinetics ; Humans ; Male ; Papillomaviridae ; genetics ; Spermatozoa ; physiology ; Temperature ; Transfection ; methods

Common warts are close associated with HPVs infection. In this study, we amplified and sequenced the LCR fragment and E2 gene of HPV-2 that infected the patient of extensive common wart with cutaneous horns, and we constructed the recombinant CAT-reporter plasmids pBLCAT-LCR containing HPV-2 prototype or variant LCR and mammalian expression plasmids pcDNA3. 1-E2 containing prototype or variant E2 ORF individually. The promoter activities of HPV-2 variant and the transcriptional repression activities of the mutated E2 protein were evaluated by transient transfection into HeLa cells. The results showed that there were several mutations in LCR and E2 gene of HPV-2 variant. Compared with the prototype, the viral early promoter activity of variant was significantly increased uder the control of LCR. Compared with the wild type E2 protein, the transcriptional repression activities of the mutated E2 protein was abolished partially. We speculate herein that increased promoter activities and decreased repression effect of the mutated E2 protein are linked, at least partially, with the clinical phenotypes of the uncommon huge common wart.
DNA-Binding Proteins ; genetics ; physiology ; Humans ; Mutation ; Oncogene Proteins, Viral ; genetics ; physiology ; Papillomaviridae ; genetics ; Promoter Regions, Genetic ; Repressor Proteins ; physiology ; Warts ; virology

Active Transport, Cell Nucleus ; Capsid Proteins ; physiology ; Heparan Sulfate Proteoglycans ; physiology ; Humans ; Oncogene Proteins, Viral ; physiology ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; etiology ; Viral Structural Proteins ; physiology ; Virion ; physiology ; Virus Assembly

PURPOSE: Human papillomavirus (HPV) infection is a well-known cause of cervical cancer, which, along with its precursors, can be diagnosed and treated with cervical conization (CC). This study aimed to assess HPV- and procedure-related knowledge among women who had undergone CC. MATERIALS AND METHODS: Between February and May 2014, consecutive women who had undergone CC at five different educational hospitals were recruited. All patients had undergone a loop electrosurgical excision procedure as the method of CC. A survey was conducted with a self-developed, 29-item questionnaire, measuring knowledge related to HPV and CC. We analyzed the responses of 160 patients who completed the questionnaire. RESULTS: Mean total knowledge scores (±standard deviation) for HPV and CC were 5.2±3.0 of a possible 13.0 and 8.3±4.2 of a possible 16.0, respectively. While 73% of the patients knew that HPV is the main cause of cervical cancer, only 44% knew that HPV is sexually transmitted. The purpose of CC was correctly identified by 71% of the patients. However, 35% failed to indicate the anatomical area resected at the time of CC in the schematic diagram. Women who were younger (p<0.001), had higher education level (p<0.001), and higher family income (p=0.008) had higher knowledge scores. In contrast, neither interval from CC to survey nor disease severity were associated with total knowledge score. CONCLUSION: The level of knowledge related to HPV and CC was unexpectedly low in women who had undergone CC. Intuitive educational resources may improve this knowledge, and further cohort studies are warranted.
Adult ; Cervix Uteri/*pathology/*virology ; *Conization ; Female ; *Health Knowledge, Attitudes, Practice ; Humans ; Middle Aged ; Papillomaviridae/*physiology ; Republic of Korea ; Sexual Behavior ; *Surveys and Questionnaires ; Uterine Cervical Neoplasms/virology ; Young Adult

Human papillomavirus (HPV) is a common small DNA tumor virus that specifically infects squamous epithelial cells and causes benign or malignant epithelial lesions such as genital warts and cervical cancer. High-risk HPV is detected in specimens of more than 90% of cervical cancer. In the 7. 9 kb genome of HPV, E6 and E7 are the crucial viral oncoproteins that consistently maintained after viral integration into host cell genome. These two proteins interfere with cell proliferation and differentiation through interacting with important tumor suppressors including p53 and pRb. High-risk HPV E6/E7 also induces genomic instability, facilitating cell transformation.
Cell Differentiation ; Cell Proliferation ; Cell Transformation, Neoplastic ; metabolism ; pathology ; Female ; Genomic Instability ; Host-Pathogen Interactions ; Humans ; Oncogene Proteins, Viral ; genetics ; physiology ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; metabolism ; pathology ; Uterine Cervical Neoplasms ; metabolism ; pathology ; virology

