HIF-1α expression and high microvessel density are characteristic features in serrated colorectal cancer.

Serrated colorectal adenocarcinoma (SAC) is a morphologically distinct subtype of colorectal cancer (CRC), in which increased HIF-1α mRNA expression and HIF-1α protein stabilization are typical features. Here we aimed to further elucidate HIF-1α protein expression in serrated and non-serrated colorectal carcinomas (CRCs) and their precursor lesions and its association with vascular endothelial growth factor (VEGF) and microvascular density (MVD). HIF-1α and VEGF expressions were determined immunohistochemically in 134 serrated polyps (SPs), 104 non-serrated adenomas (NSAs), 81 SACs, and 74 matched conventional adenocarcinomas (CCs) and were correlated with morphology, clinicopathological features, and MVD. In premalignant lesions, both HIF-1α and VEGF were expressed in the vast majority of SPs and NSAs. In CRCs, HIF-1α protein was also present in 77.8 % of SACs, while only 20.3 % of CCs were HIF-1α proficient. MVD was significantly higher in SACs, but the serrated morphology was the only significant predictor of MVD in CRC in multivariate analyses. HIF-1α protein is often stabilized in well-vascularized SACs, suggesting hypoxia-independent stabilization of HIF-1α. Moreover, HIF-1α stabilization did not associate with oncogenic activation of BRAF or KRAS or Von Hippel-Lindau (VHL) mutation. Prevalent HIF-1α expression in SAC and its precursors support the importance of HIF-1α-mediated pathways for the serrated route of colorectal carcinogenesis.

Tumor rabdoide extrarrenal maligno de colon: presentación de 3 casos y revisión de la literatura / Malignant extrarenal rhabdoid tumour of the colon: a report of 3 cases and a review of the literature

El tumor rabdoide extrarrenal maligno (TREM) de colon es una neoplasia maligna poco frecuente de la que solo hay 12 casos publicados en la bibliografía. Los casos suelen presentar unas características clínicas (pacientes de edad avanzada, tumores de gran tamaño y comportamiento biológico agresivo), histológicas (características típicas de tumor rabdoide) e inmunohistoquímicas (expresión de vimentina y cóctel de citoqueratinas) comunes. Presentamos 3 casos de TREM colónicos con características acordes a las observadas en la bibliografía. Uno de los 3 casos es una probable metástasis de un carcinoma de células renales con características rabdoides. A estos casos se les ha realizado un amplio estudio inmunohistoquímico, así como la determinación del estado de mutaciones en KRAS y BRAF para comparar estos hallazgos con los descritos en la bibliografía y realizar un diagnóstico diferencial con posibles neoplasias colónicas primarias o metastásicas que pueden presentar un aspecto semejante (AU)

Malignant extrarenal rhabdoid tumour (MERT) of the colon is rare, with only 12 cases published to date. Clinically, they usually occur in elderly patients and present as large, aggressive tumours; their histopathology is that typical of rhabdoid tumours and their immunohistochemistry shows expression of vimentin and cytokeratines. We present 3 cases of colonic MERTs with features similar to previously published cases. One was possibly a metastasis of a renal cell carcinoma with rhabdoid features. We carried out a extensive immunohistological analysis of all 3 cases as well as testing for KRAS and BRAF mutations. Thus we could not only compare our findings with those of the other known cases but also make a differential diagnosis with primary colonic neoplasmas and metastases of similar appearance (AU)

Biomarkers for the identification of precursor polyps of colorectal serrated adenocarcinomas.

