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1.
Article in Chinese | WPRIM | ID: wpr-826336

ABSTRACT

To explore the molecular mechanism of human papillomavirus subtype 16(HPV-16)E7 oncogene-induced DNA re-replication in response to DNA damage. Flow cytometry was performed to examine the cell cycle changes in RPE1 E7 cells stably expressing HPV-16 E7 and its control cell RPE1 Vector after DNA damage.Immunoblotting assay was used to evaluate the early mitotic inhibitor 1(Emi1)expression in RPE1 E7 and RPE1 Vector cells with or without DNA damage.The changes of the proportion of polyploidy was detected by flow cytometry in DNA-damaged RPE1 E7 cells interfered by Emi1 small interfering RNA. Compared with the control cells,the proportion of polyploids in RPE1 E7 cells was significantly increased in response to DNA damage(=6.397,=0.0031).Emi1 protein expression was significantly increased in DNA damaged RPE1 E7 cells(=8.241,=0.0012).The polyploid ratio of RPE1 E7 cells was significantly reduced after Emi1 was interfered by two independent small interfering RNAs(=2.916,=0.0434;=3.452,=0.0260). In response to DNA damage,Emi1 promoted DNA re-replication caused by HPV-16 E7.


Subject(s)
DNA Damage , DNA Replication , Human papillomavirus 16 , Mitosis , Oncogene Proteins, Viral
2.
Mem. Inst. Oswaldo Cruz ; 115: e190405, 2020. graf
Article in English | LILACS | ID: biblio-1091247

ABSTRACT

BACKGROUND High-risk human papillomaviruses (HR-HPVs) are the etiological agents of cervical cancer. Among them, types 16 and 18 are the most prevalent worldwide. The HPV genome encodes three oncoproteins (E5, E6, and E7) that possess a high transformation potential in culture cells when transduced simultaneously. In the present study, we analysed how these oncoproteins cooperate to boost key cancer cell features such as uncontrolled cell proliferation, invasion potential, and cellular redox state imbalance. Oxidative stress is known to contribute to the carcinogenic process, as reactive oxygen species (ROS) constitute a potentially harmful by-product of many cellular reactions, and an efficient clearance mechanism is therefore required. Cells infected with HR-HPVs can adapt to oxidative stress conditions by upregulating the formation of endogenous antioxidants such as catalase, glutathione (GSH), and peroxiredoxin (PRX). OBJECTIVES The primary aim of this work was to study how these oncoproteins cooperate to promote the development of certain cancer cell features such as uncontrolled cell proliferation, invasion potential, and oxidative stress that are known to aid in the carcinogenic process. METHODS To perform this study, we generated three different HaCaT cell lines using retroviral transduction that stably expressed combinations of HPV-18 oncogenes that included HaCaT E5-18, HaCaT E6/E7-18, and HaCaT E5/E6/E7-18. FINDINGS Our results revealed a statistically significant increment in cell viability as measured by MTT assay, cell proliferation, and invasion assays in the cell line containing the three viral oncogenes. Additionally, we observed that cells expressing HPV-18 E5/E6/E7 exhibited a decrease in catalase activity and a significant augmentation of GSH and PRX1 levels relative to those of E5, E6/E7, and HaCaT cells. MAIN CONCLUSIONS This study demonstrates for the first time that HPV-18 E5, E6, and E7 oncoproteins can cooperate to enhance malignant transformation.


Subject(s)
Humans , Cell Transformation, Viral/genetics , Oncogene Proteins, Viral/metabolism , DNA-Binding Proteins/metabolism , Human papillomavirus 18/metabolism , Oxidation-Reduction , Gene Expression Regulation, Neoplastic , Cell Survival , Cell Line, Tumor/virology , Cell Proliferation
3.
Clinics ; 73(supl.1): e486s, 2018. graf
Article in English | LILACS | ID: biblio-974952

ABSTRACT

Human papillomavirus infection is associated with the development of malignant and benign neoplasms. Approximately 40 viral types can infect the anogenital mucosa and are categorized into high- and low-risk oncogenic human papillomavirus, depending on their association with the development of cervical carcinoma. High-risk human papillomavirus 16 and 18 are detected in 55% and 15% of all invasive cervical squamous cell carcinomas worldwide, respectively. Low-risk human papillomavirus 6 and 11 are responsible for 90% of genital warts and are also associated with the development of recurrent respiratory papillomatosis. Human papillomavirus preferentially infects mitotic active cells of the basal layer from both mucosal and cutaneous epithelium through microabrasions. The viral life cycle synchronizes with the epithelial differentiation program, which may be due, in part, to the binding of differentially expressed cellular transcription factors to the long control region throughout the various epithelial layers. This review aimed to summarize the current knowledge regarding the mechanisms by which viral gene expression is regulated and the influence of human papillomavirus heterogeneity upon this phenomenon. A better understanding of the regulatory mechanisms may elucidate the particularities of human papillomavirus-associated pathogenesis and may provide new tools for antiviral therapy.


