Most oral squamous cell carcinoma (OSCC) is believed to develop via a multistep process of cumulative gene damage in epithelial cells. Increasing incidence of OSCC and evidence that traditional risk factors may not be responsible directed us to investigate the prevalence of virus in pre- and malignant samples.The integration of the DNA from human papillomavirus (HPV), Epstein-Barr virus (EBV), and Herpes simplex (HSV) into the human genome is associated with the expression of oncogenes and the down-regulation of tumor-suppressor genes in OSCC carcinogenesis. This thesis compared samples from India and Sudan, two countries on two continents having a documented high incidence of oral cancer, with specimens from Sweden, with its known low incidence of oral cancer. Each region has, in addition to smoking, a unique non-smoked tobacco habits with documented carcinogenic effects. These countries also typify areas of low and high socioeconomic living conditions with their expected impact on disease development. The study populations were selected from tobacco users and nonusers with OSCC, oral sub-mucous fibrosis (India), oral lichen planus (Sweden), oral leukoplakia with and without dysplasia and snuff-induced lesions (Sweden and Sudan). An expedient method was developed for extracting DNA from old formalin-fixed and paraffin-embedded biopsies. The prevalence of HPV, EBV, and HSV was investigated using PCR/DNA sequencing and southern blot hybridization analysis. We found HPV and EBV to be most prevalent in samples of tissue characterized as normal, with decreasing prevalence in dysplastic and malignant lesions. This intriguing finding that prevalence decreases as neoplastic development proceeds warrants further investigation. Our data do not at first sight support the conclusion that viruses and tobacco use jointly interact with cell mechanisms in the development of oral cancer.

L'infection au papillomavirus humain (HPV) est connue pour être le principal facteur causal d'une série de maladies aussi bien bénignes (condylomatose ano-génitale, papillomatose lyringée, et autres) que malignes (cancer du col de l'utérus, certains cancers ORL, et autres). Deux vaccins prophylactiques (Gardasil® et Cervarix®) sont sur la marché depuis à peu près quatre ans pour prévenir cette infection. Le présent travail de thèse comportait trois objectifs principaux : i) étudier in-silico l'immunogénicité du vaccin Gardasil® ; ii) étudier in-silico l'histoire naturelle d'une infection à HPV et iii) évaluer in-silico le potentiel de l'hypothèse thérapeutique suivante : l'administration intramusculaire du vaccin Gardasil® chez des patients atteints d'une papillomatose laryngée induirait un effet bénéfique car l'arrivée des immunoglobulines au tissu affecté empêcherait l'HPV de compléter son cycle de vie et, par conséquent, la maladie de se propager. Les principales conclusions sont : i) pour qu'une papillomatose laryngée ne s'étende pas il faudrait, d'après nos simulations, que le taux d'IgGs sériques soit maintenu au-dessus de 200 mMU/mL ; ii) pour rester, sur une période de 10 ans, le plus longtemps possible au-dessus de ce seuil (d'effet thérapeutique), en administrant la quantité minimale de vaccin, il faudrait, d'après nos simulations, suivre le protocole suivant : l'immunisation de base (à 0, 2 et 6 mois), suivie de trois rappels successifs tous les six mois jusqu'au 24ème mois, suivis d'un rappel 18 mois plus tard ; iii) par ailleurs, il semble inutile (voire contreproductif), d'après nos simulations, de modifier le schéma traditionnel de base (0-2-6 mois)

<p>In this thesis techniques are presented that aim to determine individual DNA signatures by controlled synthesis of nucleic acid multimers. Allele-specific extension reactions with an improved specificity were applied for several genomic purposes. Since DNA polymerases extend some mismatched 3’-end primers, an improved specificity is a concern. This has been possible by exploiting the faster extension of matched primers and applying the enzymes apyrase or Proteinase K. The findings were applied to methods for resequencing and viral and single nucleotide polymorphism (SNP) genotyping.