BACKGROUND: In contrast to conventional colorectal carcinomas (CCs), which develop through a so-called chromosome instability or suppressor phenotype pathway, the sequence of events leading from precursor polyps/adenomas to serrated adenocarcinomas (SACs), which are more aggressive and exhibit a poorer survival than CCs, is as yet not clearly defined. Here, we aimed at detecting protein and DNA biomarkers for SAC in a series of primary colorectal polyps. METHODS: In total 303 colorectal polyps were included: 121 serrated polyps (33 hyperplastic polyps, 37 sessile serrated adenomas (SSA), 51 traditional serrated adenomas (TSA)), 143 conventional polyps (72 tubular polyps, 34 tubulovillous polyps, 37 villious adenomas), and 39 bi-phenotypic serrated-conventional polyps. The protein biomarkers tested were deduced from previously published SAC and CC expression profiling studies. A representative subset of 106 polyps was selected for DNA biomarker analyses, i.e., proto-oncogene mutation and microsatellite instability (MSI) status. In order to confer proper weight to each biomarker, a multivariate logistic regression model was employed. RESULTS: We found that serrated and conventional polyps differed in most of the SAC biomarkers tested. Of these biomarkers, FSCN1 showed the largest difference in expression (p = 0.0001). Despite sharing a serrated morphology, we found that SSAs and TSAs differed considerably with respect to anatomical location, expression of EPHB2 and PTCH1, presence of the V600E BRAF mutation and MSI status. Logistic regression analysis revealed that SSA was the polyp type that shared most biomarkers with SAC. CONCLUSION: Based on the shared presence of protein and molecular biomarkers, especially FSCN1 expression, SSA may serve as a precursor lesion of SAC. Biomarker assessment may help in discerning colorectal carcinogenic routes with distinct prognostic implications.

Methylome profiling reveals functions and genes which are differentially methylated in serrated compared to conventional colorectal carcinoma.

BACKGROUND: Serrated adenocarcinoma (SAC) is a recently recognized colorectal cancer (CRC) subtype accounting for 7.5-8.7 % of CRCs. It has been shown that SAC has a worse prognosis and different histological and molecular features compared to conventional carcinoma (CC) but, to date, there is no study analysing its methylome profile. RESULTS: The methylation status of 450,000 CpG sites using the Infinium Human Methylation 450 BeadChip array was investigated in 103 colorectal specimens, including 39 SACs and 34 matched CCs, from Spanish and Finnish patients. Microarray data showed a higher representation of morphogenesis-, neurogenesis-, cytoskeleton- and vesicle transport-related functions and also significant differential methylation of 15 genes, including the iodothyronine deiodinase DIO3 and the forkhead family transcription factor FOXD2 genes which were validated at the CpG, mRNA and protein level using pyrosequencing, methylation-specific PCR, quantitative polymerase chain reaction (qPCR) and immunohistochemistry. A quantification study of the methylation status of CpG sequences in FOXD2 demonstrated a novel region controlling gene expression. Moreover, differences in these markers were also evident when comparing SAC with CRC showing molecular and histological features of high-level microsatellite instability. CONCLUSIONS: This methylome study demonstrates distinct epigenetic regulation patterns in SAC which are consistent to previous expression profile studies and that DIO3 and FOXD2 might be molecular targets for a specific histology-oriented treatment of CRC.

Association of polymorphisms in TRAIL1 and TRAILR1 genes with susceptibility to lymphomas.

The TRAILR1/TRAIL system is implicated in the induction of the extrinsic apoptotic pathway and constitutes an emerging target in cancer therapeutics. The objective of this study is to assess lymphoma risk associated with certain polymorphisms in TRAILR1 and TRAIL1 genes. DNA was extracted from 381 subjects (190 lymphoma cases and 191 matched controls) and genotyped for polymorphisms rs20576, rs2230229 and rs20575 in TRAILR1 and rs12488654 in TRAIL gene. In contrast to TRAILR1 polymorphisms, the genotype distribution of rs12488654 in TRAIL gene was different between cases and controls, A allele carriers (CA/AA) being much more common in the cases with different lymphoma types (follicular, 45 %; diffuse large B cell, 45.2 % and Hodgkin lymphomas, 40 %) than in controls (15.7 %) (odds ratio (OR), 3.5; CI, 2.1­5.9; p<0.001; OR, 3.5; CI, 1.6­7.9; p=0.001; OR, 2.9; CI, 1.1­7.5; p=0.027, respectively). This effect was consistently independent of the association with the TRAILR1 polymorphisms studied, as demonstrated by linkage disequilibrium and haplotype analyses. This study is the first one to report an association between a TRAIL polymorphism and lymphoma risk and suggests a possible role of TRAIL in B cell lymphomagenesis.

