Cláudio Galleano Zettler

Evidence for an association of human papillomavirus and breast carcinomas

Human papillomavirus (HPV) DNA has been detected in breast carcinoma by different laboratorial techniques, suggesting the virus could play a role in the pathogenesis of this tumor. The aim of the present study is to investigate the presence of HPV in patients with breast carcinoma and the correlation of the viral infection with prognostic factors for the disease outcome. Between June 2001 and July 2002, 101 paraffin embedded breast carcinoma specimens were analyzed through polymerase chain reaction (PCR) and sequencing of HPV-E6 gene. Twenty specimens of reduction mammoplasty and 21 specimens of fibroadenomas were also studied as a non-malignant control group. Two different specific primer sets targeting E6 region of the HPVs 16 and 18 were used for the analysis. The HPV DNA was detected in 25 breast carcinomas (24.75%), but in none of the benign breast specimens (p < 0.001). Out of the 25 positive cases, 14 were HPV-16 positive (56%) and 10 were HPV-18 positive (40%). An original finding was the detection of both HPV-16 and -18 in a single tumor (4%). The amplified viral sequences confirmed the presence of HPV-16 and -18. No correlation between the presence of HPV DNA and specific prognostic predictors for the disease outcome was observed. Our results suggest that the presence in the breast of either HPV-16 or -18 might be related to development of the malignant phenotype. Further studies are warranted.

MnSOD Gene Polymorphism Association with Steroid-Dependent Cancer

Oxidative stress enhances carcinogenesis due to DNA damage. Manganese superoxide dismutase (MnSOD) Val16Ala polymorphism has been recently associated with breast and prostate cancer. The role of oxidative stress in male breast cancer is poorly investigated due to the low prevalence of this neoplasia. We studied the relationship between prostate cancer (PC), male (MBC) and female breast cancer (FBC) and this polymorphism in a case–control study. Human genetic polymorphism Val16Ala of MnSOD was obtained from blood and paraffin-embedded tumor samples. The polymorphism was determined in 11 cases of MBC, 51 cases of PC, 89 cases of FBC and 372 age-adjusted healthy controls by polymerase chain reaction–restriction fragment length polymorphism techniques using restriction enzyme Hae III. Chi-square or Fisher test were used to compare the MnSOD frequency distribution. The observed genotypic frequencies of all samples were AA = 9.6% (n = 50), VV = 25.4% (n = 133) and AV = 64% (n = 340), all at Hardy–Weinberg equilibrium. Breast and prostate cancer risk was elevated in male and female patients with the Ala/Ala genotype compared to controls (p = 0.006, odds ratio = 2.5, 95% confidence interval 1.393–4.541). Even though the frequency of the Ala allele was low (9.6%) in the studied population, these data support the hypothesis that MnSOD and oxidative stress play a significant role in breast cancer risk both in males and females and also brings new information on the role of this polymorphism in prostate cancer. This is the first study which provides some evidence that genetic polymorphism in the MnSOD gene may be associated with an increased risk of male breast cancer. Studies with a larger sample size are needed to confirm the findings.

Immunohistochemical expression of p16 and p53 in vulvar intraepithelial neoplasia and squamous cell carcinoma of the vulva.

This study was undertaken to examine the expression of p16 and p53 in vulvar intraepithelial neoplasia (VIN) and squamous cell carcinoma (SCC) of the vulva. We also analyzed the relationship between p16 and p53 immunoexpression in women younger vs. older than 55 years of age. Seventyseven histologic samples of vulvar tissue, treated surgically between June 2000 and November 2004 at the Complexo Hospitalar Santa Casa (Porto Alegre, Brazil), were investigated. We analyzed 28 cases of VIN, 37 cases of SCC and 12 normal vulvar tissues. The percentage of immunohistochemical positivity for p16 had the following distribution across the groups: VIN: 21.4% (6/28), cancer: 24.3% (9/37) and control: absent (p=0.202). p53 expression showed the following percentages: VIN: 60.7% (17/28), cancer: 18.9% (7/37) and control: 8.3% (1/12) (p=0.01). p16 expression in the cancer group (mean age: 63.4 years) was positive in 6 and 3 cases of women younger or older than 55 years, respectively (54.5% vs. 11.5%, p=0.01). p53 expression was not detected in young females with cancer, while it was expressed in 7/26 (26.9%) cases of the group of females older than 55 years of age (p=0.08). Our results suggest an increase in the immunohistochemical expression of p16 protein in young women with squamous cell carcinoma of the vulva, and a possible association with a low expression of p53. (Pathology Oncology Research Vol 12, No 3, 153–157)

Polymorphism (ALA16VAL) correlates with regional lymph node status in breast cancer

We studied the possible association between Ala16Val manganese-dependent superoxide dismutase (MnSOD) gene genotypes and breast cancer lymph node status because previous investigations suggested an association between the AA genotype and breast cancer. We included 281 women (188 controls and 93 cases of invasive breast cancer with axillary lymph node metastasis (LN+) and without lymph node metastasis (LN-). DNA was extracted from paraffin-embedded tumor tissue or peripheral blood leukocytes, and MnSOD polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism techniques. In addition, the immunohistochemical profile (p53, Ki-67 and estrogen/progesterone receptors) was also compared between invasive breast cancer groups and different MnSOD genotypes. The frequency of the VV genotype was higher in the LN+ group than in the control and LN- groups (chi(2)=5.081, P=0.02). Subjects with LN+ breast cancer (LN+ group) showed a higher incidence of VV genotype carriers associated with positive Ki-67 marker. Subjects with LN+ breast cancer (LN+ group) showed a higher incidence of VV genotype carriers associated with negative p53 marker. Despite the fact that the AA genotype is well established as being associated with an increased risk of breast cancer, the VV genotype may be associated with a higher metastatic potential, suggesting that MnSOD imbalance is the condition associated with carcinogenesis.

Papillary thyroid carcinoma: does the association with Hashimoto’s thyroiditis affect the clinicopathological characteristics of the disease?

INTRODUCTION Papillary carcinoma is the most common malignant thyroid neoplasm. The effect of the concurrent presence of Hashimotos thyroiditis and papillary thyroid carcinoma remains controversial. OBJECTIVE To evaluate the association between Hashimotos thyroiditis and clinicopathological parameters in thyroid papillary carcinoma cases, based on an historical institutional cohort analysis. METHODS Cross-sectional study obtained from a historical cohort, including all cases submitted to thyroidectomy for papillary thyroid carcinoma in a single institution during an 11-year period study. RESULTS A total of 417 patients with papillary thyroid carcinoma were enrolled; 148 (35.4%) also had Hashimotos thyroiditis. A female predominance among cases associated to Hashimotos thyroiditis was observed. The thyroid tumor, in cases associated with Hashimotos thyroiditis, had a smaller mean diameter, lower frequency of extra-thyroid extension, and earlier clinicopathological staging. CONCLUSIONS A high proportion of papillary thyroid carcinoma cases are associated with Hashimotos thyroiditis. There are associations among these cases with several histopathological factors already recognized for their prognostic value, which by themselves could impact outcomes.

Are the Pure In Situ Breast Ductal Carcinomas and Those Associated With Invasive Carcinoma the Same

It has been reported that pure ductal carcinoma in situ of the breast has morphologic differences from the in situ component of the invasive ductal carcinoma, as well estrogen and progesterone receptors expression according to their cytokeratin expression. However, there is no data comparing other tumor markers without using the cytokeratin expression. The objective of this study is to compare the expression of estrogen receptor (ER) and progesterone receptor (PR), HER-2/neu, p53, and Ki67 between pure ductal in situ carcinoma (pDCIS) and the in situ component of the invasive ductal carcinoma (DCIS+IDC) of the breast, and the in situ component to the invasive component of the same tumor (DCIS+IDC). The immunohistochemistry expression of the tumor markers was performed in 45 cases of pDCIS and DCIS+IDC, yielding a total of 90 cases. Statistical analysis was carried out using Fisher exact test, having a P<0.05, and Kappa index (κ) to assess intratumoral concordance. In DCIS+IDC, the in situ and invasive components did not show a significant difference and Kappa index (κ) was high (0.7-1) for positive and negative expression. ER and PR were significantly different between the pDCIS and DCIS+IDC (ER: 86.7 vs. 66.7% P=0.04; PR: 80% vs. 55.6% P=0.02). These findings suggest that in situ component of DCIS+IDC and pDCIS are distinct conditions.

Expression of Ki-67 and squamous intraepithelial lesions are related with HPV in endocervical adenocarcinoma.

To estimate the association between human papillomavirus (HPV) status and the expression of p53, Ki-67 and bcl-2 in cases of endocervical adenocarcinoma, and the relation with squamous intraepithelial lesions (SIL) and age, 229 cases were selected, treated between 1995 and 2003 in the Hospital Nossa Senhora da Conceição. All samples were evaluated by polymerase chain reaction to determine HPV status. Immunohistochemical technique was used to investigate the expression of p53, Ki-67 and bcl-2. The joint occurrence of endocervical adenocarcinoma and SIL were estimated too. In the 229 evaluated cases, 182 cases (79.48%) were associated with the presence of the HPV. The most common types were HPV18 (93 cases-51.09%) and HPV16 (62 cases-34.06%). Expression of Ki-67 (p=0.009) and the presence of SIL (p=0.018) were associated to HPV infection. Expression of p53 (p=0.647) and bcl-2 (p=0.671) were not related to HPV status. The mean age of the patients was 53.2 years, without clear correlation between age group and HPV (p=0.095). The presence of HPV, especially type 18 in endocervical adenocarcinoma suggests that this agent can be an important cofactor in the development and progression of glandular neoplasms of the uterine cervix. The joint occurrence of endocervical adenocarcinoma and SIL may support this hypothesis. HPV may promote an increased proliferation index in endocervical adenocarcinoma, shown by the expression of Ki-67.

Association of manganese superoxide dismutase gene polymorphism (Ala-9Val) and breast cancer in males and females

INTRODUCAO: Uma das diversas rotas metabolicas envolvidas no processo de carcinogenese da mama e o polimorfismo Ala-9Val do gene da superoxido dismutase dependente de manganes, cuja associacao com o aumento do risco de câncer de mama em mulheres ja e bem estabelecida na literatura. Contudo, nao existem estudos envolvendo esse polimorfismo no carcinoma de mama em homens, principalmente devido a baixa prevalencia dessa neoplasia. O objetivo deste estudo e analisar o polimorfismo da MnSOD em uma populacao de homens e mulheres com câncer de mama no sul do Brasil, comparando tais achados com controles saudaveis. METODOS: Foi realizado um estudo de caso-controle em cem pacientes com câncer de mama (11 homens e 89 mulheres) e 370 controles saudaveis. O DNA foi extraido do tecido tumoral emblocado em parafina. O polimorfismo da MnSOD foi determinado por tecnicas de PCR-RFLP usando a enzima de restricao Hae III. O teste do qui quadrado foi usado para comparar a distribuicao das frequencias dos polimorfismos. RESULTADOS: As frequencias genotipicas dos pacientes com câncer de mama foram AA = 15%; AV = 56%; VV = 29% e dos controles AA = 6,5%; AV = 68,1% e VV = 25,4%. Os pacientes com câncer de mama, tanto as mulheres como os homens, apresentaram frequencias significativamente mais elevadas do genotipo AA quando comparadas aos controles (p = 0,035), sugerindo associacao forte desse genotipo com o câncer de mama. O intervalo de confianca foi de 1,393-4,541 (95%) e o risco encontrado foi de 2,15 para individuos portadores do genotipo AA, quando comparados com os controles que tinham os genotipos VV e AV da MnSOD. DISCUSSAO: Esses resultados confirmam a associacao ja estabelecida do genotipo da MnSOD AA com câncer de mama em mulheres e indicam distribuicao de frequencia similar e risco aumentado na populacao masculina.

Analysis of pattern of occurrence of thyroid carcinoma between 2001 and 2010

INTRODUCTION An ongoing discussion is found in medical literature about the reasons for changes in thyroid carcinoma incidence patterns over the last decades. OBJECTIVE To analyze the clinical and pathological characteristics of thyroid carcinoma cases over a decade. METHODS Cross-sectional study over an historical cohort. Medical records of 628 thyroid cancer cases in a single center were reviewed. 597 patients were included. Microcarcinoma cases were selected for a qualitative analysis phase, in which medical records were reviewed for better understanding of thyroid nodule and thyroid cancer diagnosis process. RESULTS An increase in the proportion of cases with thyroid cancer diagnosis was observed throughout the decade; new cases were predominantly tumors of less than 2 cm, with histopathological signs of low aggressiveness. There was an increase in proportion of cases with malignant cytological results among microcarcinomas. CONCLUSION There is a trend for increase in thyroidectomies due to cancer in this institution, with proportional increment of cases with histopathological characteristics indicative of early disease. Among microcarcinomas, there is an increasing group represented by cancer cases that were not incidentally diagnosed, related to an enhancement in preoperative diagnostic methods.

Relação entre escore de Gleason e fatores prognósticos no adenocarcinoma acinar de próstata

INTRODUCTION: The prostate adenocarcinoma corresponds to one of the most frequent malignant neoplasias affecting 60 to 80 year old men. Some histopathological characteristics of the tumor, such as degree of differentiation, capsular invasion and Gleason score, may be correlated with the prognosis of the disease. OBJECTIVES: To assess the association of Gleason score and prognostic factors in cases of prostatic adenocarcinoma. MATERIAL AND METHOD: The study evaluated one hundred and eighteen specimens of radical prostatectomy from the laboratory of pathology of ULBRA between 2003 and 2008. In each case, the following data were recorded: age, laterality, staging, total score, primary and secondary patterns of Gleason, degree of differentiation, presence of capsular, angiolymphatic and perineural invasion and extraprostatic extension. Chi-square test and Fischers exact test with significance < 5% were used. RESULTS: The mean age was 63.86. There was a prevalence of Gleason score 6 (55 cases - 46.61%), bilaterality (72 cases - 61.02%) and staging T2c (57 cases - 48.31%). Gleason score was associated with age (0.001), laterality (p < 0.001), capsular invasion (p < 0.005), angiolymphatic invasion (p < 0.001), perineural invasion (p < 0.05), extraprostatic invasion (p < 0.001) and staging (0.001). There was a significant association among capsular invasion, age (p < 0.01) and perineural invasion (p < 0.001). CONCLUSION: In prostate adenocarcinomas, the determination of Gleason score corresponds to one of the main histological factors to be described, showing significant relation with other anatomopathological criteria, such as extension, perineural and angiolymphatic invasion and staging.