</p><p>P53 mutation is the most frequent event in human cancers. Here, a model system for resequencing of 15 bps in p53 based on apyrase-mediated allele-specific extension (AMASE) is described, investigated and evaluated (Paper I). A microarray format with fluorescence detection was used. On each array, four oligonucleotides were printed for each base to resequence. Target PCR products were hybridized and an AMASE-reaction performed in situ to distinguish which of the printed oligonucleotides matched the target. The results showed that without the inclusion of apyrase, the resulting sequence was unreadable. The results open the possibilities for developing large-scale resequencing tools.</p><p>The presence of certain types of human papillomaviruses (HPV) transforms normal cells into cervical cancer cells. Thus, HPV type determination is clinically important. Also, multiple HPV infections are common but difficult to distinguish. Therefore, a genotyping platform based on competitive hybridization and AMASE is described, used on clinical sample material and evaluated by comparison to Sanger DNA sequencing (Papers II and III). A flexible tag-microarray was used for detection and the two levels of discrimination gave a high level of specificity. Easy identification of multiple infections was possible which provides new opportunities to investigate the importance of multiply infected samples.</p><p>To achieve highly multiplexed allele-specific extension reactions, large numbers of primers will be employed and lead to spurious hybridizations. Papers IV to VI focus on an alternative approach to control oligomerization by using protease mediated allele-specific extension (PrASE). In order to maintain stringency at higher temperatures, Proteinase K, was used instead of apyrase, leading to DNA polymerase degradation and preventing unspecific extensions. An automated assay with tag-array detection for SNP genotyping was established. First PrASE was introduced and characterized (Paper IV), then used for genotyping of 10 SNPs in 442 samples (Paper V). A 99.8 % concordance to pyrosequencing was found. PrASE is a flexible tool for association studies and the results indicate an improved assay conversion rate as compared to plain allele-specific extension.</p><p>The highly polymorphic melanocortin-1 receptor gene (MC1R) is involved in melanogenesis. Twenty-one MC1R variants were genotyped with PrASE since variants in the gene have been associated to an increased risk of developing melanoma. A pilot study was performed to establish the assay (Paper VI) and subsequently a larger study was executed to investigate allele frequencies in the Swedish population (Paper VII). The case and control groups consisted of 1001 and 721 samples respectively. A two to sevenfold increased risk of developing melanoma was observed for carriers of variants.</p>

Les HPV sont les agents d'infections latentes, d'hyperplasies bénignes ou encore de cancer. Afin de mieux comprendre leur pathogenèse, nous avons cartographié les réseaux d'interactions de protéines de régulation virale E2 pour 12 génotypes HPV. Par double hybride, nous avons procédé à l'identification des interacteurs des protéines E2 suivi par une validation en cellule de mammifère par une méthode basée sur la reconstitution d'une luciferase. Le regroupement des profiles d'interaction montre une corrélation avec la phylogénie, établissant ainsi la contribution de E2 dans la pathogénie associée aux HPV. L'étude des réseaux d'interaction a révélé le ciblage préférentiel de protéines cellulaires hautement connectées, impliquées dans 5 catégories fonctionnelles récapitulant les principales fonctions de E2 mais aussi ouvrant de nouvelles perspectives quant au rôle de cette protéine virale dans les mécanismes d'infection. Dans un deuxième temps, ce travail s'est focalisé sur l'étude d'une interaction impliquant spécifiquement la protéine E2 d'HPV16, le virus le plus représenté dans les cancers cervicaux, et une protéine cellulaire, CCHCR1. En identifiant la surface d'interaction de CCHCR1 sur E2, il s'est avéré qu'elle induisait une compétition avec BRD4, un interacteur majeur de E2, se traduisant par une diminution des capacités transcriptionelle de E2. De même, nous avons montré que CCHCR1 induisait la délocalisation de E2 du noyau vers le cytoplasme. Enfin, nos résultats indiquent qu'en présence de CCHCR1, HPV16 E2 est moins apte à induire la différentiation précoce des kératinocytes, ce qui pourrait potentiellement avoir un effet important sur le cycle viral.