Carcinoma colorrectal serrado: caracterización morfológica, inmunohistoquímica y molecular / Morphological, immunohistochemical and molecular features of serrated colorectal carcinoma

El carcinoma serrado (CS) es un tipo de carcinoma colorrectal (CCR) recientemente reconocido por la OMS que representa entre el 7,5 y el 9,2% de los CCR. Los criterios histológicos para su diagnóstico fueron propuestos por investigadores finlandeses y han sido validados recientemente por autores españoles. Distintos estudios demuestran que el CS tiene peor pronóstico que el CCR convencional. Además, se han estudiado factores histológicos con valor pronóstico, la expresión inmunohistoquímica de proteínas relacionadas con la gemación tumoral y reparación del ADN y la incidencia de inestabilidad de microsatélites (MSI) y de mutaciones en KRAS y BRAF. En consonancia con las observaciones clínicas, demostramos que los CS presentan características histológicas de mal pronóstico, un patrón característico de expresión de proteínas en el frente invasivo, y estatus MSI y perfil de mutaciones diferente a los CCR no serrados. Estos hallazgos sugieren que el diagnóstico correcto del CS puede tener importantes implicaciones clínicas(AU)

Serrated carcinoma (SC) is a subtype of colorectal carcinoma (CRC). Recently, the WHO estimated that it accounts for 7.5-9.2% of all CRCs. The diagnostic criteria proposed by Finnish authors, have been confirmed by Spanish research. Various studies show that SC has a worse prognosis than conventional CRC carcinoma ( CC ). Histological features with prognostic significance have been identified in SC, including the immunohistochemical expression of proteins implicated in tumour-budding formation and DNA repair, the microsatellite instability (MSI) status and the occurrence of KRAS and BRAF mutation. In agreement with clinical evidence, we have shown histologically that SC, with a distinct pattern of proteins at the invasive front and oncogene mutations, has a worse prognosis than CC. Thus, an accurate diagnosis of SC is of clinical importance(AU)

Gene amplification and immunohistochemical expression of ERBB2 and EGFR in cervical carcinogenesis. Correlation with cell-cycle markers and HPV presence.

Although the members of the epidermal growth factor receptor family ERBB2 and EGFR are important therapeutic targets in the treatment of malignant neoplasias, little is known about their role in cervical carcinogenesis. Our objective was to evaluate the dysfunction of ERBB2 and EGFR at the gene copy number and protein expression level in neoplastic lesions of the uterine cervix with the aim of obtaining information about its role in cervical carcinogenesis and their possible use as therapeutic targets in these diseases. We studied gene amplification and protein expression of ERBB2 and EGFR and their relationship with Ki67, p16 and p53 and HPV presence in 22 normal/benign (N/B) cervices, 20 low-grade squamous intraepithelial lesions (LSILs), 70 high-grade SILs (HSILs) and 32 invasive squamous cervical carcinomas (ISCCs). No cases showed selective amplification of ERBB2 or EGFR but corresponding chromosome-specific probes displayed chromosome 17 and 7 polyploidy associated with the grade of the lesion (p<0.0001 and p=0.004, respectively) and with the positive expression of Ki67 and p16 (p<0.01). Concurrent polyploidy for both chromosomes was statistically related (p<0.0001). ERBB2 immunohistochemical expression was not observed in any of the study cases except for one ISCC but EGFR was associated with higher-grade lesions (N/B plus LSIL 21.4% vs. HSIL plus ISCC 45.5%; p=0.007). No association was observed between EGFR expression and that of cell-cycle markers or HPV presence. Increased copy number of EGFR and ERBB2 is due to polyploidy of 7 and 17 chromosomes, this being a phenomenon associated with lesion severity and with an increase in the expression of cell-cycle markers. EGFR, but not ERBB2, is expressed in precursor lesions of squamous cervical neoplasia and is related to the neoplastic progression but not to proliferation marker expression and therefore ERBB2 and this calls into question the usefulness of ERBB2 as a therapeutic target.

Polymorphisms in xenobiotic metabolizing genes (EPHX1, NQO1 and PON1) in lymphoma susceptibility: a case control study.

BACKGROUND: The interplay between genetic susceptibility and carcinogenic exposure is important in the development of haematopoietic malignancies. EPHX1, NQO1 and PON1 are three genes encoding proteins directly involved in the detoxification of potential carcinogens. METHODS: We have studied the prevalence of three functional polymorphisms affecting these genes rs1051740 EPHX1, rs1800566 NQO1 and rs662 PON1 in 215 patients with lymphoma and 214 healthy controls. RESULTS: Genotype frequencies for EPHX and NQO1 polymorphisms did not show any correlation with disease. In contrast, the GG genotype in the PON1 polymorphism was found to be strongly associated with the disease (15.3% vs. 4.7%; OR = 3.7 CI (95%): 1.8-7.7; p < 0.001). According to the pathological diagnosis this association was related to follicular (p = 0.004) and diffuse large B-cell (p = 0.016) lymphomas. CONCLUSIONS: Despite the fact that further confirmation is needed, this study shows that the PON1 GG genotype in rs662 polymorphism could be a risk factor for B-cell lymphomas.

Expression profiling shows differential molecular pathways and provides potential new diagnostic biomarkers for colorectal serrated adenocarcinoma.

Serrated adenocarcinoma (SAC) is a recently recognized colorectal cancer (CRC) subtype accounting for 7.5 to 8.7% of CRCs. It has been shown that SAC has a poorer prognosis and has different molecular and immunohistochemical features compared with conventional carcinoma (CC) but, to date, only one previous study has analyzed its mRNA expression profile by microarray. Using a different microarray platform, we have studied the molecular signature of 11 SACs and compared it with that of 15 matched CC with the aim of discerning the functions which characterize SAC biology and validating, at the mRNA and protein level, the most differentially expressed genes which were also tested using a validation set of 70 SACs and 70 CCs to assess their diagnostic and prognostic values. Microarray data showed a higher representation of morphogenesis-, hypoxia-, cytoskeleton- and vesicle transport-related functions and also an overexpression of fascin1 (actin-bundling protein associated with invasion) and the antiapoptotic gene hippocalcin in SAC all of which were validated both by quantitative real-time PCR (qPCR) and immunohistochemistry. Fascin1 expression was statistically associated with KRAS mutation with 88.6% sensitivity and 85.7% specificity for SAC diagnosis and the positivity of fascin1 or hippocalcin was highly suggestive of SAC diagnosis (sensitivity = 100%). Evaluation of these markers in CRCs showing histological and molecular characteristics of high-level microsatellite instability (MSI-H) also helped to distinguish SACs from MSI-H CRCs. Molecular profiling demonstrates that SAC shows activation of distinct signaling pathways and that immunohistochemical fascin1 and hippocalcin expression can be reliably used for its differentiation from other CRC subtypes.

Microsatellite pathologic score does not efficiently identify high microsatellite instability in colorectal serrated adenocarcinoma.

The microsatellite pathologic score has been proposed as a valuable tool to estimate the probability of a colorectal cancer having high microsatellite instability; however, this score has not been tested in serrated adenocarcinoma. Our aim was to evaluate microsatellite pathologic score in serrated adenocarcinoma, conventional carcinoma, and colorectal cancer with high microsatellite instability histologic features. Eighty-nine serrated adenocarcinoma and 81 matched conventional carcinomas were tested with microsatellite pathologic score, and the results were compared with those of 24 high microsatellite instability histologic features. Validation was performed by microsatellite instability analysis. Although all colorectal cancers with high microsatellite instability histologic features rendered a more than 5.5 score, the microsatellite pathologic score performance was of lower rank in high microsatellite instability serrated adenocarcinoma because none of the cases scored above 5.5 (>77% probability of being high microsatellite instability). High microsatellite instability serrated adenocarcinoma shows pathologic features different from those observed in high microsatellite instability histologic features such as adverse prognostic histologic features at the invasive front. We describe a serrated adenocarcinoma subtype showing high microsatellite instability and some, but not all, high microsatellite instability histologic features that would not be detected if the microsatellite pathologic score cutoff is set at the highest rank. To increase microsatellite pathologic score sensitivity in serrated adenocarcinoma, we propose to set up a 2.1 cutoff score when faced by a right-sided colorectal cancer with serrated features.

Analysis of performance characteristics of five cell cycle-related immunohistochemical markers and human papillomavirus genotyping in the diagnosis of cervical squamous cell carcinoma precursor lesions.

OBJECTIVE: To study the performance values of a set of five immunohistochemical markers involved in cell cycle regulation as a potential aid in the differential diagnosis between squamous intraepithelial lesions (SILs) and normal or benign conditions of the uterine cervix. STUDY DESIGN: Results from immunohistochemical evaluation of p16, cyclin D1, p53, Ki67, and ProEx C markers and human papillomavirus genotyping were collected from a previous study performed on 37 normal or benign cervices, 39 low grade SILs and 73 high grade SILs. A multivariate analysis was used to examine the specific diagnostic value of each marker and to ascertain those most relevant for SIL diagnosis. For markers with good data fit, sensitivity, specificity, accuracy, area under the receiver operating characteristic curve, and integrated discrimination improvement, were calculated. RESULTS: Among individual markers, ProEx C showed the best specificity; p16 displayed the highest sensitivity and area under receiver operating characteristic curve for SIL diagnosis. Integrated discrimination improvement demonstrated that the p16 plus ProEx C model has better discrimination capacity than p16 plus Ki67 or ProEx C plus Ki67. CONCLUSION: Use of ProEx C alone or in combination with p16 could provide useful information for SIL diagnosis.

Immunohistochemical expression profile of ß-catenin, E-cadherin, P-cadherin, laminin-5γ2 chain, and SMAD4 in colorectal serrated adenocarcinoma.

The immunohistochemical expression of cell adhesion molecules in colorectal serrated adenocarcinoma is still unknown. The immunostaining patterns of ß-catenin, E-cadherin, P-cadherin, laminin 5γ2, and SMAD4 and their relationship to survival were studied in different tumor areas, namely, tumor center and invasive front, the latter comprising tumor bud and non-tumor bud clusters, as described in a previous study of 66 serrated adenocarcinomas and matched conventional carcinomas. Compared with conventional carcinomas, serrated adenocarcinomas showed significantly reduced nuclear ß-catenin, membranous E-cadherin, and nuclear SMAD4 but an increased cytoplasmic expression of laminin-5γ2 at the invasive front that was particularly pronounced in the tumor buds. E-cadherin loss at the invasive front was identified as an independent prognostic factor for a poorer outcome in serrated adenocarcinoma. Serrated adenocarcinoma shows a distinct immunohistochemical profile at the invasive front compared with conventional carcinoma, which may account for its less favorable outcome. The lower frequency of nuclear ß-catenin in SAC, especially in right-sided tumors, suggests that molecular mechanisms other than the canonical Wnt/ß-catenin pathway may have a role in tumor bud formation.

Role of GSTT1 and M1 null genotypes as risk factors for B-cell lymphoma: influence of geographical factors and occupational exposure.

