Persistent High-Risk HPV Infection and Molecular Changes Related to the Development of Cervical Cancer.

This article is a preliminary investigational study that is aimed at giving hints about the interesting biomarkers involved in the transition process from low-grade cervix lesion to invasive cervical cancer. Our study focuses on the risk factors and tumour molecular changes in one patient. First in 1986, she was diagnosed a preinvasive cervix lesion. Then, 16 years later, she was diagnosed an invasive cervical cancer. The 2002 diagnosis was a squamous cell carcinoma of the cervix, stage IIIB (FIGO), whereas in 1986, she had been diagnosed a high-grade squamous intraepithelial cervical lesion. Retrospectively, the analysis of samples of preneoplastic lesions and invasive cervical cancer confirmed the histopathological diagnoses and detected the presence of HPV type and HPV-16 variants, as well as the overexpression of proteins such as hTERT, IGF1Rα, IGF1Rß, CAIX, and GLUT1. Finally, the Arg72Pro polymorphism was detected in TP53. The role of high-risk HPV and HPV-16 variants and of hTERT, IGF1Rα, IGF1Rß, CAIX, and GLUT1 variations seemed confirmed in the development and progression of cervical cancer. As a result, analyzing the molecular changes in one and same tumour that progresses from a low-grade cervix lesion to invasive cervical cancer could provide valuable information in order to improve detection, diagnosis, and treatment in the future.

hTERT Protein Expression in Cytoplasm and Nucleus and its Association With HPV Infection in Patients With Cervical Cancer.

BACKGROUND: Few studies have analyzed the association between human telomerase reverse transcriptase (hTERT) protein expression (nuclear and cytoplasmic localization), hTERT methylation status, and human papillomavirus (HPV) genotype infection in cervical cancer. PATIENTS AND METHODS: One hundred seventy-three patients with cervical cancer were analyzed. hTERT protein expression was detected by immunohistochemistry. hTERT DNA methylation analysis was performed using a PCR-RLB-hTERT assay, targeting two regions of the hTERT promoter. Type specific HPV infection was detected by using GP5+/GP6+PCR-RLB. RESULTS: hTERT protein expression was found in both cytoplasm and nucleus (78.0% of the samples showed a cytoplasmic localization and 79.8% had a nuclear localization). A statistically significant association was found between alpha 9 and 7 HPV species with a non-methylation pattern of the hTERT promoter and between these species and high expression of hTERT protein with nuclear localization. CONCLUSION: hTERT protein is found in both the nucleus and cytoplasm of patients with cervical cancer and confirm the relationship between the non-methylated status of hTERT promoter and some HPV species as well as the relationship between these species and hTERT protein expression.

[Immunocytochemical and molecular studies with primary cultures of molar tissue]. / Estudio inmunocitoquímico y molecular de cultivo primario de tejido molar.

INTRODUCTION: Gestational trophoblastic disease includes a group of pathologies characterized by abnormal trophoblast growth and invasion. The molecular bases of the disease are largely unknown, due in part to the lack of appropriate biological models. The insulin-like growth factor (IGF) system plays a fundamental role in the growth and development of many tissues and is involved in the progression of several diseases. OBJECTIVES: Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods. MATERIALS AND METHODS: The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression. RESULTS: Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions. CONCLUSION: By combining three methodologies-morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.

Novel predictive biomarkers for cervical cancer prognosis.

High hypoxic, glycolytic and acidosis metabolisms characterize cervical cancer tumors and have been described to be involved in chemoradioresistance mechanisms. Based on these observations, the present study assessed four selected novel biomarkers on the prognosis of locally advanced cervical carcinoma. A total of 66 patients with stage IIB/IIIB cervical cancer were retrospectively included. The protein expression levels of glucose transporter 1 (GLUT1), carbonic anhydrase 9 (CAIX) and hexokinase 1 (HKII) were investigated by immunohistochemistry on tumor biopsies, hemoglobin was measured and the disease outcome was monitored. A total of 53 patients (80.3%) presented a complete response. For these patients, the protein expression levels of GLUT1, CAIX and HKII were overexpressed. A significant difference was observed (P=0.0127) for hemoglobin levels (≤11 g/dl) in responsive compared with non-responsive patients. The expression of GLUT1 is associated with a lower rate of both overall and disease-free survival, with a trend of decreased risk of 1.1x and 1.5x, respectively. Co-expression of GLUT1 and HKII is associated with a decreased trend risk of 1.6x for overall survival. Patients with hemoglobin levels ≤11 g/dl had a 4.3-fold risk (P=0.02) in decreasing both to the rate of overall and disease-free survival. The presence of anemic hypoxia (hemoglobin ≤11 g/dl) and the expression of GLUT1 and/or HKII influence treatment response and are associated with a lower overall and disease-free survival. The present results demonstrated that these biomarkers may be used as predictive markers and suggested that these metabolic pathways can be used as potential novel therapeutic targets.

Detección del antígeno Tn en tumores epiteliales con la lectina de Vicia villosa isolectina B4 / Using Vicia villosa lectin (B4 isolectin) for detecting Tn antigen in epithelial tumours

Antecedentes. Los epítopes T, Tn y sTn, se expresan en un alto porcentaje de tumores epiteliales y pueden detectarse con anticuerpos monoclonales y lectinas. Objetivo. Evaluar diferencias de expresión del antígeno Tn en cortes histológicos de epitelios no neoplásicos y tumores epiteliales mediante isolectina B4 de Vicia villosa. Material y métodos. Se evaluaron semicuantitativamente localización, intensidad y porcentaje de expresión del antígeno en carcinomas in-situ e infiltrantes y epitelios no neoplásicos de cérvix, seno y urotelio, mediante isolectina B4. Resultados La expresión de Tn en cérvix predominó en membrana de células no neoplásicas y citoplasma de células tumorales; su intensidad fue mayor en carcinomas in-situ e infiltrantes comparado con epitelio no neoplásico aunque en este el porcentaje de expresión fue mayor. En seno, la expresión de Tn fue predominantemente citoplasmática con intensidad similar, el porcentaje de expresión fué mayor en carcinomas ductales in-situ e infiltrantes. En urotelio no neoplásico y tumoral la expresión de Tn predominó en citoplasma; la intensidad y el porcentaje de expresión fueron mayores en neoplasias no invasivas de bajo y alto grado, mientras que en urotelio no neoplásico fue baja y no hubo tendencia definida en tumores infiltrantes. Conclusiones. La detección del antígeno Tn mediante la lectina VVB4 mostró una mayor extensión de marcación en carcinomas ductales de seno en relación con el epitelio no neoplásico, pero no mostró una tendencia definida entre el tejido normal, ni diferentes etapas del desarrollo de los tumores de cérvix y urotelio. Estos hallazgos pueden atribuirse a la heterogeneidad de los procesos carcinogénicos o a que la especificidad de la lectina VVB4 no está restringida a este antígeno.

Estudio inmunocitoquímico y molecular de cultivo primario de tejido molar / Immunocytochemical and molecular studies with primary cultures of molar tissue

Introducción. La enfermedad trofoblástica gestacional comprende un conjunto de patologías caracterizadas por crecimiento e invasión anómalos del trofoblasto. Las bases moleculares de esta patología son desconocidas, en parte por la dificultad para disponer de modelos biológicos adecuados. Se plantea que el sistema de factores de crecimiento similares a la insulina puede tener un papel fundamental en el desarrollo de la enfermedad. Objetivo. Caracterizar cultivos primarios de placentas de primer trimestre provenientes de pacientes con mola hidatidiforme completa y aborto espontáneo no molar mediante morfología, inmunocitoquímica y expresión diferencial de algunos genes del sistema de factores de crecimiento similares a la insulina. Materiales y métodos. Se empleó inmunocitoquímica para determinar células trofoblásticas y detección por transcripción reversa y reacción en cadena de la polimerasa de genes del sistema de factores de crecimiento similares a la insulina asociados al tipo celular. Resultados. La morfología evidenció heterogeneidad de los cultivos, incluidas células mesenquimales, trofoblásticas y de decidua. El contenido de células de trofoblasto con citoqueratina-7 (marcador específico) estuvo entre 16 y 37 por ciento. La expresión de genes corroboró la presencia de trofoblasto por medio del ARNm del factor II de crecimiento similar a la insulina, en tanto que los transcritos de la hormona de crecimiento variante evidenciaron la presencia de sincitiotrofoblasto. El factor I de crecimiento similar a la insulina y la proteína de unión tipo 1 se relacionaron con células mesenquimales y de decidua. Se observó una mayor expresión del factor II de crecimiento similar a la insulina en tejidos molares en comparación con aborto no molar. Conclusiones. Los resultados mostraron la utilidad de combinar tres metodologías, morfología, inmunocitoquímica y expresión de genes, como herramientas para la caracterización y seguimiento de cultivos placentarios a partir....

Introduction. Gestational trophoblastic disease includes a group of pathologies characterized by abnormal trophoblast growth and invasion. The molecular bases of the disease are largely unknown, due in part to the lack of appropriate biological models. The insulin-like growth factor (IGF) system plays a fundamental role in the growth and development of many tissues and is involved in the progression of several diseases. Objectives. Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods. Materials and Methods. The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression. Results. Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions. Conclusion. By combining three methodologies—morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.