This dissertation observes the association between human papillomavirus (HPV) integration into the host cell genome and cervical intraepithelial lesions. The aim of this study is to determine how the grade of HPV16 DNA integration into the host cell genome (HPV E2 gene deletion) is related to cervical intraepithelial lesions. 253 women were screened for HPV infection and the majority was diagnosed with HPV type 16. The most frequently determined was HPV grade II integration. No statistically significant difference was defined while analyzing the relations between the status of HPV E2 gene integration and the grade of cervical intraepithelial lesions. The fact that integration of HPV type 16 was determined in low grade cervical squamous intraepithelial lesions or in cases with no intraepithelial lesions shows that virus integration occurs at the early stage of carcinogenesis. It is noteworthy that during the follow-up (secondary visit after 6 month) 50% of HPV positive women were identified with mRNR expression which probably establishes the presence of persistent active HPV infection. The research provided is highly significant for improvement of cervical cancer screening programms. Adjusting Pap smear, HPV DNA detection and determination of other HPV biomarkers (such as E2 gene deletion mRNR), it becomes possible to separate out HPV positive women with no clinical signs, although, belonging to the of high risk group for cervical carcinoma developing. / Disertacijoje nagrinėjama sąsaja tarp žmogaus papilomos viruso (ŽPV) integracijos į gimdos kaklelio epitelio ląstelių genomą ir intraepitelinių gimdos kaklelio pokyčių. Pagrindinis darbo tikslas – nustatyti, kaip 16 tipo ŽPV DNR integracijos laipsnis (ŽPV E2 geno iškrita) susijęs su intraepiteliniais gimdos kaklelio pokyčiais. Ištyrus 253 moteris nustatyta, kad dauguma moterų buvo infekuotos 16 tipo ŽPV. Dažniausiai nustatyta II laipsnio šio tipo ŽPV integracija. Statistiškai reikšmingo skirtumo analizuojant sąsajas tarp ŽPV E2 geno integracijos pobūdžio ir intraepitelinių gimdos kaklelio pokyčių laipsnio nenustatyta. Tai, kad integruotų 16 tipo ŽPV formų nustatyta esant nežymių arba net nesant intraepitelinių gimdos kaklelio pokyčių, rodo, jog viruso integracija yra ankstyvasis kancerogenezės įvykis. Pažymėtina, kad pakartotinio patikrinimo metu 50 proc. ŽPV infekuotų moterų konstatuota mRNR raiška, kas rodo besitęsiančią aktyvią ŽPV infekciją. Atliktas tyrimas yra svarbus gerinant patikros dėl gimdos kaklelio patologijos programas: derinant Pap testą ir ŽPV DNR bei kitų ŽPV žymenų nustatymą (mRNR, E2 geno iškrita) galima atrinkti infekuotas ŽPV moteris, kurioms dar nėra klinikinių požymių, tačiau jos priklauso didelės rizikos susirgti gimdos kaklelio vėžiu grupei.

Disertacijoje nagrinėjama sąsaja tarp žmogaus papilomos viruso (ŽPV) integracijos į gimdos kaklelio epitelio ląstelių genomą ir intraepitelinių gimdos kaklelio pokyčių. Pagrindinis darbo tikslas – nustatyti, kaip 16 tipo ŽPV DNR integracijos laipsnis (ŽPV E2 geno iškrita) susijęs su intraepiteliniais gimdos kaklelio pokyčiais. Ištyrus 253 moteris nustatyta, kad dauguma moterų buvo infekuotos 16 tipo ŽPV. Dažniausiai nustatyta II laipsnio šio tipo ŽPV integracija. Statistiškai reikšmingo skirtumo analizuojant sąsajas tarp ŽPV E2 geno integracijos pobūdžio ir intraepitelinių gimdos kaklelio pokyčių laipsnio nenustatyta. Tai, kad integruotų 16 tipo ŽPV formų nustatyta esant nežymių arba net nesant intraepitelinių gimdos kaklelio pokyčių, rodo, jog viruso integracija yra ankstyvasis kancerogenezės įvykis. Pažymėtina, kad pakartotinio patikrinimo metu 50 proc. ŽPV infekuotų moterų konstatuota mRNR raiška, kas rodo besitęsiančią aktyvią ŽPV infekciją. Atliktas tyrimas yra svarbus gerinant patikros dėl gimdos kaklelio patologijos programas: derinant Pap testą ir ŽPV DNR bei kitų ŽPV žymenų nustatymą (mRNR, E2 geno iškrita) galima atrinkti infekuotas ŽPV moteris, kurioms dar nėra klinikinių požymių, tačiau jos priklauso didelės rizikos susirgti gimdos kaklelio vėžiu grupei. / This dissertation observes the association between human papillomavirus (HPV) integration into the host cell genome and cervical intraepithelial lesions. The aim of this study is to determine how the grade of HPV16 DNA integration into the host cell genome (HPV E2 gene deletion) is related to cervical intraepithelial lesions. 253 women were screened for HPV infection and the majority was diagnosed with HPV type 16. The most frequently determined was HPV grade II integration. No statistically significant difference was defined while analyzing the relations between the status of HPV E2 gene integration and the grade of cervical intraepithelial lesions. The fact that integration of HPV type 16 was determined in low grade cervical squamous intraepithelial lesions or in cases with no intraepithelial lesions shows that virus integration occurs at the early stage of carcinogenesis. It is noteworthy that during the follow-up (secondary visit after 6 month) 50% of HPV positive women were identified with mRNR expression which probably establishes the presence of persistent active HPV infection. The research provided is highly significant for improvement of cervical cancer screening programms. Adjusting Pap smear, HPV DNA detection and determination of other HPV biomarkers (such as E2 gene deletion mRNR), it becomes possible to separate out HPV positive women with no clinical signs, although, belonging to the of high risk group for cervical carcinoma developing.

