Immunohistochemical expression of Epstein-Barr virus-encoded latent membrane protein (LMP-1) in paraffin sections of EBV-associated nasopharyngeal carcinoma in Spanish patients

doi:10.1016/0964-1955(95)00093-3

European Journal of Cancer Part B: Oral Oncology

Volume 32, Issue 3, May 1996, Pages 163-168

European Journal of ...

https://doi.org/10.1016/0964-1955(95)00093-3 Get rights and content

Abstract

Nasopharyngeal carcinoma (NPC) paraffin samples, from Spanish patients, of distinct histological types, including squamous cell carcinoma (10 cases), nonkeratinising carcinoma (12 cases) and undifferentiated carcinoma (29 cases) were analysed for Epstein-Barr virus (EBV) detection and EBV-encoded latent membrane protein (LMP-1) expression using a sensitive nested-polymerase chain reaction with four oligonucleotide primers specific for EBV genome (EB-1, 2, 3, 4) and immunohistochemistry by means of CS1-4 pool monoclonal antibody. EBV genome was detected regardless of histological type in 100% of samples with sufficient DNA quality to permit viral diagnosis (50 out of 51 cases), supporting the previous view that all types of NPC are variants of an EBV-associated malignancy. However LMP-1, an EBV-encoded oncogenic protein, was detected in 40 out of 51 samples (78.4%) and LMP-1 immunohistochemical expression was not apparently influenced by histological type, primary or metastatic site, clinical stage, age or sex. This high percentage of detection of LMP-1 in our cases supports a role for EBV in the pathogenesis of different types of NPC, but the lack of constant expression of LMP-1 in NPC remains unclear and various reasons are postulated to explain the absence of this oncogenic protein in some EBV-associated NPCs.

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Cited by (31)

Patterns of aberrant DNA hypermethylation in nasopharyngeal carcinoma in Tunisian patients
2012, Clinica Chimica Acta
Citation Excerpt :
In our study EBV-DNA/RNA positivity and LMP1 expression were not influenced by histological type, clinical stage, age, and sex. These findings are in line with previous studies [34]. Moreover, we did not observe any significant correlation between the methylation status of individual genes and the expression of LMP1.
Aberrant methylation in the promoter of tumor-related genes is associated closely with epigenetically mediated gene silencing. The aim of the present study was to evaluate the methylation profile of Tunisian nasopharyngeal carcinoma (NPC) and to determine the clinicopathological features of tumors showing this epigenetic alteration.
Thirty-six archival NPC biopsies were investigated in comparison with 19 non-tumor nasopharyngeal tissue specimens. DNA methylation status of ten tumor-suppressor and related genes was analyzed by using methylation-specific PCR. The Epstein-Barr virus (EBV) presence was verified by PCR and in situ hybridization and the LMP1 oncoprotein expression was analyzed by immunohistochemistry. Findings were then correlated with clinicopathological variables (Patients' gender and age, tumor histological subtype and stage).
Hypermethylation frequencies of the investigated genes in NPC biopsies were 75% for RASSFIA, 58.3% for SHP1, 47.2% for DAPK, 33.3% for P16, 31% for RARβ2, 19.4% for GSTP1 and TIMP3, 11% for APC and CDH1, and 5.5% for MGMT. In non-tumor nasopharyngeal samples, hypermethylation was detected in lower frequencies in 6 genes (SHP 26.3%, P16 21%, RARβ2 21%, DAPK 15.8%, TIMP3 10.5%, and GSTP 5.3%).
Hypermethylation of RARβ2 promoter was more frequent in tumors with lymph node metastasis than those without metastasis (43.5% vs 0%, p = 0.03). Methylation of RASSF1A was more frequently detected in non-keratinizing NPC than in undifferentiated subtype (100% vs 66.7%; p = 0.05). A trend toward positive association was found between an increased number of methylated genes and LMP1 expression (p = 0.07). However, no significant association was found for the remaining variables.
This study indicates that hypermethylation of multiple genes is a common alteration in nasopharyngeal carcinomas in Tunisian patients and that this epigenetic change may play a role in the nasopharyngeal carcinogenesis.
P53 and nasopharyngeal carcinoma: A Malaysian study
2009, Pathology
Nasopharyngeal carcinoma (NPC) is a common malignancy among men in Malaysia. To determine the role of p53 in NPC, we screened for p53 mutations and evaluated the protein expression levels in samples from local patients with NPC.
Fifty-three formalin-fixed, paraffin-embedded nasopharyngeal carcinoma tissue blocks were chosen for this study. The presence of Epstein–Barr virus (EBV) was determined by in situ hybridisation using an EBER probe. p53 protein expression was detected using immunohistochemistry. Simultaneously, amplifications by PCR were performed for p53 exons 5 to 8, followed by mutation screening via single strand conformation polymorphism (SSCP). Sequencing of all the four exons was performed in five samples with mobility shift. To rule out false negative results by SSCP, 13 samples with p53 overexpression and five samples with low p53 expression were randomly selected and sequenced.
There was no mutation found in exons 5 to 8 in all the samples despite 46 (87%) of them having high p53 levels. EBV was detected in 51 (96%) out of 53 samples. There was no statistically significant association between p53 expression level and EBV presence
High-intensity staining for p53 by immunohistochemistry was common in our series of NPC tissue samples but was not associated with ‘hot spot’ mutations of exons 5–8 of the gene. We did not find a significant relationship between the expression level of p53 and presence of EBV. Our study confirms that mutation of the DNA-binding domain of p53 is rare in NPC.
Expression of p53 protein and tumor angiogenesis as prognostic factors in nasopharyngeal carcinoma patients
2002, Pathology Research and Practice
The objective of this study was to evaluate the possible prognostic significance of p53 protein overexpression and tumor angiogenesis (TA) in nasopharyngeal carcinoma (NPC) patients, together with other clinicopathological variables. Forty-two NPC patients were evaluated in relation to survival. Nuclear p53 overexpression in neoplastic and endothelial cells was detected by immunohistochemistry (IHC) with the monoclonal antibody DO-7 and the polyclonal antibody against factor VIII-related antigen, respectively. Thereafter, we evaluated p53 cases in order to determine their nuclear immunoreactivity from negative (–) to positive (+, ++, +++). In addition, microvessels were counted in the most active areas of tumor neovascularization or hotspots using an image computer analyzer (MicroImage^®).
A Cox multiple regression survival analysis was used to determine the best prognostic indicators in NPC patients. As a result, tumor microvessel count, considered as a continuous variable, was the most important independent prognostic indicator in relation to survival (p = 0.0273), with a relative risk of death of 2,4399 [95% confidence interval 1.1051; 5.3871] associated with the highest microvessel counts. Moreover, the only clinicopathological variable that demonstrated prognostic value in a Cox multiple regression survival analysis was histological type (p equals; 0.05). In addition, we did not observe any statistical association between intratumoral microvessel density (IMD), clinicopathological variables and p53 protein expression.
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PaperImmunohistochemical expression of Epstein-Barr virus-encoded latent membrane protein (LMP-1) in paraffin sections of EBV-associated nasopharyngeal carcinoma in Spanish patients

Abstract

Lancet

Cell

Lancet

Oral Oncol, Eur J Cancer

Oral Oncol, Eur J Cancer

Eur J Cancer

Nasopharyngeal carcinoma: the Epstein-Barr virus association

Semin Oncol

Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2

Epstein-Barr virus replication in oropharyngeal epithelial cells

N Engl J Med

Immunoglobulin A-induce shift of Epstein-Barr virus tissue tropism

Science