Subject(s)
Humans , Papillomaviridae/genetics , Transcription Factors/genetics , Gene Expression Regulation, Viral , Papillomavirus Infections/virology , Papillomaviridae/physiology , Oncogene Proteins, Viral/genetics , Promoter Regions, Genetic/genetics
4.
Braz. j. infect. dis ; 21(3): 248-254, May-June 2017. tab
Article in English | LILACS | ID: biblio-839224

ABSTRACT

ABSTRACT Objective: To evaluate the association between p53 polymorphisms and human papillomavirus (HPV) E6/E7 mRNA expression. Methods: We analyzed 175 cervical samples from women aged 16-69 years old who were tested for HPV E6/E7 mRNA expression (NucliSENS® EasyQ® HPV). The samples were divided into three groups: positive (n = 75) those with positive HPV E6/E7 mRNA expression and positive high-risk HPV Hybrid Capture (HR-HC) test; negative (n = 52) those with negative HPV E6/E7 mRNA expression and positive HR-HC; and control (n = 48) those with negative HPV E6/E7 mRNA expression and negative HR-HC. The p53 polymorphisms at codons 11, 72, and 248 were evaluated through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of the arginine/arginine homozygous genotype at codon 72 was significantly higher in the positive (49.3%) than in the negative (32.7%) and control groups (20.8%, p = 0.002*). The frequency of the arginine allele was also significantly higher in the positive (67.3%) than in the negative (53.8%) and control groups (38.5%, p < 0.001*). The arginine/arginine homozygous genotype was significantly associated with positive HPV E6/E7 mRNA expression (positive group) compared with negative and control groups (odds ratio: 2.633; 95% CI, 1.399-4.954, p = 0.003). The frequency of arginine/arginine homozygous genotype at codon 72 remained significantly more frequent in the positive group of women aged ≥30 years than in the other two groups. Conclusion: The presence of the p53 arginine/arginine homozygous genotype at codon 72 was significantly associated with the positive HPV E6/E7 mRNA expression.


Subject(s)
Humans , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Papillomaviridae/genetics , RNA, Messenger/metabolism , Oncogene Proteins, Viral/genetics , Cervical Intraepithelial Neoplasia/virology , Papillomavirus Infections/virology , Papillomavirus E7 Proteins/genetics , Arginine/genetics , Polymorphism, Restriction Fragment Length , Codon , RNA, Viral , Uterine Cervical Neoplasms/virology , Polymerase Chain Reaction , Tumor Suppressor Protein p53/genetics , Genotype
5.
Rev. bras. ginecol. obstet ; 38(3): 154-159, Mar. 2016. tab
Article in English | LILACS | ID: lil-781452

ABSTRACT

Purpose To correlate the expression of high-risk HPV E6 mRNA with pap smear, colposcopy, and biopsy results in women with high grade squamous intraepithelial lesion (HSIL). Methods A cross-sectional study was performed on women referred for primary care services after cytological diagnosis of HSIL. We evaluated the expression of E6/E7 mRNA of HPV types 16,18,31,33, and 45 and correlated the results with those of Pap smear, colposcopy, and biopsy. For amplification/detection of mRNA E6 / E7 we used NucliSENSEasyQ kit to detect HPV mRNA by polymerase chain reaction with primers/ probes for HPV types 16, 18, 31, 33, and 45. Results Out of 128 valid tests, the results of 30 (23.4%) tests were negative and 98 (70%) tests were positive. Only one type of HPV was detected in 87.7% of the E6/E7 mRNA positive cases. HPV16 was detected in 61.2% of the cases, followed by HPV33 (26.5%), HPV31 (17.3%), HPV18 (10%), and HPV45 (4.08%). Pap smear tests revealed that the E6/E7 test was positive in 107 (83.8%) women with atypical squamous cells - high grade (ASC-H), HSIL, or higher. The E6/E7 test was positive in 69 (57.5%) specimens presenting negative cytology results. When analyzing the association with colposcopy results, the frequency of positive E6/E7 results increased with the severity of the injury, ranging from 57.1% in women without colposcopy-detected injury to 86.5% in those with higher levels of colposcopy findings. Of the 111 women who underwent biopsy and E6/E7 testing, the E6/E7 test was positive in 84.7% of the women who presented with lesions of cervical intraepithelial neoplasia (CIN) grade 2 or higher. Finally, 41.2% of women with a negative biopsy presented a positive E6/E7 test. Conclusions E6/E7mRNA expression was higher in women with HSIL and CIN grade 2 or higher.