Most epidemiologic studies of sexually transmitted diseases (STD) are based on patients seeking help or advice at various health care settings. Because many STD:s are subclinical, epidemiologic surveys can be strengthened by a population-based approach. The aims of the present study were to investigate the prevalence and incidence of STDs in a population of young women, and to assess associations between STDs and social background, education, previous genital infections, sexual behaviour, contraceptive use and reproductive experience. All women belonging to the 19-, 21-, 23- and 25-year age cohorts and living in the catchment area of a community health center, were invited by mail to participate in the study. In the presence of the investigator, participants answered a structured questionaire regarding their social background, education, previous genital infections, sexual behaviour, contraceptive use and reproductive experience. A gynecologic examination was performed. Cervical scrapes for human papillomavirus (HPV) DNA, as well as samples for wet smear, cervical pap smear, and Chlamydia trachomatis (CT) culture were taken. The presence of genital warts was noted, and a colposcopy was performed 2-5 minutes after application of 5% acetic acid on the cervix and vulva. Acetowhite changes were then assessed. A serologic test for CT and herpes simplex virus type 2 (HSV-2) antibodies were performed. Of the 816 women available, 611 (75%) participated in the study. One out of four women reported symptoms from the lower genital tract. The most common were itching, followed by discharge and soreness. There was a significant correlation between the womens" complaint of vaginal discharge, and previous CT infection, lack of lactobacilli and the presence of leucocytosis in wet smear. Twenty-two percent of the women were HPV DNA positive and acetowhitening at the cervix was observed in 16% of the women. The sensitivity of detection of HPV infection by acetowhitening of the cervix was 22% (95%CI 18%, 26%), and the specificity was 90% (95% Cl 87%, 93%). C.trachomatis culture positivity was found in 2.7% of the women and the seroprevalence of CT was 24.7 %. Atypical cytology was found in 3.4% of the women and 6.6% was HSV-2 seropositiv. Of the women studied 23.6% reported having had at least one STD previously and the laboratory analysis showed 45.4% to have had at least one STD. Multivariate logistic regression analysis showed that the number of sexual partners, age at first coitus, history of therapeutic abortion, and previous pelvic inflammatory disease (PID) was independently correlated with CT seropositivity. Lifetime number of sexual partners was the only independent risk factor for HPV. Multivariate analysis showed that increasing age, early sexual debut, and a history of spontaneous abortion were independently related to the presence of HSV-2 antibodies. The lifetime number of sexual partners and coitus on first date were independently associated with a previous STD. Conclusion, We found that one out of four women had some kind of lower genital tract complaint, almost every other women had at sometime in their life an STD, and STDs were often asymptomatic. Acetowhitening of the cervix and vulva has low sensitivity, to low to warrant its use as a predictor of subclinical HPV infection. The pattern of risk factors differed between STDs. / <p>Härtill 6 uppsatser</p> / digitalisering@umu

HPV疫苗一方面是新興醫療科技產物，另一方面牽涉科學風險與社會影響力未知的議題，是當前醫療科技風險即具代表性的例子。在各利害關係人的論述競逐下，媒體如何建構此類新聞鮮少有研究關注。因此，本研究的主要目的即在探討台灣與美國媒體如何建構人類乳突病毒疫苗（簡稱HPV疫苗）爭議新聞內涵，而在地社會文化特色是否影響傳播文本的疫苗風險知識建構與定義。此外，閱聽眾對新藥的認知受媒體報導內容影響，本研究同時將疫苗風險、利益與侷限性等資訊納入分析標的，探討台灣與美國媒體HPV疫苗新聞品質。 本研究以2003年5月1日《蘋果日報》發刊為起點，至2009年6月30日為止，《聯合報》、《蘋果日報》、《紐約時報》與《今日美國》共245則HPV疫苗新聞為樣本，採用內容分析法檢視四報的HPV疫苗新聞有何特性，如何呈現HPV疫苗風險與利益資訊。 研究結果發現，HPV疫苗新聞的版面會因報紙類別、媒體屬性呈現差異，《聯合報》、《蘋果日報》、《今日美國》的HPV疫苗新聞較常出現在「生活版」或「健康」版；《紐約時報》則是以「焦點／綜合」版和「評論」版為主。且菁英報較常以硬性新聞取徑報導HPV疫苗新聞，通俗報則多以生活、消費情報處理之。在新聞形式方面，《紐約時報》與《今日美國》的「評論性文章」較其他兩報多，而通俗報出現以「專題／特寫／專訪」提供HPV疫苗爭議深度報導的趨勢。在新聞篇幅方面，《紐約時報》「1001字以上」的報導最多，《蘋果日報》以「601-1001字」為主，其於兩報多為「301-600字」的報導，但新聞篇幅不因媒體屬性而有異。 各報對HPV疫苗新聞的重要新聞主題選擇小有差異，但皆以「科學與醫學資訊」類和「政策法規」類主題的報導最多。此外，菁英報較重視政策爭議，但通俗報較聚焦科學研究成果。而台灣地區報導關注「喚醒疾病防治意識」類主題，但美國地區則企圖以「喚醒用藥安全意識」類報導提醒民眾HPV疫苗存在的未知風險。但HPV疫苗主要新聞主題呈現不受報導時間影響。 HPV疫苗新聞消息來源分布會因報導地區而出現差異。雖然「科學界人士」為各時期、各報引用傾向最高的消息來源，但台灣媒體提供「科學界人士」與「政府官員」較多的HPV疫苗議題發言權；美國媒體則是讓「使用者／民眾」有較多的發言空間，並較台灣媒體更常刊出HPV疫苗相關社論。此外，美國媒體較常註明消息來源與利害關係人的互動狀況。 至於新聞框架方面，整體而言，「科學創新框架」與「醫學藥品框架」為報導比例最高的二個新聞框架，但重要性隨時間變化遞減，然「希望框架」和「質疑科學框架」則是出現相反趨勢。此外，菁英報刊出「質疑科學框架」的比例亦高於通俗報。再者，美國媒體較常以「質疑科學框架」、「性／文化框架」建構HPV疫苗新聞；台灣媒體則偏好「希望框架」。 疫苗風險與利益資訊呈現各項目受報紙類別、媒體屬性與報導地區的影響不一。首先，《蘋果日報》提供較多關於「僅預防特定型別」、「抹片檢查」與「安全性行為」的資訊，而通俗報出現「僅預防特定型別」與「安全性行為」兩項目的比例較菁英報高。此外，美國媒體報導「疫苗利益」、「疫苗風險」、「僅預防特定型別」、「施打疫苗後仍須進行抹片檢查」的比例較台灣媒體高。

In this thesis techniques are presented that aim to determine individual DNA signatures by controlled synthesis of nucleic acid multimers. Allele-specific extension reactions with an improved specificity were applied for several genomic purposes. Since DNA polymerases extend some mismatched 3’-end primers, an improved specificity is a concern. This has been possible by exploiting the faster extension of matched primers and applying the enzymes apyrase or Proteinase K. The findings were applied to methods for resequencing and viral and single nucleotide polymorphism (SNP) genotyping. P53 mutation is the most frequent event in human cancers. Here, a model system for resequencing of 15 bps in p53 based on apyrase-mediated allele-specific extension (AMASE) is described, investigated and evaluated (Paper I). A microarray format with fluorescence detection was used. On each array, four oligonucleotides were printed for each base to resequence. Target PCR products were hybridized and an AMASE-reaction performed in situ to distinguish which of the printed oligonucleotides matched the target. The results showed that without the inclusion of apyrase, the resulting sequence was unreadable. The results open the possibilities for developing large-scale resequencing tools. The presence of certain types of human papillomaviruses (HPV) transforms normal cells into cervical cancer cells. Thus, HPV type determination is clinically important. Also, multiple HPV infections are common but difficult to distinguish. Therefore, a genotyping platform based on competitive hybridization and AMASE is described, used on clinical sample material and evaluated by comparison to Sanger DNA sequencing (Papers II and III). A flexible tag-microarray was used for detection and the two levels of discrimination gave a high level of specificity. Easy identification of multiple infections was possible which provides new opportunities to investigate the importance of multiply infected samples. To achieve highly multiplexed allele-specific extension reactions, large numbers of primers will be employed and lead to spurious hybridizations. Papers IV to VI focus on an alternative approach to control oligomerization by using protease mediated allele-specific extension (PrASE). In order to maintain stringency at higher temperatures, Proteinase K, was used instead of apyrase, leading to DNA polymerase degradation and preventing unspecific extensions. An automated assay with tag-array detection for SNP genotyping was established. First PrASE was introduced and characterized (Paper IV), then used for genotyping of 10 SNPs in 442 samples (Paper V). A 99.8 % concordance to pyrosequencing was found. PrASE is a flexible tool for association studies and the results indicate an improved assay conversion rate as compared to plain allele-specific extension. The highly polymorphic melanocortin-1 receptor gene (MC1R) is involved in melanogenesis. Twenty-one MC1R variants were genotyped with PrASE since variants in the gene have been associated to an increased risk of developing melanoma. A pilot study was performed to establish the assay (Paper VI) and subsequently a larger study was executed to investigate allele frequencies in the Swedish population (Paper VII). The case and control groups consisted of 1001 and 721 samples respectively. A two to sevenfold increased risk of developing melanoma was observed for carriers of variants. / QC 20101028

Submitted by Alessandra Portugal (alessandradf@ioc.fiocruz.br) on 2013-09-27T18:18:34Z No. of bitstreams: 1 SÉRGIO MENEZES AMARO FILHO.pdf: 3118907 bytes, checksum: 5ace372e7cf9ac4518a96c77052798ac (MD5) / Made available in DSpace on 2013-09-27T18:18:34Z (GMT). No. of bitstreams: 1 SÉRGIO MENEZES AMARO FILHO.pdf: 3118907 bytes, checksum: 5ace372e7cf9ac4518a96c77052798ac (MD5) Previous issue date: 2012-03-30 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / O câncer cervical é o terceiro tipo de câncer mais comum entre mulheres no mundo. Dentre outros cofatores, a infecção pelo HPV de alto risco, tem sido bem documentada como fator necessário ao desenvolvimento desse tipo de câncer. A principal ação do HPV envolve a expressão massiva das oncoproteínas virais E6 e E7 que podem formar complexos específicos com proteínas supressoras de tumor, sendo capazes de alterar mecanismos do ciclo celular, modificando a expressão de proteínas celulares. Um dos principais avanços na medicina clínico patológica é o uso dessas proteínas como marcadores de forma a aumentar a acurácia do prognóstico e do próprio estadiamento do câncer cervical. Assim, a fim de reforçar a hipótese de que as proteínas associadas ao ciclo celular Ki-67, MCM-2, p53 e p16INK4a se encontram superexpressas no câncer cervical, foram analisadas em 87 amostras cervicais de pacientes com câncer invasivo (CCI) e 43 cérvix normais. Verificamos também se há uma expressão diferencial que pode ajudar a avaliação do estadiamento clínico da FIGO e quais tipos virais podem induzir a uma expressão diferencial dessas proteínas. Além disso, características sociodemográficas, comportamentais e clínicas das pacientes foram obtidas dos prontuários e analisadas. Para isso, a detecção de DNA de HPV foi realizada pela técnica da PCR e hibridização in situ. A expressão das proteínas foi observada por imunohistoquímica, seguida por quantificação manual e através do software ImagePro Plus. A análise estatística foi feita utilizando o software STATA/SE 10.1 aplicando os testes: Kruskall-Wallis, Student, Fisher e Qui-Quadrado. Nossos resultados mostraram forte associação (p<0,05) do CCI e dos estágios tumorais mais avançados (III e IV) com mulheres superiores a 55 anos, com mais de quatro gestações e sem escolaridade. A prevalência de DNA de HPV por PCR na população total foi de 73,4%. Nos grupos de CCI e controle foram, respectivamente, 94,3% e 29,3%. O tipo mais prevalente foi o HPV16 (70,8%), acompanhado pelo HPV33 (11,2%) e 35 (4,5%). Como esperado, foi observado aumento (p<0,05) na expressão de Ki-67, MCM-2, p53 e p16INK4a no CCI, quando comparados ao controle. A proteína p16INK4a com expressão difusa, citoplasmática e nuclear esteve associada ao câncer. Ki-67 apresentou forte expressão (>50%) conforme o agravamento da doença. Não foi observada associação entre a expressão de MCM-2, p53 e p16, e o estadiamento do tumor. Como conclusões, foram observadas maiores chances no desenvolvimento do CCI em mulheres com idades superiores a 55 anos, com mais de quatro gestações e sem escolaridades, estando esses fatores associados, também, à progressão tumoral. Apenas o marcador Ki-67 associouse ao estágio do CCI. Os tipos mais prevalentes encontrados, HPV16, 33, 35, 67 e 58, sugerem que novos estudos devam ser considerados para implementação de vacinação contra o HPV no Brasil. / Cervical cancer is the thrid most common cancer among women worldwide. Beside others cofactors, infection with high risk HPV has been well documented as a necessary cause for cancer development. The main action involves the expression of massive HPV E6 and E7 viral oncoproteins that can form specific complexes with tumor suppressor proteins that are able of changing mechanisms of the cell cycle, modifying the expression of cellular proteins. One of the major advances in medicine clinical pathology is the use of these proteins as markers in order to improve the accuracy of prognosis and the cervical cancer stages. Thus, in order to reinforce strengthen the hypothesis that the proteins involved in cell cycle Ki-67, MCM-2, p53 and p16INK4a are over expressed in the uterine cervix of patients with cervical cancer, we analyzed 87 samples from patients with invasive cervical cancer (ICC) that were compared with 43 normal cervices (controls). Was verified also whether there is a differential expression that can help to assess the FIGO staging for ICC and which HPV viral types can induce a differential expression of these proteins. Moreover sociodemographic, behavioral and clinical characteristics of patients were obtained from medical records and analyzed. Therefore, the detection of HPV DNA was performed by PCR and in situ hybridization. By means of immunohistochemistry the expression of Ki-67, MCM2, p53 and p16 were analyzed. Statistical analysis was performed using STATA / SE 1.10 by applying the Kruskal-Wallis test, T-Student, chi-square and Fisher. Our results showed a strong correlation (p<0.05) of the CCI and the later stages of the cancer in women over 55 years, more than four pregnancies and no school education. The overall HPV DNA prevalence was 73.4%. On the ICC group and control group the prevalence was, respectively, 94.3% and 29.3%. The most prevalent HPV types were HPV16 (70.8%), HPV33 (11.2%) followed by HPV 35 (4.5%). Women with HPV16 were associated with advanced age (50.8 years) compared to women with other types (58.2 years). As expected, was observed an increase in Ki-67, MCM-2, p53 and p16 expression in CCI (p<0.05), compared to control. The expression of p16 protein with diffuse cytoplasmic and nuclear staining was associated with cancer. The Ki-67 showed a strong expression (> 50%) as the later stage of the disease. There was no association between the expression of MCM-2, p53 and p16 and tumor staging. In conclusion, we observed higher risks of ICC development in older ages, multiple pregancies and no school education. Only the Ki-67 marker was associated with the stage of the ICC. The most prevalent HPV types found, HPV 16, 33, 35, 67 and 58, suggest that new studies should be evaluated and considered on implementation of HPV vaccination in Brazil.