Functional and therapeutic significant of heat-shock protein 90 (HSP90) in reproductive cancers.

Reproductive cancers, such as ovarian, cervical, and endometrial carcinomas, have a poor prognosis in metastatic stages. Researchers are continuously seeking improved and safer methods to target cancer-related oncoproteins, addressing the limitations of current treatments, including their limited effectiveness, drug resistance, and off-target effects. Recent advancements in understanding the molecular mechanisms involved in the progress of reproductive cancers have provided valuable insights into potential targeted therapies. By engaging with oncoproteins and co-chaperones, heat-shock protein 90 (HSP90) regulates signaling networks and fixes protein folding errors in cancer cells. The potential of HSP90 inhibition as cancer-targeted treatments is underscored by the continuous discovery and testing of novel HSP90-targeted molecules for their antitumor properties in preclinical and clinical settings. Therefore, this study aims to shed light on the mechanism and recent research breakthroughs of HSP90, as well as provide an in-depth review of their therapeutic potential in reproductive cancers.

Computational insights into CRISP3 downregulation in cervical cancer and its cervical lineages pattern.

Background: Cysteine-rich secretory protein 3 (CRISP3) emerges as a potential biomarker in the study of many cancers, including cervical cancer (CC). This study aimed to analyze the expression pattern of CRISP3 in CC patients and CC cell lineages, following treatment with the epigenetic drugs: trichostatin A (TSA) and 5-aza-2'-deoxycytidine (5-aza). Methods: The differentially expressed genes identified in GSE63514 were used to construct a protein-protein interaction network. CRISP3 was selected for subsequent analyses. We utilized data from the TCGA and GENT2 projects to evaluate the expression profile and clinical behavior of CRISP3. Additionally, we conducted cell culture experiments to analyze the expression profile of CRISP3 in cells. Results: Low levels of CRISP3 were observed in squamous cell carcinoma (SCC) and human papillomavirus (HPV)16+, along with being associated with worse overall survival (OS). MIR-1229-3p was analyzed, and its high expression was associated with worse prognostic outcomes. In CC-derived cell lines, we observed low levels of CRISP3 in SiHa, followed by SW756, C33A, HeLa, and higher levels in CaSki. All cells were treated with TSA, 5-aza, or both. In all cell lines, treatment with TSA resulted in increased transcription of CRISP3. Conclusion: We identified a significant downregulation of CRISP3 in CC, particularly in cases with HPV16 infection and SCC, which was associated with poorer OS. Preliminary findings suggest that epigenetic treatments with TSA and 5-aza may modulate CRISP3 expression, warranting further research to elucidate its regulatory mechanisms and potential as a prognostic biomarker.

Vaginal Ovule Loaded with Bismuth Lipophilic Nanoparticles and Cetylpyridinium Chloride Inhibits Human Cervical Carcinoma and Candida albicans Growth.

Bismuth lipophilic nanoparticles (BisBAL NPs) and cetylpyridinium chloride (CPC) are antineoplastic and antimicrobial in vitro. As a next pre-clinical step, a clinically viable dosage form for vaginal application was developed. Compendial pharmacopeial tests (mass uniformity, disintegration, and compressive mechanics) and inductively coupled plasma optical emission spectroscopy were conducted on in-house developed glycerinated gelatin (60:15 v/w) vaginal ovules containing BisBAL NP-CPC. The antimycotic activity of BisBAL NP-CPC vaginal ovules was analyzed using disk diffusion and cell viability XTT assays. The antitumor properties of BisBAL NP-CPC vaginal ovules were assessed by cell viability MTT tests. BisBAL NP-CPC and drug-free vaginal ovules deposited into ex vivo porcine vaginas disaggregated without signs of adverse cytotoxicity within the timespan of clinical efficacy. BisBAL NP-CPC vaginal ovules demonstrated antifungal efficacy comparable to miconazole: C. albicans growth inhibition haloes in diffusion tests were 23 ± 0.968 mm (n = 3) for BisBAL NP-CPC and 20.35 ± 0.899 mm (n = 3) for miconazole. Likewise, BisBAL NP-CPC vaginal ovules reduced HeLa cell growth by 81%, outperforming the clinical reference of 500 µM 5-fluouracil, which induced a 70% growth inhibition. BisBAL NP-CPC incorporated into glycerinated gelatin vaginal ovules constitute an innovative drug delivery system for topical antimycotic and anti-cervical carcinoma treatments.

DBD-FISH Using Specific Chromosomal Region Probes for the Study of Cervical Carcinoma Progression.

Genomic instability is an important biomarker in the progression of cervical carcinoma. DBD-FISH (DNA breakage detection-fluorescence in situ hybridization) is a sensitive method that detects strand breaks, alkali-labile sites, and incomplete DNA excision repair in cells of the cervical epithelium. This technique integrates the microgel immersion of cells from a vaginal lesion scraping and the DNA unwinding treatment with the capacity of FISH integrated into digital image analysis. Cells captured within an agarose matrix are lysed and submerged in an alkaline unwinding solution that generates single-stranded DNA motifs at the ends of internal DNA strand breaks. After neutralization, the microgel is dehydrated and the cells are incubated with DNA-labeled probes. The quantity of a hybridized probe at a target sequence corresponds to the measure of the single-stranded DNA produced during the unwinding step, which is equivalent to the degree of local DNA breakage. DNA damage does not show uniformly throughout the entire DNA of a cell; rather, it is confined to specific chromosomal sites. In this chapter, an overview of the technique is supplied, focusing on its ability for assessing the association between DNA damage in specific sequences and in the progressive stages of cervical carcinoma.

Genotipos de papiloma virus en mujeres de la selva peruana con inspección visual con ácido acético positivo / Papillomavirus genotypes in women from the Peruvian jungle with a positive visual inspection with acetic acid

Objetivo: El objetivo de este estudio fue determinar la distribución de los tipos de virus de papiloma humano en mujeres con diagnóstico confirmatorio por inspección visual con ácido acético. Métodos: Se incluyeron mujeres con citología positiva para lesión intraepitelial. Los genotipos de virus de papiloma humano se analizaron mediante el sistema Xpert® HPV (GXHPV-CE-10) junto con el análisis de citología patológica de muestras de tejido cervical. Resultados: Se encontró una prevalencia de infección por virus de papiloma humano del 61 %, hubo correlación en la edad y los tipos virales detectados con el canal de color P5 (virus de papiloma humano 39, 56, 66, 68), edad de la primera relación sexual y virus de papiloma humano 18, número de gestaciones con P3 (virus de papiloma humano 31, 33, 35, 52, 58) y P5. El antecedente de haber tenido una infección de transmisión sexual se correlacionó con virus de papiloma humano 18, P3 y P4 (virus de papiloma humano 51 o 59). En la coinfección se encontró correlación entre los genotipos de virus de papiloma humano 16 con: P3 (R: - 0,11), P4 (R: - 0,22) y P5 (R: - 0,14), con haber tenido tratamiento previo (R: - 0,14). El genotipo de virus de papiloma humano 18 se correlacionó con: P3 (R: 0,28). Conclusiones: Los tipos de virus de papiloma humano de alto riesgo 16, 18 y P3 fueron los más predominantes en los grados citológicos establecidos y entre las mujeres coinfectadas(AU)

Objective: The objective of this study was to determine the distribution of papillomavirus types in women with a confirmatory diagnosis with visual inspection with acetic acid (VIA). Methods: Women with positive cytology for intraepithelial lesion were included. Papillomavirus genotypes were analyzed using the Xpert® HPV system (GXHPV-CE-10) in conjunction with pathological cytology analysis of cervical tissue samples. Results: A prevalence of human papillomavirus infection of 61% was found, there was a correlation in age and the viral types detected with the P5 color channel (human papillomavirus 39, 56, 66, 68), age of the first sexual intercourse and human papillomavirus 18, number of pregnancies with P3 (human papillomavirus 31, 33, 35, 52, 58) and P5. The history of having a sexually transmitted infection was correlated with papillomavirus 18, P3, and P4 papillomavirus 51 and 59). In coinfection, a correlation was found between papillomavirus 16 genotypes with P3 (R: - 0.11), P4 (R: 0.22), and P5 (R: - 0.14), with having had previous treatment (R: - 0.14). Papillomavirus 18 correlated with: P3 (R: 0.28). Conclusions: High-risk human papillomavirus typess 16, 18, and P3 were the most predominant in established cytological grades and among coinfected women(AU)

Tumor histology is an independent prognostic factor in locally advanced cervical carcinoma: A retrospective study.

BACKGROUND: Even with different histologic origins, squamous cell carcinoma (SCC) and adenocarcinoma (AC) are considered a single entity, and the first-line treatment is the same. Locally advanced disease at the diagnosis of cervical cancer is the most important prognostic factor, the recurrence rate is high, making it necessary to evaluate prognostic factors other than clinical or radiological staging; histology could be one of them but continues to be controversial. The aim of this study was to evaluate tumor histology as a prognostic factor in terms of treatment outcomes, disease-free survival (DFS) and overall survival (OS) in a retrospective cohort of patients with Locally Advanced Cervical Carcinoma (LACC). METHODS: The records of 1291patients with LACC were reviewed, all of them were treated with 45-50 Gy of external beam radiotherapy with concurrent chemotherapy and brachytherapy. A descriptive and comparative analysis was conducted. Treatment response was analyzed by the chi-square test; DFS and OS were calculated for each histology with the Kaplan-Meier method and compared with the log-rank test; and the Cox model was applied for the multivariate analysis. RESULTS: We included 1291 patients with LACC treated from 2005 to 2014, of which 1154 (89·4%) had SCC and 137 (10·6%) had AC. Complete response to treatment was achieved in 933 (80·8%) patients with SCC and 113 (82·5%) patients with AC. Recurrence of the disease was reported in 29·9% of SCC patients and 31·9% of AC patients. Five-year DFS was 70% for SCC and 62·2% for AC. The five-year OS rates were 74·3% and 60% for SCC and AC, respectively. The mean DFS was 48·8 months for SCC vs 46·10 for AC (p = 0·043), the mean OS was 50·8 for SCC and 47·0 for AC (p = 0·002). CONCLUSION: Our findings support the hypothesis that SCC and AC are different clinical entities. TRIAL REGISTRATION: NCT04537273 .

microRNA Profile Associated with Positive Lymph Node Metastasis in Early-Stage Cervical Cancer.

Lymph node metastasis (LNM) is an important prognostic factor in cervical cancer (CC). In early stages, the risk of LNM is approximately 3.7 to 21.7%, and the 5-year overall survival decreases from 80% to 53% when metastatic disease is identified in the lymph nodes. Few reports have analyzed the relationship between miRNA expression and the presence of LNM. The aim of this study was to identify a subset of miRNAs related to LNM in early-stage CC patients. Formalin-fixed paraffin-embedded tissue blocks were collected from patients with early-stage CC treated by radical hysterectomy with lymphadenectomy. We analyzed samples from two groups of patients-one group with LNM and the other without LNM. Global miRNA expression was identified by microarray analysis, and cluster analysis was used to determine a subset of miRNAs associated with LNM. Microarray expression profiling identified a subset of 36 differentially expressed miRNAs in the two groups (fold change (FC) ≥ 1.5 and p < 0.01). We validated the expression of seven miRNAs; miR-487b, miR-29b-2-5p, and miR-195 were underexpressed, and miR-92b-5p, miR-483-5p, miR-4534, and miR-548ac were overexpressed according to the microarray experiments. This signature exhibited prognostic value for identifying early-stage CC patients with LNM. These findings may help detect LNM that cannot be observed in imaging studies.

Evaluation of Elafin Immunohistochemical Expression as Marker of Cervical Cancer Severity.

INTRODUCTION: The main risk factor for the development of cervical cancer (CC) is persistent infection by human papillomavirus (HPV) oncogenic types. In order to persist, HPV exhibits a plethora of immune evasion mechanisms. PI3/Elafin (Peptidase Inhibitor 3) is an endogenous serine protease inhibitor involved in epithelial protection against pathogens. PI3/Elafin's role in CC is still poorly understood. MATERIALS AND METHODS: In the present study, we addressed PI3/Elafin protein detection in 123 CC samples by immunohistochemistry and mRNA expression in several datasets available at Gene Expression Omnibus and The Cancer Genome Atlas platforms. RESULTS: We observed that PI3/Elafin is consistently downregulated in CC samples when compared to normal tissue. Most of PI3/Elafin-positive samples exhibited this protein at the plasma membrane. Besides, high PI3/Elafin expression at the cellular membrane was more frequent in in situ stages I + II than in invasive cervical tumor stages III + IV. This indicates that PI3/Elafin expression is gradually lost during the CC progression. Of note, advanced stages of CC were more frequently associated with a more intense PI3/Elafin reaction in the nuclei and cytoplasm. CONCLUSION: Our results suggest that PI3/Elafin levels and subcellular localization may be used as a biomarker for CC severity.

Use of interstitial brachytherapy in pelvic recurrence of cervical carcinoma: Clinical response, survival, and toxicity.

PURPOSE: The purpose of this study was to evaluate clinical response, postrecurrence survival, disease-free survival (DFS), and toxicity related to reirradiation in pelvic recurrence of cervical carcinoma. METHODS AND MATERIALS: A retrospective cohort study of 45 women undergoing high-dose-rate interstitial brachytherapy (HDR-IB) was conducted from 1998 to 2014. Clinical information, as well as data on the malignancy, primary treatment, HDR-IB technique, and toxicity, was collected. Statistical analysis used chi-square or Fisher's exact test, Kaplan-Meier survival curves and log-rank test, and Cox regression, with p < 0.05 for significance. RESULTS: There were 30 cases (67%) of complete clinical response, with a followup period of 9-129 months (20 alive, 10 died). The 5-year postrecurrence survival rate was 52%. Among 15 women without complete clinical response, the survival rate was low (<8 months). In the 30 women with complete clinical response, the 5-year DFS was 42%. All analyzed variables were not associated with survival. Ultrasonography-based needle placement was not associated with disease control or toxicity. Toxicity was reported in 23 women (51%) with 14 fistulas, unrelated to clinical response. However, there was a higher occurrence of fistula when chemotherapy was used. CONCLUSIONS: Reirradiation using HDR-IB for pelvic recurrence of cervical carcinoma yielded a good complete clinical response rate. Postrecurrence survival and DFS rates were higher than expected, equivalent to salvage surgery, but with significant toxicity. Despite toxicity, this technique can be an alternative for selected cases.

Biomarkers of resistance to radiation therapy: a prospective study in cervical carcinoma.

BACKGROUND: Clinical parameters and proteins have recently been suggested as possible causes of radiotherapy (RT) resistance in cervical carcinoma (CC). The objective of the present study was to validate prognostic biomarkers of radiation resistance. METHODS: The present prospective study included patients undergoing RT with curative intent for histologically proven locally advanced squamous cell CC. Tissues and blood samples were systematically collected before RT initiation. Immuno-histochemistry was performed (IGF-IR α and ß, GAPDH, HIF-1 alpha, Survivin, GLUT1, CAIX, hTERT and HKII). Response to radiation was assessed through tumour response 3 months after RT completion, through overall survival (OS) and through progression-free survival (PFS). RESULTS: One hundred forty nine patients with a mean age of 46 years were included, with FIGO IIB (n = 53) and FIGO IIIB (n = 96) CCs. 61 patients were treated with exclusive RT + brachytherapy and 88 underwent chemo-radiotherapy + brachytherapy. Our findings suggest an association between hemoglobin level (Hb) (>11 g/dL) and 3 months complete response (p = 0.02). Hb level < 11 g/dL was associated with decreased PFS (p = 0.05) and OS (p = 0.08). Overexpression of IGF-1R ß was correlated with a decreased OS (p = 0.007). Overexpression of GLUT1 was marginally correlated with reduced OS (p = 0.05). PFS and OS were significantly improved in patients undergoing chemoradiation versus exclusive radiotherapy (PFS: p = 0.04; OS: p = 0.01). CONCLUSIONS: IGF-1R ß overexpression and Hb level (≤11 g/dl) were associated with poor prognosis, and thus appear to be possible interesting biomarkers of radiation resistance. Our results corroborate previous pre-clinical studies suggesting IGF-1R and hypoxia to be part of the biological pathways leading to radio-resistance.

HPV genotype distribution and anomalous association of HPV33 to cervical neoplastic lesions in San Luis Potosí, Mexico.

BACKGROUND: The association of human papillomavirus (HPV) types to neoplastic lesions increase as a function of their oncogenicity and the duration of the infection since lesion severity progresses from low-grade to high-grade and cancer. In an outbreak, the prevalence of the HPV type involved would increase and the proportion of the associated low-grade lesions would predominate over severe lesions. In this study, the prevalence of HPV types and their association to neoplastic lesions was determined in women subjected to colposcopy in San Luis Potosí, Mexico. METHODS: DNA from high-risk (HR) and low-risk (LR) HPV types was identified by E6 nested multiplex PCR in cervical scrapes from 700 women with normal cytology, atypical squamous cells of undetermined significance (ASCUS), low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL) or invasive cervical cancer (CC). RESULTS: Overall HPV-DNA prevalence was 67.7 %, that of HR-HPV was 63.1 %, and that of LR-HPV was 21.3 %. The highest prevalence (78.2 %) occurred in the 15-24 year group, whereas that of single infections was 52 % and that of multiple infections (i.e., by 2-6 HPV types) was 48 %. The most prevalent HR types were HPV33 (33.1 %), HPV16 (16.6 %), HPV18 and HPV51 (6.7 % each). HR-HPV prevalence was 29.6 % in normal cytology, 26.7 % in ASCUS, 63.3 % in LSIL, 68.2 % in HSIL, and 90.5 % in CC. Three prevalence trends for HR-HPV types were found in neoplastic lesions of increasing severity: increasing (LSIL < HSIL < CC) for HPV16, HPV39, HPV18, HPV58, HPV31 and HPV35; asymptotic (LSIL < HSIL ≈ CC) for HPV51 and HPV68; U-shaped (LSIL < HSIL > CC) for HPV33. CONCLUSIONS: Two-thirds of the women subjected to colposcopy from 2007 to 2010 in San Luis Potosí have HPV infections which predominate in the 15-24 years group. Around half of the infections are by one viral type and the rest by 2-6 types. HPV33 is the most prevalent type, followed by HPV16. Overall HR-HPV prevalence increases with the severity of neoplastic lesions. HPV33 prevalence is highest in LSIL and its U-shaped trend with progressing neoplastic lesions differs from the growing/asymptotic trends of other HR-HPV types. An ongoing or recent HPV33 outbreak is consistent with its high prevalence and anomalous association to LSIL.