Objetivo Correlacionar a expressão mRNAE6/E7 do HPV de alto risco com os exames de Papanicolau, colposcopia e biópsia em mulheres com lesão intraepitelial escamosa de alto grau (HSIL). Métodos Estudo transversal com mulheres encaminhadas aos serviços de atenção primária com diagnóstico citológico de HSIL. Foi avaliada a expressão do mRNAE6/E7 dos tipos de HPV 16,18,31,33 e 45, correlacionando-se a expressão com os exames de Papanicolau, colposcopia e biópsia. Para a amplificação/detecção demRNA de E6/E7 foi usado o kit NucliSENS EasyQ(r) HPV que detecta mRNA do HPV por meio da reação em cadeia da polimerase com primers/probes HPV dos tipos 16, 18, 31, 33 e 45. Resultados Foram obtidos 128 testes válidos. Destes: 30 (23,4%) foram negativos e 98 (70%) dos testes foram positivos. Foi encontrado apenas um tipo de HPV em 87,7% dos positivos. O HPV16 foi omais encontrado em61,2%, seguido pelos HPV33 (26,5%); HPV31 (17,34%); HPV 18 (10,0%) e HPV (45 4,0%). Quanto ao exame de Papanicolau, o teste E6/E7 foi positivo em 107 (83,8%) das mulheres com ASC-H, HSIL ou superior, enquanto em citologia negativa foi encontrado um resultado positivo em 69 (57,5%) colposcopia. A frequência de teste E6/E7 positivo aumentou com a gravidade da lesão, detectada na colposcopia variando de 57,1% em mulheres sem lesão identificada em colposcopia até 86,5% naqueles com achado de colposcopia de grau maior. Das 111 mulheres que se submeteram a biópsia e o teste E6/E7, o teste foi positivo em 84,7% das que apresentaramlesão igual ou superior a NIC 2 (neoplasia intraepitelial cervical) e 41,2% daqueles com biópsia negativa. Conclusões A expressão de E6, E7 RNAm ocorreu commaior frequência emlesões de alto grau citológica e em casos com biópsias de NIC2 ou maior.


Subject(s)
Humans , Female , Pregnancy , Adult , Cervical Intraepithelial Neoplasia/genetics , Oncogene Proteins, Viral/genetics , Squamous Intraepithelial Lesions of the Cervix/genetics , Cross-Sectional Studies , Oncogene Proteins , Papillomaviridae , Papillomavirus Infections/diagnosis , RNA, Messenger/metabolism , Uterine Cervical Neoplasms/diagnosis , Vaginal Smears
6.
Article in English | WPRIM | ID: wpr-258815

ABSTRACT

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>PCR 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.</p><p><b>RESULTS</b>HPV18 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.</p><p><b>CONCLUSION</b>HPV was a contributor to virus caused human breast cancer, suggesting that the oncogenes in HPV were potential targets for gene therapy of breast cancer.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Base Sequence , Breast Neoplasms , Genetics , Therapeutics , Female , Genetic Therapy , Methods , Humans , Middle Aged , Oncogene Proteins, Viral , Genetics , Metabolism , Papillomaviridae , Physiology , Papillomavirus Infections , Genetics , Therapeutics , Sequence Alignment
7.
Protein & Cell ; (12): 575-588, 2015.
Article in English | WPRIM | ID: wpr-757578

ABSTRACT

Human papillomaviruses (HPVs) including high-risk (HR) and low-risk (LR) subtypes have distinguishable variation on both genotypes and phenotypes. The co-infection of multiple HR-HPVs, headed by HPV16, is common in cervical cancer in female. Recently accumulating reports have focused on the interaction between virus and host, particularly the role of human microRNAs (miRNAs) in anti-viral defense by targeting viral genome. Here, we found a well-conserved target site of miRNAs in the genomes of most HR-HPVs, not LR-HPVs, by scanning all potential target sites of human miRNAs on 24 HPVs of unambiguous subtypes of risk. The site is targeted by two less common human miRNAs, miR-875 and miR-3144, and is located in E6 oncogene open reading frame (ORF) and overlap with the first alternative splice exon of viral early transcripts. In validation tests, miR-875 and miR-3144 were identified to suppress the target reporter activity markedly and inhibit the expression of both synthetically exogenous E6 and endogenous E6 oncogene. High level of two miRNAs can inhibit cell growth and promote apoptosis in HPV16-positive cervical cancer cells. This study provides a promising common target of miRNAs for most HR-HPVs and highlights the effects of two low expressed human miRNAs on tumour suppression.


Subject(s)
Apoptosis , Genetics , Base Sequence , Binding Sites , Genetics , Cell Line, Tumor , Cell Proliferation , Genetics , Female , Gene Expression , Host-Pathogen Interactions , Genetics , Human papillomavirus 16 , Genetics , Physiology , Humans , MicroRNAs , Genetics , Microscopy, Fluorescence , Molecular Sequence Data , Oncogene Proteins, Viral , Genetics , Repressor Proteins , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Nucleic Acid , Uterine Cervical Neoplasms , Genetics , Pathology , Virology
8.
Iranian Journal of Cancer Prevention. 2015; 8 (1): 18-23
in English | IMEMR | ID: emr-159983

ABSTRACT

Cervical cancer has been shown to be highly associated with human papillomavirus [HPV] infection. The viral oncogenes E6 and E7 are constantly expressed by the tumor cells and are therefore potent targets for therapeutic genetic vaccination. In the present study, it was investigated the potential effect of HPV-16 E6, E7 and L1 co-administration to activate specific cytotoxic T lymphocytes in tumor mice models. The HPV-16 E6, E7 and L1 genes from Iranian isolate were separately inserted into the mammalian expression vector, pcDNA3, to construct the DNA vaccine candidates. Tumor-bearing Animals [C57BL/6 mice] were immunized with the vaccine candidate; then, Lymphocyte Proliferation Assay [LPA] and relative tumor volume measurements were carried out in order to examine the immunological effects of the vaccine. Obtained results showed that co-administration of the HPV-16 E6, E7 and L1 DNA induced HPV-16 specific cellular immune responses and also protected against TC-1-induced tumor in vivo compared with negative controls. The results showed that mixed delivery systems might be valuable to improve the magnitude of the induced immune responses and confirmed therapeutic effects of HPV-16 E6, E7 through cytotoxic T lymphocyte induction and illustrate the new promising role for HPV-16 L1 CTL epitopes as a suitable CTL inducer


Subject(s)
Animals, Laboratory , Vaccines, DNA , Immunity, Cellular , Oncogene Proteins, Viral , Repressor Proteins , Papillomavirus E7 Proteins , Capsid Proteins , Human papillomavirus 16 , Mice
9.
Iranian Journal of Cancer Prevention. 2015; 8 (6): 35-40
in English | IMEMR | ID: emr-175790

ABSTRACT

Background: The association between human papillomavirus [HPV] infections and cervical cancer has suggested the design of prophylactic and therapeutic vaccines against genital warts. The HPV capsid has made of two L1 and L2 coat proteins that have produced late in viral infections. Regarding to the recent studies, two commercial prophylactic vaccines have based on L1 viral like particles [VLPs] could strongly induce antibody responses, and protect human body from HPV infections. However, the use of these HPV vaccines has hindered due to their high cost and some limitations. Currently, among various vaccination strategies, live vector-based vaccines have attracted a great attention


Objectives: Herein, a non-pathogenic strain of the protozoan organism known as Leishmania tarentolae has utilized to induce potent humoral immunity in mice model


Materials and Methods: At first, cloning of HPV16 L1 gene into Leishmania expression vector has performed and confirmed by PCR and digestion with restriction enzymes. The promastigotes of Leishmania tarentolae [Ltar] have transfected with linearized DNA construct by electroporation. Protein expression has analyzed by SDS-PAGE and western blotting. Then, the immunogenicity of leishmania expressing L1 protein [Ltar-L1] has assessed in mice model


Results: Our data has indicated that subcutaneous immunization of mice with the recombinant L.tar-L1 has led to enhance the levels of IgG1 and lgG2a in comparison with control groups. Furthermore, there was no significant increase in antibody levels between two and three times of immunizations


Conclusions: The recombinant live vector was able to induce humoral immunity in mice without need of any adjuvant. However, further studies have required to increase its efficiency


Subject(s)
Animals, Laboratory , Oncogene Proteins, Viral , Capsid Proteins , Leishmania , Immunity, Humoral , Mice , Antibody Formation
10.
Chinese Journal of Biotechnology ; (12): 566-576, 2015.
Article in Chinese | WPRIM | ID: wpr-240618

ABSTRACT

HPV16 L2E7 is a fusion protein used for therapeutical vaccine targeting HPV virus. To increase its expression in Escherichia coli, we optimized the codon usage of HPV16 l2e7 gene based on its codon usage bias. The optimized gene of HPV16 sl2e7 was cloned into three different vectors: pGEX-5X-1, pQE30, ET41a, and expressed in JM109, JM109 (DE3) and BL21 (DE3) lines separately. A high expression line was selected with pET41a vector in BL21 (DE3) cells. After optimization of the growth condition, including inoculation amount, IPTG concentration, induction time and temperature, the expression level of HPV16 L2E7 was increased from less than 10% to about 28% of total protein. HPV16 L2E7 protein was then purified from 15 L culture by means of SP Sepharose Fast Flow, Q Sepharose Fast Flow and Superdex 200 pg. After renaturing, HPV16 L2E7 protein with ≥ 95% purity was achieved, which was confirmed via SDS-PAGE gel and Western blotting. The combined use of purified HPV16 L2E7 and CpG helper has shown clear inhibition of tumor growth in mice injected with tumor cells, with six out of eight mice shown no sign of tumor. This study lays a solid foundation for a new pipeline of large-scale vaccine production.


Subject(s)
Animals , Capsid Proteins , Codon , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetic Vectors , Human papillomavirus 16 , Mice , Neoplasms, Experimental , Oncogene Proteins, Viral , Papillomavirus E7 Proteins , Papillomavirus Vaccines , Therapeutic Uses , Recombinant Fusion Proteins
11.
Chinese Journal of Virology ; (6): 370-378, 2015.
Article in Chinese | WPRIM | ID: wpr-296274

ABSTRACT

To study the complete genomic sequence, genomic characteristics, and genetic variation of the bovine papillomavirus 2 genotype (BPV-2) Aks-01 strain at the molecular level, genotyping of this strain from the skin samples of cows in southern Xinjiang (China) was first detected by the polymerase chain reaction with FAP59/FAP64 primers. Based on the complete genome of the BPV-2 reference strain, specific primers and sequencing primers were designed, and the complete genome of the Aks-01 strain amplified and sequenced. Sequence analyses showed that genotyping of the Aks-01 strain belonged to BPV-2. The Aks-01 strain had the structural characteristics of BPV-2. The 7944-bp full-length genomic sequence of the Aks-01 strain was compiled using DNAStar™. The sequence of the Aks-01 strain had 98% similarity to the reference strain from GenBank. The Aks-01 strain was most closely related to BPV-1 and BPV-13. BPV-2, BPV-1 and BPV-13 were grouped within the genus Deltapapillomavirus. The Aks-01 strain is the first BPV-2 strain reported in southern Xinjiang.


Subject(s)
Amino Acid Sequence , Animals , Base Sequence , Bovine papillomavirus 1 , Genetics , Cattle , China , Evolution, Molecular , Female , Genome, Viral , Genetics , Genomics , Genotype , Molecular Sequence Data , Oncogene Proteins, Viral , Chemistry , Genetics , Metabolism , Phylogeny , Sequence Analysis, DNA , Skin , Virology
12.
Chinese Journal of Virology ; (6): 530-536, 2015.
Article in Chinese | WPRIM | ID: wpr-296252

ABSTRACT

We wished to screen the cell line that stably expresses the HPV18E5 protein, and to ascertain the influence of HPV18E5 protein on cell proliferation and the cell cycle. The HPV18E5 gene was amplified by the polymerase chain reaction. Then, the His-tag pSecTag-HPV18E5 eukaryotic expression vector was constructed by digestion ligation and connection. The recombinant plasmid was transfected into Balb/c3T3 cells with lipofectamine, and positive cell lines were screened by a culture medium containing bleomycin. HPV18E5 expression in cells was confirmed by western blotting and immuno-enzymatic methods. The influence of HPV18E5 on cell proliferation and the cell cycle were detected by Cell Counting Kit-8 and flow cytometry, respectively. The pSecTag-HPV18E5 eukaryotic expression vector was constructed. After 21-day selection in a culture medium containing 400 μg/mL bleomycin, stably expressing HPV18E5 protein cells were harvested. Compared with control groups, cell proliferation in HPV18E5 stably expressed cells was obviously increased, as was the S phase in the cell cycle. Our results suggested that HPV18E5 influences cell proliferation and the cell cycle. Our study has laid the foundation of the biologic properties of HPV18E5 protein, which will aid further studies on the mechanism of action of carcinogenesis.


Subject(s)
Cell Cycle , Cell Line, Tumor , Cell Proliferation , Human papillomavirus 18 , Genetics , Metabolism , Humans , Oncogene Proteins, Viral , Genetics , Metabolism , Papillomavirus Infections , Virology , Transfection
13.
Rev. latinoam. enferm ; 22(6): 1034-1040, 16/12/2014. tab, graf
Article in English | LILACS, BDENF | ID: lil-732956

ABSTRACT

OBJECTIVES: to identify the number of electro-medical pieces of equipment in a coronary care unit, characterize their types, and analyze implications for the safety of patients from the perspective of alarm fatigue. METHOD: this quantitative, observational, descriptive, non-participatory study was conducted in a coronary care unit of a cardiology hospital with 170 beds. RESULTS: a total of 426 alarms were recorded in 40 hours of observation: 227 were triggered by multi-parametric monitors and 199 were triggered by other equipment (infusion pumps, dialysis pumps, mechanical ventilators, and intra-aortic balloons); that is an average of 10.6 alarms per hour. CONCLUSION: the results reinforce the importance of properly configuring physiological variables, the volume and parameters of alarms of multi-parametric monitors within the routine of intensive care units. The alarms of equipment intended to protect patients have increased noise within the unit, the level of distraction and interruptions in the workflow, leading to a false sense of security. .


OBJETIVOS: identificar o número de alarmes dos equipamentos eletromédicos numa unidade coronariana, caracterizar o tipo e analisar as implicações para a segurança do paciente na perspectiva da fadiga de alarmes. MÉTODO: trata-se de estudo quantitativo observacional descritivo, não participante, desenvolvido numa unidade coronariana de um hospital de cardiologia, com capacidade para 170 leitos. RESULTADOS: registrou-se o total de 426 sinais de alarmes, sendo 227 disparados por monitores multiparamétricos e 199 alarmes disparados por outros equipamentos (bombas infusoras, hemodiálise, ventiladores mecânicos e balão intra-aórtico), nas 40h, numa média total de 10,6 alarmes/hora. CONCLUSÃO: os resultados encontrados reforçam a importância da configuração de variáveis fisiológicas, do volume e dos parâmetros de alarmes dos monitores multiparamétricos à rotina das unidades de terapia intensiva. Os alarmes dos equipamentos destinados a proteger os pacientes têm conduzido ao aumento do ruído na unidade, à fadiga de alarmes, a distrações e interrupções no fluxo de trabalho e à falsa sensação de segurança. .


OBJETIVOS: identificar el número de alarmas de los equipamientos electromédicos en una unidad coronariana, caracterizar el tipo y analizar las implicaciones para la seguridad del paciente en la perspectiva de fatiga de alarmas. MÉTODO: se trata de un estudio cuantitativo, observacional, descriptivo, no participante, desarrollado en una unidad coronariana de un hospital de cardiología, con capacidad de 170 camas. RESULTADOS: se registró un total de 426 señales de alarmas, siendo 227 disparadas por monitores multiparamétricos y 199 disparadas por otros equipamientos (bombas de infusión, hemodiálisis, ventiladores mecánicos y balón intraaórtico), durante 40h, con un promedio total de 10,6 alarmas/hora. CONCLUSIÓN: los resultados encontrados refuerzan la importancia de la configuración de las variables fisiológicas, del volumen y de los parámetros de alarma de los monitores multiparamétricos, a la rutina de las unidades de terapia intensiva. Las alarmas de los equipamientos destinados a proteger a los pacientes, han llevado al aumento del ruido en la unidad, a la fatiga de alarmas, a las distracciones e interrupciones en el flujo de trabajo y a una falsa sensación de seguridad. .


Subject(s)
Humans , DNA-Directed RNA Polymerases/metabolism , Oncogene Proteins, Viral/genetics , RNA Polymerase III/metabolism , Sarcosine/analogs & derivatives , Transcription Factors, TFIII , Transcription, Genetic , Transcription Factors/metabolism , Adenovirus Early Proteins , Detergents , DNA-Binding Proteins/genetics , HeLa Cells , Kinetics , Sarcosine/pharmacology , Transcription Factor TFIIIB , Transcription Factors/genetics , Transcription, Genetic/drug effects
14.
Chinese Journal of Virology ; (6): 201-207, 2014.
Article in Chinese | WPRIM | ID: wpr-356615

ABSTRACT

High-risk human papillomavirus (HPV) is the principal cause of various cancers including cervical cancer, anal cancer, vulvar cancer, and some head and neck cancers. In the viral life cycle, by interacting with both viral and host DNA and proteins, the HPV E2 protein plays a pivotal role in viral transcriptional regulation and DNA replication, and it is also associated with modification of various cellular processes, including host gene transcription, RNA processing, apoptosis, ubiquitination, and intracellular trafficking, to create a convenient environment for a replicative cycle of the virus and contribute to the HPV pathogenesis. Elucidating the roles of E2 protein throughout the viral life cycle will improve our understanding of the viral life cycle and pathogenesis and help us identify novel antiviral agents with therapeutic potential. This article reviews the research progress in the structure, roles, and activity of high-risk HPV E2 protein, particularly that of HPV-16.


Subject(s)
Animals , DNA-Binding Proteins , Genetics , Metabolism , Female , Gene Expression Regulation, Viral , Human papillomavirus 16 , Genetics , Metabolism , Humans , Oncogene Proteins, Viral , Genetics , Metabolism , Papillomavirus Infections , Genetics , Metabolism , Virology , Uterine Cervical Neoplasms , Genetics , Metabolism , Virology
15.
Rev. bras. cancerol ; 59(4): 565-573, out.-dez. 2013. tab
Article in Portuguese | LILACS | ID: lil-724644

ABSTRACT

Introdução: O Papilomavírus Humano (HPV) é um agente epiteliotrópico que apresenta mais de 100 genótipos.Destes, alguns são considerados de alto risco devido ao potencial para induzir o surgimento de lesões malignas, comoo carcinoma cervical, cujo percentual de associação com o referido vírus é de aproximadamente 90%. Nas célulasinfectadas, duas proteínas virais desempenham papel fundamental na tumorigênese. Objetivo: Realizar uma revisão dos trabalhos existentes na literatura científica internacional com enfoque no papel das proteínas virais do HPV na carcinogênese. Método: Os artigos utilizados para a realização da presente revisão foram selecionados e obtidos na íntegra nos portais eletrônicos Pubmed e Periódicos Capes. Os descritores utilizados na busca incluíram: Human Papillomavirus, HPV, viral proteins, E5, E6 e E7. Resultados: As proteínas virais E6 e E7 são amplamente conhecidas por promoverem a degradação das proteínas celulares p53 e pRb, respectivamente, efeito que responde por grande parte do potencial oncogênico dos genótipos de alto risco de HPV, sendo funcionalmente equivalentes a mutações dos referidos genes celulares, que são comumente observadas em diversos tumores. Contudo, novos estudos têm demonstrado que essas proteínas virais também estão envolvidas em diversas outras vias tumorais, denotando novamente a relevância das mesmas nesse processo. Ademais, alguns trabalhos apontam a proteína E5 como coadjuvante na carcinogênese. Conclusão: Os diversos efeitos constatados das proteínas precoces virais culminam no favorecimentoda proliferação celular descontrolada, imortalização, regulação da diferenciação celular, suscetibilidade à metástase e escape da vigilância imunológica


Subject(s)
Humans , Male , Female , Carcinogens , Oncogene Proteins, Viral , Papillomavirus Infections , Papillomaviridae/classification
17.
Article in Chinese | WPRIM | ID: wpr-814893

ABSTRACT

OBJECTIVE@#To investigate the expression of high risk human papilloma virus (HPV) 16-E6 protein in non-neoplastic epithelial disorders of the vulva (NNEDV) and squamous cell carcinoma of the vulva (VSCC), and to explore whether HPV16-E6 protein is the etiological factor in NNEDV and its correlation with squamous cell carcinoma of the vulvae.@*METHODS@#We detected HPV16-E6 protein expression in 15 normal vulvae cases, 40 NNEDV cases and 45 VSCC cases by immunohistochemistry SP method.@*RESULTS@#The positive rate of HPV16-E6 in different vulva tissues: was 0% in the normal vulva, 30% in NNEDV and 66.67% in VSCC, respectively. The overall positive rate and two two comparison had statistical significance. In the NNEDV group, the positive rate of squamous hyperplasia type and lichen sclerosus type was 35% and 25%, respectively, with no statistical significance (P>0.05), but higher than that in the normal vulva skin group (P0.017). The positive rate gradually decreased with the tumor differentiation. The difference in well-differentiated and poorly differentiated, moderately and poorly differentiated had statistical significance (P0.017). The positive rate of lymph node metastasis VSCC was significantly higher than that of non-lymph node metastasis VSCC (P<0.05).@*CONCLUSION@#HPV infection may be an etiological factor for NNEDV. The rise of HPV16-E6 positive rate may be related to the occurrence and development of vulvar squamous cell carcinoma.


Subject(s)
Carcinoma, Squamous Cell , Metabolism , Virology , Female , Humans , Hyperplasia , Oncogene Proteins, Viral , Metabolism , Papillomavirus Infections , Metabolism , Precancerous Conditions , Metabolism , Virology , Repressor Proteins , Metabolism , Vulvar Diseases , Metabolism , Virology , Vulvar Neoplasms , Metabolism , Virology
18.
Article in Chinese | WPRIM | ID: wpr-814813

ABSTRACT

OBJECTIVE@#To explore the relation between human papillomavirus (HPV16) infection and expression of Toll-like receptor 4 (TLR4) in cervical cell lines and cervical lesion tissues and to investigate the effect of TLR4 on cervical cancer progression.@*METHODS@#Expression of HPV16 E6 mRNA was detected by RT-PCR. Western blot and immunohistochemistry were used to detect the expression of TLR4 in H8, SiHa, Caski cell lines and formalin-fixed and paraffin-embedded cervical tissue specimens with cervicitis, cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinama (CSCC). DNA was extracted from paraffin-embedded cervical cancer tissues and HPV16 genes were detected.@*RESULTS@#The differentiation expression of HPV16 E6 mRNA and TLR4 in SiHa and Caski was significantly higher than that of normal cervical cell H8 (P0.05). The expression of TLR4 was significantly correlated with HPV16 infection in CIN and CSCC (r=0.303, P<0.05, r=0.633, P<0.05).@*CONCLUSION@#High expression of TLR4 may play important roles in the development and progression of CIN and CSCC, and the expression of TLR4 can be up-regulated by HPV16 infection.


Subject(s)
Carcinoma, Squamous Cell , Metabolism , Virology , Cell Line, Tumor , Cervical Intraepithelial Neoplasia , Metabolism , Virology , Female , Human papillomavirus 16 , Humans , Immunohistochemistry , Lymphatic Metastasis , Oncogene Proteins, Viral , Metabolism , Papillomavirus Infections , Metabolism , RNA, Messenger , Repressor Proteins , Metabolism , Toll-Like Receptor 4 , Metabolism , Up-Regulation , Uterine Cervical Neoplasms , Metabolism , Virology
19.
Journal of Biomedical Engineering ; (6): 1102-1107, 2013.
Article in Chinese | WPRIM | ID: wpr-259759

ABSTRACT

To construct and express a composite gene vaccine for human papillomavirus 58(HPV58)-associated cervical cancer, we inserted HPV58mE6E7 fusion gene into pCI-Fc-GPI eukaryotic expression vector, constructing a recombinant plasmid named pCI-sig-HPV58mE6E7-Fc-GPI. Then we further inserted fragment of sig-HPV58mE6E7Fc-GPI into the novel vaccine vector PVAX1-IRES-GM/B7, constructing PVAX1-HPV58mE6E7FcGB composite gene vaccine. PVAX1-HPV58mE6E7FcGB vaccine was successfully constructed and identified by restriction endonuclease and sequencing analysis. Eukaryotic expression of fusion antigen sig-HPV58mE6E7-Fc-GPI and molecular ad-juvant GM-CSF and B7. 1 were proved to be realized at the same time by flow cytometry and immunofluorescence. So PVAX1-HPV58mE6E7FcGB can be taken as a candidate of therapeutic vaccine for HPV58-associated tumors and their precancerous transformations.


Subject(s)
Cancer Vaccines , Genetics , Capsid Proteins , Genetics , Female , Humans , Oncogene Proteins, Viral , Genetics , Papillomavirus E7 Proteins , Genetics , Papillomavirus Vaccines , Recombinant Proteins , Genetics , Allergy and Immunology , Uterine Cervical Neoplasms , Vaccines, DNA
20.
Journal of Biomedical Engineering ; (6): 1283-1289, 2013.
Article in Chinese | WPRIM | ID: wpr-259723

ABSTRACT

Hela is the cell line of adenocarcinoma of the uterine cervix, and human papillomavirus (HPV) 18 shows positive. We delivered siRNA with target specifically to HPV18 E7 mRNA into nude mice Hela tumor xenografts by nanopatch to inhibit the HPV gene expression, and further to study the superiority, the best action time and concentration of siRNA of using nanopatch to transfer siRNA in vivo. We designed siRNA that target specifically to HPV18 E7 mRNA (siE7) and checked the effect of siE7 in vitro. Tumor xenografts were transfected with siE7 and GenEscort III by nanopatch. Expression of HPV18 E7 mRNA and protein were detected 0 hours, 24 hours, 48 hours, 72 hours after transfection with PT-PCR and Western blot, and the best action time was analyzed using nanopatch to thansfect siRNA in vivo. We transfected GenEscort III and siE7 of Different concentration into tumor xenografts respectively by nanopatch and intraperitoneal injection. Expression of HPV18 E7 mRNA and protein was detected 72 hours after transfection by PT-PCR and Western blot, to analyze the best action concentration of siRNA and the superiority of using nanopatch to thansfect siRNA in vivo. The results proved that SiE7was efficient to inhibit expression of HPV18 E7 mRNA and to advance Hela apoptosis in vitro. SiE7 transfected by nanopatch into xenografts could inhibit effectively expression of HPV18 E7 mRNA and protein. The best action time and concentration of siRNA of using nanopatch to thansfect siRNA in vivo are 72 hour post-transfection and 2 micromol/L siE7. To compare intraperitoneal injection in delivering siRNA in vivo, the effect of nanopatch is very predominant. It can be well concluded that Nanopatch can effectively transfer siRNA in vivo, which can effectively inhibit the HPV gene expression.


Subject(s)
Animals , Apoptosis , DNA-Binding Proteins , Genetics , Female , Gene Expression Regulation, Viral , HeLa Cells , Humans , Mice , Mice, Nude , Nanostructures , Oncogene Proteins, Viral , Genetics , Papillomaviridae , Genetics , RNA, Small Interfering , Transfection , Uterine Cervical Neoplasms , Therapeutics , Xenograft Model Antitumor Assays
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