To prepare carboxyl terminus truncated human papillomavirus type 58 L1 (HPV58L1) protein and evaluate its ability to form virus-like particles, the baculovirus and Sf-9 insect cells was used to express HPV58L1 protein, and pFastBac-Htb containing HPV58L1 gene sequence of carboxyl terminus truncation was generated. Then Sf-9 cells were infected with recombinant baculovirus. After being cultured, the post-infected cells expressing--HPV58L1 protein were harvested and analyzed by SDS-PAGE and Western blot. The ProBond purification system was used for protein purification. The bio-activity of purified protein was identified by mouse erythrocyte hemagglutination assay, and the VLP formation was examined with transmission electron microscope. Our results showed that the recombinant baculovirus was generated and the Sf-9 cells was infected with the recombinant baculovirus, and after collecting, total cellular proteins were extracted. Truncated HPV58L1 protein with MW 58KD was revealed by SDS-PAGE and confirmed by Western blot. The purified L1 proteins under native condition could cause mouse erythrocytes to agglutinate and form VLP. It is concluded that HPV58L1 protein with carboxyl terminus truncation could be efficiently expressed. In baculovirus Sf-9 cells expression system, the purified protein could self-assemble into virions in vitro, and induce agglutination of mouse erythrocytes, indicating that carboxyl terminus truncation does not interfere with the bioactivity of HPV58L1 protein.
Baculoviridae ; genetics ; metabolism ; Capsid Proteins ; Carbon Dioxide ; Humans ; Inclusion Bodies, Viral ; ultrastructure ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; immunology ; Papillomaviridae ; physiology ; Recombinant Proteins ; biosynthesis ; Virion ; growth & development ; physiology ; Virus Assembly

OBJECTIVETo investigate the correlation between the presence of human papillomavirus (HPV) DNA in the lymph nodes and histopathologically confirmed metastasis of early-stage cervical carcinoma.

METHODSHPV L1 gene fragment in paraffin-embedded tissue specimens of the primary tumor and pelvic lymph nodes from 31 patients with cervical cancer was amplified using HPV-specific PCR with general consensus primers GP5+/GP6+. The type of HPV was identified by sequence analysis of the PCR products, and the correlation between the presence of HPV DNA in the lymph node and the clinicopathological indices of cervical carcinoma was analyzed.

RESULTSThe positivity rate of HPV DNA in the pelvic lymph nodes was 58.1% in the 31 patients, and in 13 of the patients with confirmed metastasis, the detection rate was 84.6% as compared with the rate of 27.8% in the other 18 patients without metastases. The presence of HPV DNA in the lymph node was associated with histologically confirmed metastases. The results of both HPV DNA detection and pathological examination indicated that the obturator, internal iliac and external iliac lymph nodes were more liable to be positive for HPV DNA, accounting for over 90% of the positivity.

CONCLUSIONHPV DNA detection in the pelvic lymph nodes is a helpful predictive factor of metastases, and the obturator, internal iliac and external iliac lymph nodes are the among the most vulnerable lymph nodes of metastatic involvement by early-stage cervical carcinoma.

Adult ; DNA, Viral ; analysis ; genetics ; Female ; Host-Pathogen Interactions ; Humans ; Lymph Nodes ; pathology ; virology ; Lymphatic Metastasis ; Middle Aged ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; pathology ; virology ; Polymerase Chain Reaction ; Tumor Virus Infections ; pathology ; virology ; Uterine Cervical Neoplasms ; pathology ; virology

OBJECTIVETo explore the viral etiology of human breast cancer to determine whether there are novel molecular targets for gene therapy of breast cancer and provide evidence for the research of gene therapy and vaccine development for breast cancer.

METHODSPCR was used to screen HPV16 and HPV18 oncogenes E6 and E7 in the SKBR3 cell line and in 76 paraffin embedded breast cancer tissue samples. RNA interference was used to knock down the expression of HPV18 E6 and E7 in SKBR3 cells, then the changes in the expression of cell-cycle related proteins, cell viability, colony formation, metastasis, and cell cycle progression were determined.

RESULTSHPV18 oncogenes E6 and E7 were amplified and sequenced from the SKBR3 cells. Of the patient samples, 6.58% and 23.68% were tested to be positive for HPV18 E6 and HPV18 E7. In the cell culture models, the knockdown of HPV18 E6 and E7 inhibited the proliferation, metastasis, and cell cycle progression of SKBR3 cell. The knockdown also clearly affected the expression levels of cell cycle related proteins.

CONCLUSIONHPV was a contributor to virus caused human breast cancer, suggesting that the oncogenes in HPV were potential targets for gene therapy of breast cancer.

Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Breast Neoplasms ; genetics ; therapy ; Female ; Genetic Therapy ; methods ; Humans ; Middle Aged ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomaviridae ; physiology ; Papillomavirus Infections ; genetics ; therapy ; Sequence Alignment

OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication