Julisa Chamorro Lascasas Ribalta

Gene network reconstruction reveals cell cycle and antiviral genes as major drivers of cervical cancer

Although human papillomavirus (HPV) was identified as an etiological factor in cervical cancer, the key human gene drivers of this disease remain unknown. Here we apply an unbiased approach integrating gene expression and chromosomal aberration data. In an independent group of patients, we reconstruct and validate a gene regulatory meta-network, and identify cell cycle and antiviral genes that constitute two major sub-networks up-regulated in tumour samples. These genes are located within the same regions as chromosomal amplifications, most frequently on 3q. We propose a model in which selected chromosomal gains drive activation of antiviral genes contributing to episomal virus elimination, which synergizes with cell cycle dysregulation. These findings may help to explain the paradox of episomal HPV decline in women with invasive cancer who were previously unable to clear the virus.

New approach reveals CD28 and IFNG gene interaction in the susceptibility to cervical cancer

Cervical cancer is a complex disease with multiple environmental and genetic determinants. In this study, we sought an association between polymorphisms in immune response genes and cervical cancer using both single-locus and multi-locus analysis approaches. A total of 14 single nucleotide polymorphisms (SNPs) distributed in CD28, CTLA4, ICOS, PDCD1, FAS, TNFA, IL6, IFNG, TGFB1 and IL10 genes were determined in patients and healthy individuals from three independent case/control sets. The first two sets comprised White individuals (one group with 82 cases and 85 controls, the other with 83 cases and 85 controls) and the third was constituted by non-white individuals (64 cases and 75 controls). The multi-locus analysis revealed higher frequencies in cancer patients of three three-genotype combinations [CD28+17(TT)/IFNG+874(AA)/TNFA-308(GG), CD28+17(TT)/IFN+847(AA)/PDCD1+7785(CT), and CD28 +17(TT)/IFNG+874(AA)/ICOS+1564(TT)] (P < 0.01, Monte Carlo simulation). We hypothesized that this two-genotype [CD28(TT) and IFNG(AA)] combination could have a major contribution to the observed association. To address this question, we analyzed the frequency of the CD28(TT), IFNG(AA) genotype combination in the three groups combined, and observed its increase in patients (P = 0.0011 by Fishers exact test). The contribution of a third polymorphism did not reach statistical significance (P = 0.1). Further analysis suggested that gene-gene interaction between CD28 and IFNG might contribute to susceptibility to cervical cancer. Our results showed an epistatic effect between CD28 and IFNG genes in susceptibility to cervical cancer, a finding that might be relevant for a better understanding of the disease pathogenesis. In addition, the novel analytical approach herein proposed might be useful for increasing the statistical power of future genome-wide multi-locus studies.

Survivin and telomerase expression in the uterine cervix of women with human papillomavirus-induced lesions.

Introduction: Infection by human papillomavirus is the most important risk factor in the pathogenesis of uterine cervical cancer. The aims of this study were to evaluate the expression of survivin protein and telomerase enzyme in samples of uterine cervix from women with human papillomavirus-induced lesions and to determine the relationship between survivin and telomerase expression and the different grades of cervical squamous intraepithelial neoplasia and invasive cervical carcinoma. Methods: Biopsy samples from the uterine cervix of 105 women aged 18 to 80 years were analyzed. The patients were divided into 5 groups: WN group, 20 patients without neoplasia; CIN-1 group, 24 patients with grade 1 cervical intraepithelial neoplasia (CIN), grade 1; CIN-2 group, 20 patients with CIN grade 2; CIN-3 group, 24 patients with CIN, grade 3; and ICC group, 17 patients with invasive cervical carcinoma. Human papillomavirus detection, telomerase activity, and survivin expression were assessed using polymerase chain reaction (PCR), real-time PCR (RT-PCR), and immunochemistry, respectively. Results: There was a significant increase in the expression of telomerase and survivin associated with the severity of the lesion. Conclusions: The results suggest that mechanisms that promote both cell proliferation (telomerase activity) and cell survival (survivin expression) are active in cervical cancer and its precursor lesions. There was a negative correlation between survivin expression and the number of PCR cycles necessary to detect telomerase activity in the total sample, achieving statistical significance in patients in the CIN-3 group.

Relação entre diagnóstico citopatológico de neoplasia intra-epitelial cervical e índices de células CD4+ e de carga viral em pacientes HIV-soropositivas

PURPOSE: to correlate the type of cervical lesion diagnosed by Pap smear with CD4 cell counts and HIV-RNA viral load in HIV-positive patients. METHODS: one hundred and fifteen HIV patients were evaluated retrospectively in the present study, during the period from January 2002 to April 2003, at a university hospital. Eighty-three patients presented cervical intraepithelial neoplasia (CIN) in Pap smear, in comparison with thirty-two with no lesions. Patients were divided into three groups, according to CD4 counts: CD4 more than 500 cells/mm3, between 200 and 500 cells/mm3, and less than 200 cells/mm3, and other three groups, according to HIV viral load: less than 10,000 HIV-RNA copies/mL, between 10,000 and 100,000 HIV-RNA copies/mL, or more than 100,000 HIV-RNA copies/mL. Correlation was investigated by the Fisher test. RESULTS: of the eighty-three patients with CIN, 73% presented CD4 counts less than 500 cells/mm3. In all CD4 groups, more than 50% of the patients presented CIN. According to the viral load, 71.7% of the patients with less than 10,000 HIV-RNA copies/mL presented CIN I, compared with 11.3% that showed CIN III. In the group with higher viral load (>100.000 HIV-RNA copies/mL), 61.5% showed CIN I and 30.8% presented CIN III. CONCLUSION: association between viral load and CIN was established (p=0.013), which was not observed with CD4 cell counts and CIN. Concomitant cervicovaginal infection was considered a potential confounding factor.

Avaliação dos Métodos Empregados no Programa Nacional de Combate ao Câncer do Colo Uterino do Ministério da Saúde

Purpose: to evaluate a populational sample of the screening proposed by the National Program of Uterine Cervical Cancer Control (PNCC), regarding the following issues: frequency of unsatisfactory cytologic results, cytologic frequency of atypical squamous or glandular cells of undetermined significance (ASCUS, AGCUS), low- or high-grade squamous intraepithelial lesions (SIL), comparing the cytologic results with anatomicopathological results of colposcopically directed biopsies. Methods: through the written, broadcasting television and oral midia, women between 35 and 49 years were requested to have a preventive cytopathological test, to be collected by the authorized public health or other institutions accredited by SUS. The slides were analyzed by the program-authorized laboratories and all those patients from the populational sample from the municipality of Navirai in the State of Mato Grosso do Sul with cellular alterations were submitted to colposcopy and directed biopsy. Results: the frequency of cytologic alterations of the ASCUS, AGCUS and SIL types was 3.3%, an index that is close to that predicted by the PNCC (4%); the percentage of samples that were unsatisfactory for evaluation was high (12.5%); among the ASCUS, AGCUS or low grade-SIL patients, 27.3% presented intraepithelial lesions of a high grade in the anatomicopathological study; while patients with cytology compatible with high grade-SIL, the directed biopsy revealed that 12.5% presented low-grade intraepithelial lesions. Conclusions: the choice of oncological cytology as the only method for the screening in the program allowed high indexes of false-negatives (27.3%) and of false-positives (12.5%). In the screening of cervical neoplasms, colposcopy has shown to be an important and indispensable method to guide the therapeutical management to be adopted.

Angiogênese na Neoplasia Escamosa do Colo Uterino: Comparação entre dois Marcadores de Células Endoteliais

Purpose: to compare the efficiency of anti-factor VIII and anti-CD34 antibodies as vascular makers in cervical cancer, in cervical intraepithelial neoplasia and in normal cervix. Methods: using an immunohistochemical method, factor VIII-related antigen and leukocyte antigen CD34, we performed microvascular counts in 18 squamous cell carcinomas, in 15 cervical high-grade intraepithelial neoplasia, in 15 low-grade intraepithelial lesions and in 10 normal cervices. Using light microscopy we counted microvessels per 400X field in the most active areas of neovascularization with higher microvessel density in each case. Results: the average of microvessels stained with anti-CD34 in invasive carcinoma, high-grade intraepithelial lesions, low-grade intraepithelial lesions and in the normal cervices was 154, 134, 112 and 93, respectively. When we used anti-factor VIII the average was 56, 44, 33 and 30 vessels, following the same order. High-grade intraepithelial lesions and invasive carcinomas showed greater means number of vessels than normal tissue. Conclusions: the use of anti-CD34 allowed the detection of a greater number of vessels when compared to anti-factor VIII. However, we could observe that anti-factor VIII staining was able to significantly discriminate high-grade from low-grade lesions.