Concurrent radiotherapy with S-1 plus cisplatin versus concurrent radiotherapy with cisplatin alone for the treatment of locally advanced cervical carcinoma: a pilot randomised controlled trial.

BACKGROUND: In the present study, we compared the efficacy and safety of concurrent radiotherapy with S-1 plus cisplatin (CRSC) versus concurrent radiotherapy with cisplatin alone (CRC) for the treatment of advanced cervical carcinoma (ACC). METHODS: Between February 2006 and January 2009, 72 eligible patients with ACC were included and randomly divided into two groups. Thirty-six patients received CRSC with radiotherapy (60 Gy/30 fractions over 6 weeks) beginning on day 1, S-1 (according to body surface area) for 28 days repeated every 6 weeks, and cisplatin (50 mg/m(2), intravenously on day 1) every 4 weeks for two cycles. The other 36 received CRC at the same cisplatin and radiotherapy dosage as for CRSC. The primary outcome was overall survival, whereas the secondary outcomes included progression-free survival and toxicity. RESULTS: The median overall survival was 75 months (range 4-86 months) for the CRSC group and 66 months (range 3-87 months) for the CRC group (P = 0.039). The median corresponding progression-free survival was 66 months (range 3-75 months) and 58 months (range 3-71 months), respectively (P = 0.042). The toxicity profile was similar in both the groups. CONCLUSION: Our results suggested that CRSC might be more effective than CRC in patients with ACC with acceptable toxicity.

Histerectomía radical videolaparoscópica / Radical Hysterectomy Video-laparoscopy

Introducción: desde el primer intento de realizar una histerectomía radical laparoscópica 20 años atrás, los estudios comparativos de casos control han demostrado que el procedimiento laparoscópico consume más tiempo (30-60 min más que el convencional), pero la pérdida de sangre es menor, el movimiento intestinal se recupera rápidamente y se reduce la hospitalización con resultados posoperatorios similares.Objetivo: demostrar la factibilidad y seguridad del procedimiento en el tratamiento del cáncer de cuello uterino en estadios iniciales.Métodos: se realizó un estudio observacional descriptivo de una serie consecutiva de 15 pacientes a las que se les practicó una histerectomía radical laparoscópica en el Centro Nacional de Cirugía de Mínimo Acceso en el periodo comprendido entre febrero de 2012 y julio de 2013. Las variables estudiadas fueron: edad, raza, variedad histológica, tiempo quirúrgico, sangrado transoperatorio, permeación vascular y linfática, afectación ganglionar, tratamiento adyuvante y complicaciones posoperatorias. Se emplearon porcentajes, promedios y desviación estándar como medidas de resumen de las variables.Resultados: la edad media de las pacientes fue de 42,8±10 años con un rango entre 28 y 60 años. El tiempo quirúrgico fue de 235,3±39,3 min. El sangrado medio fue de 98,3 mL±42,3 mL. No se presentaron complicaciones transoperatorias. El número medio de ganglios extirpados fue de 17,7, y un caso único con metástasis ganglionar (6,7 por ciento). Dos pacientes recibieron tratamiento adyuvante (13,3 por ciento).Conclusiones: la histerectomía radical videolaparoscópica constituye un procedimiento factible y seguro para el tratamiento del cáncer de cuello uterino en etapas tempranas (Ib1-IIa1)(AU)

Cervical cytology, HPV DNA and mRNA - a comparative study in 162 patients / Citologia cervicovaginal, DNA HPV e RNAm - estudo comparativo em 162 pacientes

In contrast to the general improvement of the socioeconomic status of the Brazilian population, pathologies that are characteristic of poor health assistance persist. Among those, cervical cancer (CC) is emblematic; it still presents a persistently high incidence. Objective: to compare the performance of cervical cytology to HPV DNA and mRNA detection methods in 162 patients undergoing routine gynecological clinical practice. Methods:a total of 162 patients attended during routine gynecological examination in a private clinic in Florianópolis, Santa Catarina, Brazil, had cervical samplescollected and processed for cytopathological and molecular tests, conventional PCR and NASBA. Positive samples positive for HPV DNA were submittedto Type-Specific PCR (TS-HPV PCR). Patients with altered smears were submitted to colposcopy and biopsy. Results: among the 162 samples, 19.8%(32/162) had altered smears, being 4/32 classified as ASC-H, 9/32 as ASC-US, 9/32 as LSIL and 10/32 as HSIL. Biopsies revealed nine cases of CIN I,nine CIN II and one CIN III, while seven were negative for cervical neoplasia. Overall, HPV DNA was detected in 38.3% (62/162) of the samples and HPV E6/E7 mRNA expression was found in 13.6% (22/162). Using TS-HPV PCR, HPV 16 was the most frequent type, found in 8% of the samples (5/62).Considering CIN2+ the gold-standard, cytology had 38.5% of specificity. Sensitivity and specificity of HPV-DNA PCR and NASBA were, respectively,100% and 60%; 18.7% and 68.7%. Conclusion: mRNA E6/E7 expression was not a highly specific or sensitive marker for prevalent cervical disease while HPV DNA may be used for cervical cancer screening only in conjunction to more specific adjuvant tests.

Em contraste com a melhora geral da situação socioeconômica da população brasileira, patologias que são características de uma deficiente assistência à saúde persistem. Entre elas, o câncer cervical (CC) é emblemático, ainda apresentando uma persistente alta incidência. Objetivo: avaliaro desempenho da citologia e de métodos de detecção de DNA e RNAm de HPV em 162 pacientes submetidas a prática clínica ginecológica de rotina.Métodos: cento e sessenta e duas pacientes atendidas em uma clínica particular de Florianópolis, Santa Catarina, Brasil, tiveram amostras cervicais coletadas e processadas para estudo citopatológico e molecular; PCR convencional e NASBA. Amostras positivas para o DNA do HPV foram submetidas àPCR tipo-específica (PCR HPV-TE). Resultados: entre as 162 amostras, 19,8% (32/162) apresentaram esfregaços alterados, sendo 4/32 classificadas comoASC-H, 9/32 como ASC-US, 9/32 como LSIL e 10/32 como HSIL. Biópsias revelaram nove casos de NIC I, nove casos de NIC II e um caso de NIC III. ODNA do HPV foi detectado em 38,3% (62/162) das amostras. Expressão de E6/E7 (RNAm) foi encontrada em 13,6% (22/162) das amostras. Utilizando a PCR tipo-específica (HPV-TE), o HPV 16 foi o tipo mais frequente, encontrado em 8% (5/62) das amostras HPV+. Considerando NIC 2+ o padrão-ouro,a especificidade da citologia foi de apenas 38,5%, enquanto a sensibilidade e a especificidade da PCR DNA e RNAm foram, respectivamente, 100% e 60%;18,7% e 68,7%. Conclusão: a expressão de E6/E7 RNAm não se mostrou um marcador altamente específico ou sensível para doença cervical prevalente,enquanto o DNA HPV pode ser utilizado para rastreamento apenas em conjunto com testes adjuvantes mais específicos.

Co-infections associated with human immunodeficiency virus type 1 in pregnant women from southern Brazil: high rate of intraepithelial cervical lesions

Human immunodeficiency virus type 1 (HIV-positive) pregnant women require specific prophylactic and therapeutic approaches. The efficacy of established approaches is further challenged by co-infection with other sexually transmitted diseases (STDs). The objective of this study was to determine the prevalence of co-infections in pregnant women infected with different HIV-1 subtypes and to relate these findings, together with additional demographic and clinical parameters, to maternal and infant outcomes. Blood samples from pregnant women were collected and tested for syphilis, hepatitis B virus (HBV) and hepatitis C virus (HCV). Human papillomavirus (HPV) diagnosis was evaluated by the presence of alterations in the cervical epithelium detected through a cytopathological exam. Medical charts provided patient data for the mothers and children. Statistical analyses were conducted with STATA 9.0. We found a prevalence of 10.8 percent for HCV, 2.3 percent for chronic HBV, 3.1 percent for syphilis and 40.8 percent for HPV. Of those co-infected with HPV, 52.9 percent presented high-grade intraepithelial lesions or in situ carcinoma. Prematurity, birth weight, Apgar 1' and 5' and Capurro scores were similar between co-infected and non-co-infected women. The presence of other STDs did not impact maternal and concept outcomes. More than half of the patients presenting cervical cytology abnormalities suggestive of HPV had high-grade squamous intraepithelial lesions or cervical cancer, evidencing an alarming rate of these lesions.

Radical vaginal trachelectomy and laparoscopic pelvic lymphadenectomy in IB1 cervical cancer during pregnancy.

► We expose an novel surgical technique in pregnant women with cervical cancer. ► We preserve the pregnancy in this case. ► After 40 months of monitoring both patient and her child are healthy.