The interrelationship between genetic susceptibility and carcinogenic exposure is important in the development of haematopoietic malignancies. Both factors need to be considered to enable assessment of disease risk associated with a given individual under certain environmental conditions. GSTT1 and GSTM1 are two genes whose proteins are involved in the detoxification of potential carcinogens. We have studied the prevalence of GSTT1 and GSTM1 null polymorphisms using a novel PCR multiplex protocol in a group of 158 patients with B-cell lymphoma (BCL, 138 with non-Hodgkin lymphoma and 20 with Hodgkin lymphoma) and 214 healthy controls. A questionnaire regarding occupational exposure and lifestyle factors was also completed by both groups. GSTM1 null genotype showed no significant differences between patients and controls (46.9% and 55.6%, respectively). In contrast, GSTT1 null genotype was observed in 25.3% of patients and 15.4% of controls (P=0.013; OR=1.85; CI (95%):1.11-3.09), suggesting a role for the GSTT1 null genotype in the development of BCL. This effect was even more evident in females (27.5% vs. 14%: P=0.014). No significant association was observed between GST genotypes and disease risk in relation to smoking or occupational exposure.

Tumour budding and other prognostic pathological features at invasive margins in serrated colorectal adenocarcinoma: a comparative study with conventional carcinoma.

AIMS: To assess the incidence of tumour budding (TB), cytoplasmic pseudo-fragments (CyPs), tumour growth pattern (TGP) and peritumoural lymphocytic infiltration (PLI) in a series of serrated adenocarcinoma (SAC) and conventional carcinomas (CCs) of the colorectum in order to ascertain whether such features could explain the worse prognosis of SAC and whether they have prognostic value in SACs. METHODS AND RESULTS: Tumour budding, CyPs, TGP and PLI were evaluated in 81 SACs and 81 matched CCs. Kaplan-Meier survival curves and Cox logistic regression analysis were obtained for histological parameters. SACs had more high-grade (HG) TB (HG-TB) (69.1%), HG-CyPs (47%), infiltrative TGP (42%) and weak PLI (W-PLI) (65.4%) than CCs (40.7%, P = 0.0003; 19.7%, P = 0.0002; 29.7%, P = 0.07; 45.7%, P = 0.0087). SACs with HG-TB (P = 0.017), HG-CyPs (P = 0.045), infiltrating TGP (P < 0.001) and W-PLI (P = 0.04) had a worse 5-year survival, as had SACs with infiltrating TGP (P = 0.047) and W-PLI (P = 0.04) compared with CCs. For SACs, infiltrative TGP and W-PLI were independent prognostic parameters on multivariate analysis, as was location and regional node status. CONCLUSION: Compared to CC, SAC displayed more HG-TB, HG-CyPs and fewer PLI at the invasive margins and this may account for its poorer clinical outcome. TB, CyPs, TGP and PLI are useful histological prognostic aids in SAC.

Comparison of allelic discrimination by dHPLC, HRM, and TaqMan in the detection of BRAF mutation V600E.

The V600E mutation in the BRAF oncogene is associated with colorectal carcinomas, with mismatch-repair deficiency and, recently, with nonresponse to epidermal growth factor receptor inhibitor therapy. The use of reliable techniques for its detection is important. The aim of our study was to compare the performance characteristics in V600E detection of denaturing high-performance liquid chromatography (dHPLC) and high-resolution melting (HRM) with TaqMan allelic discrimination as well as direct-sequencing methods in a series of 195 colorectal paraffin-embedded specimens up to the age of 15 years. The effectiveness for obtaining results on mutation status was best using TaqMan (96.9%), followed by dHPLC (93.3%), HRM (88.7%), and sequencing (88.2%). In general, TaqMan was best for analyzing older tissues, whereas sequencing was the least efficient. Heterozygotic V600E was detected in 11.6%, 9.9%, 11.6%, and 9.9% of tissues using TaqMan, dHPLC, HRM, and sequencing, respectively. Result concordances between dHPLC and TaqMan or sequencing were excellent (κ = 0.9411 and κ = 0.8988, respectively); for HRM, the concordances were good (κ = 0.7973 and κ = 0.7488, respectively). By using DNA dilutions from tumor tissue, a minimum of 10% of V600E harboring cancer content was required for the analysis by dHPLC and HRM. dHPLC could detect four non-V600E mutations, whereas HRM detected one. Our results indicate that dHPLC and HRM are techniques that can be reliably used for the detection of the BRAFV600E mutation in archival paraffin-embedded tissues.

Clinicopathologic study of 85 colorectal serrated adenocarcinomas: further insights into the full recognition of a new subset of colorectal carcinoma.

Colorectal serrated adenocarcinoma represents a subtype of colorectal carcinoma that originates from serrated adenomas. Previous studies have suggested a more aggressive course, but this has not been verified. The aim of this work was to test the diagnostic reproducibility of previously proposed histologic criteria for serrated adenocarcinoma and to analyze the clinicopathologic features and outcome that would warrant its recognition as a new subtype of colorectal cancer. Nine hundred twenty-seven consecutive colorectal cancer specimens were used to search for cases fulfilling the criteria of serrated adenocarcinoma and matched controls. Clinicopathologic findings of 85 serrated adenocarcinomas were compared with a matched control group of conventional cancers. Serrated adenocarcinomas were encountered in 9.1% (n = 85) of cases. Residual serrated adenoma was present in 44 (51.7%). Absence of residual adenoma did not have any influence on the parameters studied. Interobserver variation between 2 Spanish and a Finnish pathologist showed moderate agreement (κ = .5873). Compared with their matched controls, serrated adenocarcinomas were more often accompanied by synchronous residual serrated adenomas (P < .0001), remote serrated adenomas (P = .0035), and serrated adenocarcinomas or cancers representing partial features of these tumors (P = .002). They had a less favorable 5-year survival than conventional cancers (P = .048 Breslow, Kaplan-Meier), and left-sided ones had the worst prognosis (P = .001). Serrated adenocarcinoma is an identifiable subset of colorectal cancer; and the histopathologic differences, in addition to its less favorable prognosis, may justify its recognition as a distinct subset of colorectal cancer warranting the search for specific clinical management strategies.

Human papillomavirus genotyping in histological sections of precursor lesions of cervical carcinoma: its role as a possible adjunct for the evaluation of the oncogenic potential of specific human papillomavirus genotypes - a study in a coastal region of southeastern Spain.

BACKGROUND: Human papillomavirus (HPV) genotyping is usually performed on cytological specimens with the aim of discerning between high- and low-risk genotypes. METHODS: Paraffin-embedded sections (n = 241) comprising 16 normal/benign (N/B) cervical sections, 72 low-grade squamous intraepithelial lesions (LSIL), 133 high-grade SIL (HSIL), 6 invasive carcinomas (cervical cancer), and 14 atypical immature metaplasias (AIMs) were DNA extracted and HPV genotyped. RESULTS: The most frequent HPV genotypes found were 16 and 58. HPV16 was detected in 0% N/B, 18.1% LSIL, 42.9% HSIL (p < 0.001), 50% carcinoma, and 35.7% AIM, whilst HPV58 was detected in 25.0, 20.8, 16.5, 0 and 35.7% of these lesions, respectively. DISCUSSION: The high prevalence of HPV58 and the low prevalence of HPV18 suggest the limited effectiveness of HPV vaccination in southeast Spain (prevention of 45.1% HSILs). The HPV genotype distribution profile in AIM suggests that these lesions are more similar to LSIL than HSIL pointing to a low risk of progression to cervical cancer. These results reinforce the necessity of assessing the specific genotype rather than distinguishing between high- or low-risk HPV. The use of histological section instead of cytological specimens for specific HPV genotyping would be very useful in order to ascertain the oncogenic potential of each of the genotypes found in a given area.

Expression profiles of ProEx C and Ki67 in squamous cell carcinoma in situ of the skin and their relationship with human papillomavirus genotypes.