High Levels of Granzyme B Expression in Invasive Cervical Carcinoma Correlates to Poor Response to Treatment

The present study assessed, in cervical carcinoma, expression levels of seven immune response genes and sought correlation to response to treatment. The expression levels of CD28, CTLA4, ICOS, ICOSL, CD80 and CD86 and granzyme B genes were assessed by real-time RT-PCR in pre-treatment tumor fragments. During the six-month follow-up after treatment, 8 patients presented tumor and 10 survived free of tumor. The only gene whose expression levels were higher in patients with poor outcome (p = 0.03) was granzyme B. Further evaluation, in adequately powered prospective studies is warranted to confirm the data and to translate this observation to the clinical setting.

Fibroma de Vulva (Molluscum Pendulum): Relato de Caso

A vulvar fibroma, of the molluscum pendulum type, was present in a 20-year-old patient. The tumor began to develop slowly after her menarche, when she was 14 years of age. The physical examination revealed a mass with considerable volume, painless, located at the upper third of the greator left lip, elastic consistency, greater diameter at its distal portion measuring 12 cm by 23 cm in length. The treatment was exeresis from the base of the pedicle, under local anesthesia. The tumor weighed 950 g. A literature review is included.

Alteraçöes na expressäo do antígeno nuclear de proliferaçäo celular e dos receptores de estrogênio e de progesterona provocadas pela quimioterapia primária no carcinoma de mama

ABSTRACT PURPOSE: to evaluate the changes in the cell phenotype determined by primary chemotherapy. METHODS: we evaluated the expression of proliferating cells of nuclear antigen (PCNA) and the estrogen (RE) and progesterone (RP) receptors in 17 stage II breast cancer patients before and after chemotherapy by immunohistochemistry. The values were compared with menopausal status, tumoral clinical response and with axillary lymph node status. RESULTS: there was a significant decrease in the average index of anti-PCNA-stained cells before (time A) and after (time B) chemotherapy (p=0.041). Responder patients displayed a significant decrease in PCNA levels [time A=53.1 and time B= 30.7 (p=0.011)]. A similar trend was observed in patients with histologic grade GII/GIII [time A=63.1 and time B=38.7 (p=0.049)]. There was no significant difference in PCNA expression regarding menopause status and axillary lymph node involvement. There was a significant decrease in RE after chemotherapy in the premenopausal patients [time A=60.3 and time B=24.1 (p=0.027)] and in those who showed a therapeutic response [time A=59.1 and time B=37.9 (p=0.030)]. We observed a significant increase in RP after chemotherapy in the postmenopausal patients [time A=35.3 and time B=58.3 (p=0.023)]. There was no relationship between hormone receptors and axillary lymph nodes. CONCLUSIONS: the decrease in PCNA levels in patients with high histologic grade, in RE in premenopausal patients, and both, PCNA and RE, in the tumors with clinical response after chemotherapy shows that the drugs acted on proliferating cells, and therefore PCNA can be used as a parameter of treatment response.

Cervical cancer screening in young and elderly women of the Xingu Indigenous Park: evaluation of the recommended screening age group in Brazil

Objective To analyze the occurrence of atypia in the cytology/histology examinations of young women under the age of 25 years and of elderly women aged over 64 years, in the Xingu Indigenous Park and to evaluate, in a subjective manner, if the age range for screening established by the Ministry of Health and the Instituto Nacional de Câncer is appropriate for this population. Methods The Xingu/UNIFESP Project, in partnership with the Center for Gynecological Disease Prevention, develops programs to prevent cervical cancer. The exploratory, retrospective and descriptive study of cytological and histopathological examinations of young (12-24 years) and elderly (aged 64 and over) women of the Xingu Indigenous Park, between 2005 and 2011. Results There was low occurrence of cytological atypia in the elderly female population, but there were occasional high-grade lesions in the indigenous youth. Conclusion Interrupting screening at the limit age of 64 years, as established by the Ministry of Health and the Instituto Nacional de Câncer is justified. However, screening of young women should begin at an earlier age.