Histerectomía extrafascial en cáncer de cuello uterino IB2 posterior a radio-quimioterapia / Extrafascial hysterectomy in IB2 cervical cancer after radiochemotherapy

Objetivo: Evaluar la sobrevida global y libre de enfermedad en pacientes con tratamiento concomitante de radioquimioterapia más histerectomía extrafascial en cáncer de cuello uterino IB2. Método: 31 pacientes con cáncer cérvico-uterino en etapa IB2 fueron tratadas con radioquimioterapia concomitante más histerectomía extrafascial programada en 4 a 6 semanas. Se efectuó radioterapia pelviana externa e implante de braquiterapia útero-vaginal con dosis preoperatoria entre 70-75 Gy a punto A y quimioterapia con cisplatino. Resultados: La edad promedio de las pacientes fue de 41 +/- 8 años. El 81 por ciento de los casos fue carcinoma escamocelular. La histerectomía extrafascial se realizó 4 a 6 semanas post-radiación en el 85 por ciento de los casos. El 79 por ciento y el 13 por ciento de las pacientes recibieron 5 y 4 ciclos de quimioterapia respectivamente. La mediana de seguimiento fue de 38 meses. La sobrevida global y libre de enfermedad estimada a 5 años fue de 86 por ciento y 79 por ciento respectivamente. Hubo diferencia significativa en la sobrevida global en los subgrupos sin o con enfermedad residual macroscópica en la pieza operatoria (p<0,001). Conclusión: Esta serie muestra resultados de sobrevida similares a las reportadas en otros trabajos con el mismo tratamiento. La enfermedad residual en la pieza operatoria podría constituir un factor de valor pronóstico.

Aim: To evaluate overall and disease free survival in cervical cáncer IB2 patients with concomitant chemo-radiotherapy and posterior extrafascial hysterectomy treatment. Methods: Between July 2005 to December 2009, a total of 31 eligible patients with IB2 cervical cancer were treated with chemoradiotherapy and posterior hysterectomy in the Carlos Van Buren Hospital Oncology Unit. Radiotherapy consisted in external pelvic radiation and a utero-vaginal brachitherapy with low rate doses to get preoperatory doses between 70 and 75 Gy to the A point and a concomitant Cisplatin based chemotherapy. After this treatment, the patients received extrafascial hysterectomy 4 to 6 weeks completed chemoradiotherapy. Results: The mean age was 41 +/- 8 years. 81 percent of the patients had an spinocelular carcinoma. The extrafascial hysterectomy was made between 4 to 6 weeks post-radiation in 85 percent of the patients. 79 percent and 13 percent of the patients received 5 and 4 chemotherapy cycles respectively. The median follow up was 38 months. The 5 years overall survival and disease free survival estimates were 86 percent and 79 percent respectively. There was a significant difference between subgroups of patients with and without macroscopic residual disease in the operatory specimen (p<0.001). Conclusion: Our survival is similar to published results with the same treatment. The presence of macroscopic residual disease in the hysterectomy specimen could be a factor of prognostic value.

Radioterapia lumboaórtica en pacientes con cáncer de cuello uterino: experiencia del Instituto Nacional del Cáncer / Paraortic radiotherapy in patients with uterine cervical carcinoma: Instituto Nacional del Cáncer experience

Antecedentes: El cáncer de cérvicouterino continúa siendo una enfermedad prevalente en Chile. Es frecuente recibir pacientes en etapas IIB y IIIB donde el riesgo de compromiso ganglionar pelviano y lumbo-aórtico es elevado y el tratamiento es radioquimioterapia. Objetivo: Análisis retrospectivo de pacientes con cáncer cérvicouterino que recibieron radioterapia que incluía los territorios lumboaórticos. Método: Revisión de pacientes con cáncer cérvicouterino tratadas entre 1995 y 2007 en el Instituto Nacional del Cáncer, Santiago, Chile. En 39 pacientes el tratamiento incluyó las cadenas lumboaórticas. Se analizó toxicidad y evolución. Se utilizaron dos técnicas de radioterapia: la primera durante la década del 90, donde se empleaban dos campos paralelos y opuestos (anterior y posterior) y una segunda actual, donde se irradian en conjunto la pelvis y los lumboaórticos a través de 4 campos (laterales y AP-PA). Resultados: El análisis dosimétrico de las dos técnicas confirma un mayor volumen irradiado de tejidos normales con la técnica de dos campos, fundamentalmente intestino delgado. La toxicidad fue significativamente diferente, siendo la que utilizamos hoy menos tóxica y con bajas complicaciones gastrointestinales, a pesar de recibir un tratamiento de radio-quimioterapia concomitante (55 por ciento vs 0 por ciento). Conclusión: La radioterapia lumboaórtica es un tratamiento con una tolerancia aceptable incluso con quimioterapia concomitante. Esta revisión nos obliga a elaborar un nuevo trabajo para evaluar si la radioterapia lumboaórtica reduce el riesgo de recidiva y aumenta la sobrevida.

Background: Uterine cancer is still a prevalent disease in Chile. Is common to treat patients with tumors in stages IIB and IIIB where the risk of pelvic and paraortic limph node involvement is very high. Its treatment is radio-chemotherapy. Objective: To present a retrospective analysis of patients that suffered cervix-uterine cancer who were treated with radiotherapy including the aortic-lumbar area. Methods: From the revision of patients who were treated of cervix-uterine cancer between the years 1995 and 2007, 39 were treated including aortic-lumbar chains. Evolution and toxicity were analyzed. Two radiotherapy techniques were used. The first one, during the nineties, included two parallel previous and later and opposed fields, and a second technique, currently used, where pelvis and paraortic are radiated at the same time through four lateral (AP-PA) fields. Results: The dosimeter analysis of both techniques shows that there is a higher volume of radiated normal tissue with the two fields techniques, mainly in the small bowel. On the other hand, the toxicity was significantly different being today's technique less toxic and showing low gastrointestinal side effects, despite receiving a simultaneous radio-chemotherapy (55 percent vs 0 percent). Conclusion: The paraortic radiotherapy treatment has an acceptable level of tolerance even under simultaneous radio-chemotherapy. Given the results of this study, we see the need for undergoing a new research project in order to evaluate if the aortic-lumbar radiotherapy may reduce the risk of relapse and increase in survival rate.

Caracterização de metaloproteinases de matriz e reck em queratinócitos primários que expressam oncoproteínas do papilomavírus humano (HPV) / Characterization of matrix metalloproteinases and reck in primary keratinocytes that express human papillomavirus (HPV) oncoproteins

Os tumores da cérvice-uterina, que representam uma das principais doenças ginecológicas em mulheres na idade reprodutiva em todo o mundo, estão etiologicamente associados com a infecção pelo papilomavírus humano (HPV). A progressão de uma lesão intraepitelial escamosa de baixo-grau (LSIL) a um carcinoma invasivo de cérvix uterina está acompanhada da degradação da matriz extracelular (MEC) devido à ação progressiva das metaloproteinases de matriz (MMP-2, MMP-9 e MMP-14) no processo de invasão e metástase. Entretanto, o balanço entre as MMPs e seus reguladores como RECK e TIMPs é necessário para controlar esta invasão. O objetivo deste projeto consiste em avaliar a atividade e a expressão das metaloproteinases 2, 9, e 14, e caracterizar a expressão do gene supressor de metástase RECK e do inibidor tecidual de metaloproteinases (TIMP-2), em modelo de queratinócitos humanos infectados com retrovírus recombinantes que expressam os oncogenes E6 e/ou E7 de HPV 16, em culturas cultivadas em monocamada e organotípicas. Para isso, utilizamos ensaios de real-time PCR, zimografia, western blot, imunocitoquímica, ensaio de ELISA e imunohistoquímica. Em culturas em monocamada observamos que as células que expressam as oncoproteínas E6E7 de HPV16 apresentaram menores níveis protéicos de RECK e TIMP-2 em relação ao controle pXLSN. Quando analisamos as culturas organotípicas, também observamos esta diminuição dos níveis de RNAm e protéicos de RECK em rafts que expressam E6E7, acompanhado pelo aumento da atividade de MMP-9, em relação ao controle. Também observamos que o tratamento das culturas com a citocina TNF aumenta a expressão gênica, protéica e atividade de MMP-9 em todas as linhagens analisadas. Além disso, os oncogenes E6 e/ou E7 não afetam a expressão e/ou atividade de MMP-2, MT1-MMP. Nossos dados demonstraram que a expressão das oncoproteínas E6E7 de HPV16 estão relacionadas com o desequilíbrio entre MMPS e seus inibidores, sugerindo que em uma fase pré-invasiva do...

Cervical cancer is etiologically associated with to high-risk human papillomavirus (HPV) infection. It has been observed that matrix metalloproteinases (MMPs) -2, -9, and MT1-MMP are required for basement membrane degradation during cervical carcinoma progression. Moreover, a counterbalancing among MMPs and their regulators, such as TIMPs and RECK, is necessary to modulate invasion. In order to study the effect of HPV oncogenes on MMPs expression, primary human keratinocytes (PHKs) were infected with recombinant retroviruses expressing wild-type HPV16 E6 and/or E7 oncogenes and were used to seed monolayers and organotypic cultures. Quantitative real-time PCR (Q-PCR), western blot, zimography, immunocitochemistry, ELISA assay and immunohistochemistry were used to determine the expression level and activity of MMP-2, MMP-9, MT1-MMP and their inhibitors RECK and TIMP-2. We observed that cultures expressing E6E7 presented lower RECK and TIMP-2 protein levels than control keratinocytes. In addition, rafts cultures presented the same lower RECK levels additionally presenting higher MMP-9 activity than control. Furthermore, we observed that expression of E6 and/or E7 proteins do not affect MMP-2 and MT1-MMP protein levels and/or activity. We also observed that TNF treatment enhance the MMP-9 gene and protein expression and activity in all studied cell lines. Taken together, our results demonstrate that HPV16E6E7 expression is related with the unbalance between MMPs and their inhibitors, suggesting that in the initial steps of HPV-related cervical disease, not only MMPs but also RECK and TIMP-2 are critical for tumor progression.