Expression of CK7 and CDKN2 in Cervical Intraepithelial Neoplasia and Correlation with Clinical Outcome.

AIM: To evaluate cyclin-dependent kinase inhibitor 2A (CDKN2A) and cytokeratin 7 (CK7) expression in cervical intraepithelial neoplasia (CIN) formalin-fixed samples. MATERIALS AND METHODS: Staining with antibody clones G175-405 for CDKN2A and OV-TL 12/30 for CK7 were evaluated and the detection of protein expressions were compared in 147 patients with CIN. RESULTS: Clinical follow-up of patients with CIN1 and CIN2 showed that most patients had a favorable outcome. Single CDKN2A or CK7 expression and their combined expression had a greater sensitivity and negative predictive value in CIN1, corresponding to the non-development of the disease. The positive predictive value of CDKN2A was greater than that of CK7. Combined expression of CDKN2A and CK7 showed that the sensitivity, specificity, positive predictive values, and negative predictive values had their maximum index in the CIN1 group. Analysis of combined expression of CDKN2A and CK7 showed that 85.7% of patients presented unfavorable clinical outcomes, with positive expression for both markers identified in CIN2. CONCLUSION: Combined expression of CK7 and CDKN2A was associated with a better diagnosis of CIN, and negative expression in CIN1/2 groups had a greater negative predictive value for patient clinical outcome.

Dual p16 and Ki-67 Expression in Liquid-Based Cervical Cytological Samples Compared to Pap Cytology Findings, Biopsies, and HPV Testing in Cervical Cancer Screening: A Diagnostic Accuracy Study.

OBJECTIVE: Our objective was to verify the sensitivity and specificity of dual immunocytochemistry staining for p16 and Ki-67 in liquid-based samples (the "dual" assay) for cervical lesion screening, compared to biopsy findings and human papillomavirus (HPV) DNA molecular detection. STUDY DESIGN: Sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values for the "dual immunocytochemistry assay" were calculated and compared to histopathological results and to high-risk HPV DNA detection in adult women or teenagers submitted to cervical cancer screening. RESULTS: A total of 151 women were included. The majority (96.2%) of those with negative dual assay results had lower biopsy grades (p < 0.001). Women with cytology results suggestive of cervical cancer had positive dual immunocytochemistry assay results more frequently (p < 0.001), and these positive results were also significantly associated with biopsy findings (p < 0.001) and with high-risk genotype HPV infection (p = 0.007). Specificity and PPV for the dual assay were 0.972 (0.855-0.999) and 0.800 (0.284-0.995), respectively, and 1.000 (0.590-1.000) and 1.000 (0.631-1.000) for HPV detection. CONCLUSIONS: The dual immunocytochemistry assay had high specificity and PPV. It reveals a persistent HPV infection, avoiding the need for new tissue collections for biopsies or hybrid capture.

DLG1 polarity protein expression associates with the disease progress of low-grade cervical intraepithelial lesions.

Human Discs large tumour suppressor (DLG1) participates in regulating cell polarity and proliferation, suggesting an important connection between epithelial organization and cellular growth control. However, it was demonstrated that DLG1 could acquire oncogenic attributes in some specific contexts. In this work, we evaluated the expression of DLG1 and its contribution to the progress of cervical lesions in order to investigate a potential role of this polarity protein in human oncogenic processes. We analyzed cervical biopsies from women with low-grade squamous intraepithelial lesion (LSIL) diagnosis (n=30), for DLG1 expression by immunohistochemistry. These results were correlated with the clinical monitoring of the patients during a 24-month follow-up period. Our data indicate that while all LSIL patients with a DLG1 staining pattern similar to normal tissues are significantly more likely to regress (n=23, Pattern I), all LSIL biopsy specimens showing a diffuse and intense DLG1 staining likely progress to high-grade lesions (n=4, Pattern II). Finally, all persistent LSIL analyzed showed an undetermined DLG1 staining, with a diffuse distribution without a strong intensity (n=3, Pattern III). We found a significant association between the expression pattern of DLG1 and the evolution of the lesion (p<0.00001). This work contributes to the knowledge of DLG1 biological functions, suggesting that its expression may have an important role in the progression of early dysplastic cervical lesions, giving prognostic information.

Immunohistochemical Expression of VEGF and Podoplanin in Uterine Cervical Squamous Intraepithelial Lesions.

VEGF and podoplanin (PDPN) have been identified as angiogenesis and/or lymphangiogenesis regulators and might be essential to restrict tumor growth, progression, and metastasis. In the present study, we evaluate the association between the expression of these markers and CIN grade. Immunohistochemistry was performed in 234 uterine cervical samples using conventional histologic sections or TMA with the monoclonal antibodies to VEGF (C-1 clone) and podoplanin (D2-40 clone). Positive-staining rates of VEGF in 191 CIN specimens were significantly associated with histological grade (P < 0.001). Negative and/or focal immunostaining for PDPN were more frequent in CIN 3 (P = 0.016). We found that patients with CIN 3 more frequently had strong and more diffuse staining for VEGF and diminished staining for PDPN (P = 0.018). Strong and more diffuse VEGF immunoexpressions in CIN 2 and CIN 3 were detected when compared to CIN 1. Negative and/or focal PDPN immunoexpression appear to be more frequent in CIN 3. Moderate to strong VEGF expression may be a tendency among patients with high-grade lesions and diminished PDPN expression.

[Expression of TREM-1 in patients with invasive cervical cancer and precursor lesions]. / Expresión de TREM-1 en pacientes con cáncer cervical invasor y lesiones precursoras.

BACKGROUND: It has been shown that the TREM-1 glycoprotein belongs to the immunoglobulin superfamily that induces secretion of various proinflammatory cytokines. The aim of this study was to measure the expression of TREM-1 in patients with cervical cancer. METHODS: In this cross-sectional study we included four groups of patients: GI: women with low-grade squamous intraepithelial lesion (IL) (n = 15 p / g); GII: patients with high-grade squamous IL (n = 9 w / g); GIII: patients with invasive cervical cancer (n = 9 p / g) and GIV: healthy patients (n = 15 p / g). In all patients the expression of TREM-1 and the Average Fluorescence Index (AFI) in neutrophils and monocytes were measured, as well as levels of leukocytes, neutrophils and monocytes. We used Student's t test for independent samples. For these variables, we applied Mann-Whitney rank-sum, ANOVA, and Tukey tests. Chi square test was used for qualitative variables. RESULTS: The percentages of TREM-1 expression in neutrophils and monocytes, plus the AFI in neutrophils in the 4 groups was not significantly different. The AFI of TREM-1 in monocytes was significantly different when comparing group II and group III versus group IV (p < 0.02). There was also no significant difference when comparing the mean values of leukocytes, neutrophils and monocytes in the different groups. CONCLUSION: This study shows increased expression of TREM-1 in monocytes from patients with advanced cancer.

Introducción: se ha demostrado que la glicoproteína TREM-1 pertenece a la superfamilia de las inmunoglobulinas que induce la secreción de varias citocinas proinflamatorias. El objetivo de este trabajo fue medir la expresión de TREM-1 en pacientes con cáncer cervical. Métodos: en este estudio transversal analítico incluimos 4 grupos de pacientes: GI: mujeres con lesión intraepitelial (LI) escamosa de bajo grado (n = 15 p/g); GII: pacientes con LI escamosa de alto grado (n = 9 p/g); GIII: pacientes con cáncer cervical invasor (n = 9 p/g), y GIV: pacientes sanas (n = 15 p/g). En todas las pacientes se midió la expresión de TREM-1 y el Índice Medio de Fluorescencia (IMF) en neutrófilos y monocitos, así como los niveles de leucocitos, neutrófilos y monocitos. Usamos t de Student para muestras independientes. Para estas mismas variables, aplicamos prueba de suma de rangos de Mann-Whitney, ANOVA y Turkey. Para las variables cualitativas se utilizó la prueba de Chi cuadrada. Resultados: los porcentaje de expresión de TREM-1 en neutrófilos y monocitos, además del IMF en neutrófilos en los 4 grupos, no fue significativamente diferente. El IMF de TREM-1 en monocitos fue significativamente diferente al comparar el grupo II y grupo III frente al grupo IV (p < 0.02). Tampoco hubo diferencia significativa al comparar los valores promedio de leucocitos, neutrófilos y monocitos en los diferentes grupos. Conclusión: este estudio documenta una mayor expresión de TREM-1 en monocitos de pacientes con cáncer avanzado.

Progressive loss of E-cadherin immunoexpression during cervical carcinogenesis.

PURPOSE: To investigate E-cadherin immunoexpression during cervical carcinogenesis. METHODS: We assessed the immunohistochemical expression of E-cadherin in squamous intraepithelial lesions (SIL - 52 cases), squamous cell carcinoma (SCC) of the uterine cervix (23 cases) and also in eight cases of cervicitis. RESULTS: The results show very different E-cadherin membrane expression levels when cervicitis (88%), SILs (73%) and SCC (17%) were compared. In SILs, higher E-cadherin loss was seen in less differentiated cells in the basal third of the epithelium. This study suggests that the absence of E-cadherin expression in the membrane is a molecular event that is observed more often in SCC of the uterine cervix than in SILs or cervicitis. CONCLUSIONS: E-cadherin is an essential molecule during the process of cervical carcinogenesis and in this context exhibits a different expression pattern according to the epithelial thickness layer.

Progressive loss of E-cadherin immunoexpression during cervical carcinogenesis

PURPOSE: To investigate E-cadherin immunoexpression during cervical carcinogenesis. METHODS: We assessed the immunohistochemical expression of E-cadherin in squamous intraepithelial lesions (SIL - 52 cases), squamous cell carcinoma (SCC) of the uterine cervix (23 cases) and also in eight cases of cervicitis. RESULTS: The results show very different E-cadherin membrane expression levels when cervicitis (88%), SILs (73%) and SCC (17%) were compared. In SILs, higher E-cadherin loss was seen in less differentiated cells in the basal third of the epithelium. This study suggests that the absence of E-cadherin expression in the membrane is a molecular event that is observed more often in SCC of the uterine cervix than in SILs or cervicitis. CONCLUSIONS: E-cadherin is an essential molecule during the process of cervical carcinogenesis and in this context exhibits a different expression pattern according to the epithelial thickness layer. .

MMP-9 expression increases according to the grade of squamous intraepithelial lesion in cervical smears.

Studies about cervical carcinogenesis have demonstrated the increased expression of matrix-metalloproteinase (MMP) according to the grade of cervical intraepithelial lesions. Considering the importance of innovative techniques to introduce noninvasive and rapid diagnoses for patients, this study aimed to perform MMP-9 immunocytochemistry in cervical smears according to the cytopathological diagnoses, in order to monitor MMP activity in cervical smears. This cross-sectional study investigated the expression of MMP-9 in normal cervical smears, inflammatory cervical smears, squamous intraepithelial lesions, and cervical carcinoma. Cervical smears from 630 women were collected for cytopathological diagnoses and immunocytochemistry. Women with squamous intraepithelial lesions showed an increase in MMP-9 expression, with moderate to intense staining occurring with increasing cervical lesion grade. The prevalence of moderate to intense MMP-9 staining was 9% in normal cervical smears, 12% in cervical inflammation, 24% in low-grade squamous intraepithelial lesion (LSIL), 92% in high-grade squamous intraepithelial lesions (HSIL) and 100% in cervical carcinoma cases. In the specific case of LSIL, we found that association with MMP-9 is more evident when there is the simultaneous presence of an infectious agent. Thus, the expression of MMP-9 in cervical smears increases according to the grade of cervical lesion and LSIL in the presence of infectious agents showed higher MMP-9 expression than women with LSIL without infectious agents.