BACKGROUND: ProExC is a new marker for identification of precursor lesions of cervical carcinoma. Its utility in noncervical squamous cell carcinoma in situ (SCCIS) such as Bowen's disease (BD) and actinic keratosis (AK) where human papillomavirus (HPV) plays a role has not been elucidated. Our aim was to ascertain the immunohistochemical features and clinical utility of ProExC in SCCIS of the skin. METHODS: HPV presence was tested in SCCIS (38 BD and 7 AK) using GP5+/6+ and Short PCR fragment (SPF) primers and subsequently genotyped. Histopathologic sections were stained for ProExC and Ki67. A set of non-neoplastic skin proliferative lesions were included for immunohistochemical evaluation [14 psoriasis (PS) and 6 psoriasiform dermatitis (PSD)]. RESULTS: HPV was detected in 18.9% BD. ProExC and Ki67 in the whole epidermis thickness was observed in 86.5 and 37.1% BD, respectively (p < 0.0001). ProExC and Ki67 were restricted to the lower third of the epidermis in PS and PSD. CONCLUSIONS: ProExC expression is not associated with HPV in SCCIS of the skin. Proliferating cells are better delineated in SCCIS by ProExC which may be useful to assess the extent of these lesions. Different immunohistochemical profiles seen in neoplasic and non-neoplastic skin lesions suggest diverse alteration of cell-cycle kinetics.

Effect of human papillomavirus on cell cycle-related proteins p16, Ki-67, Cyclin D1, p53, and ProEx C in precursor lesions of cervical carcinoma: a tissue microarray study.

In-depth study of cell cycle proteins and human papillomavirus (HPV) genotyping can provide useful information about the malignant potential of precursor lesions of cervical carcinoma (CC). Immunostaining of cell cycle-related proteins (p16, cyclin D1, Ki-67, p53, and ProEx C) was evaluated using tissue microarrays, and HPV genotypes were identified in 144 cervical tissue specimens encompassing normal or benign epithelial lesions, low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), and CC. In addition, 14 cases with atypical immature metaplasia (AIM) were included to compare their immunohistochemical features with those of well-established precursor lesions. Expression of p16, Ki-67, and ProEx C was most associated with the severity of dysplasia. Positive expression of p16, Ki-67, and ProEx C and negative expression of p53 seem to be related to HPV-16 infection. AIM cases show an immunohistochemical pattern more similar to LSIL than to HSIL. Immunohistochemical assessment of cell cycle proteins may help to distinguish normal and benign conditions of the cervix from precursor lesions of CC.

Genotype distribution of human papillomavirus (HPV) and co-infections in cervical cytologic specimens from two outpatient gynecological clinics in a region of southeast Spain.

BACKGROUND: Human Papillomavirus (HPV) genotype distribution and co-infection occurrence was studied in cervical cytologic specimens from Murcia Region, (southeast Spain), to obtain information regarding the possible effect of the ongoing vaccination campaign against HPV16 and HPV18. METHODS: A total of 458 cytologic specimens were obtained from two outpatient gynecological clinics. These included 288 normal benign (N/B) specimens, 56 atypical squamous cell of undetermined significance (ASC-US), 75 low-grade squamous intraepithelial lesions (LSIL) and 39 high-grade squamous intraepithelial lesions (HSIL). HPV genotyping was performed using PCR and tube array hybridization. RESULTS: The most frequent genotype found was HPV16 (14.9% in N/B; 17.9% in ASC-US; 29.3% in LSIL and 33.3% HSIL). Distribution of other genotypes was heavily dependent on the cytologic diagnoses. Co-infections were found in 15.3% of N/B, 10.7% of ASC-US, 48% of LSIL and 25.6% of HSIL cases (significantly different at p < 0.001). Strikingly, in N/B diagnoses, genotypes from A5 species were found as coinfecting in all cases. Genotypes from A7 or A9 species appeared in co-infections in 56.5% and 54% respectively whereas genotypes from A6 species appeared in 25.1% of cases. CONCLUSION: HPV vaccination might prevent 34.6% and 35.8% of LSIL and HSIL, respectively. Co-infection rate is dependent on both cytologic diagnosis and HPV genotype. Moreover, genotypes belonging to A5, A7 and A9 species are more often found as co-infections than genotype pertaining to A6 species. This suggests that phylogenetically related genotypes might have in common similar grades of dependency for cervical epithelium colonization.