Nasopharyngeal brushing: a convenient and feasible sampling method for nucleic acid‐based nasopharyngeal carcinoma research
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Tissue specimens for nasopharyngeal carcinoma (NPC) research are scarce because of sampling difficulties. Previous studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method for NPC diagnosis. The present study aimed to evaluate the feasibility of nasopharyngeal brushing in the acquisition of NPC nucleic acids for research.Nasopharyngeal brushing samples were acquired from 24 healthy individuals and 48 NPC patients. Tissues from 48 NPC and 18 nasopharyngitis patients were collected by endoscopic biopsy. The expression levels of tumor suppressor genes (TSGs) and Epstein-Barr virus (EBV)-encoded microRNAs as well as EBV DNA copy number were measured by quantitative polymerase chain reaction in both types of samples.Among six TSGs examined, the expression levels of two genes were significantly decreased in nasopharyngeal brushing and tissue samples from NPC patients as compared with those from healthy/nasopharyngitis individuals. Four EBV-encoded microRNAs, mir-bart1-5p, mir-bart5, mir-bart6-5p, and mir-bart17-5p, were significantly up-regulated in both NPC brushing and tissue samples compared with those from healthy/nasopharyngitis controls (P < 0.001). EBV DNA was significantly increased in both nasopharyngeal brushing samples (P < 0.001) and tissue samples (P < 0.001) from NPC patients in comparison with those from healthy controls.Nasopharyngeal brushing can obtain sufficient tumoral materials for the analysis of viral nucleic acid, including EBV-encoded microRNAs and EBV DNA. For the detection of TSG expression, nasopharyngeal brushings was feasible but inferior to tissue samples. This study confirms nasopharyngeal brushing as an applicable sampling method that can aid in nucleic acid-based NPC research.BackgroundElevated levels of antibodies against antigens in the Epstein–Barr virus (EBV) lytic phase are important predictive markers for nasopharyngeal carcinoma (NPC) risk. Several lifestyle factors, including smoking, have also been associated with NPC risk. We hypothesized that some specific lifestyle factors induce transformation of EBV from the latent to the lytic stage and contribute to NPC occurrence.
Lytic cycle
Epstein–Barr virus infection
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Epstein‐Barr virus (EBV) is known to be associated with two malignant diseases, nasopharyngeal carcinoma (NPC) and endemic Burkitt's lymphoma. In this study, the genomes of EBV in biopsy specimens from 4 NFC patients in Japan were analyzed using Southern blot hybridization. The NPC tissues of all examined cases contained rearranged EBV genomes whose Bam HI H fragments were larger than those of prototype EBV genomes. One of them had a Bam HI fragment containing contiguous sequences of Bam HI Y and H. A single‐sized EBV DNA terminus was observed in these NPC tissues, implying the evolution of the carcinoma from a single EBV‐infected cell.
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<i>Objectives:</i> To characterize the specific Epstein-Barr virus (EBV) polymorphisms from nasopharyngeal carcinoma (NPC) patients and healthy donors in Northern China, and to explore the relationship between the EBV genotypes and NPC. <i>Methods:</i> The genotypes of EBV strains were analyzed by the polymerase chain reaction and restriction fragment length polymorphism. <i>Results:</i> The predominant EBV type was A, C or F in both NPC and healthy individuals. The distributions of the EBV subtypes between NPC and healthy donors were not significantly different (p > 0.05). The frequency of type f strains in NPC was significantly lower in Northern China than in Southern China. <i>Conclusions:</i> No evidence of special EBV genotypes associated with NPC was found, suggesting that EBV strains derived from the NPC patients may reflect geographic distribution rather than being NPC restricted.
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Sera from 99 patients with nasopharyngeal carcinoma (NPC), 17 patients with diseases other than NPC and 24 healthy individuals were examined for their antibody activities to Epstein-Barr virus (EBV) DNase. Most of the sera from NPC patients showed high level of antibody activity even in the stage I of the disease. On the contrary, sera from healthy donors and patients with diseases other than NPC showed only very little or none of the activity. The results strongly suggest that the test for EBV DNase activity could be used for the early diagnosis of NPC.
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An Epstein-Barr virus (EBV) genome-positive epithelial hybrid cell line, NPC-KT, derived from the fusion of primary nasopharyngeal carcinoma cells with a human epithelial cell line of adenoid origin and a subline of EBV genome-positive Ramos cells, Ramos/NPC, converted after infection with NPC-KT EBV have been previously described (Takimoto et al., 1984; Takimoto et al., 1987). The NPC-KT cells produce virus (NPC virus) with both transforming and lytic properties. In this study, NPC-KT and Ramos/NPC cells were examined for the presence of the EBV receptor as measured by the capacity to absorb radio-labelled P3HR-1 and NPC viruses. It was determined that only P3HR-1 virus can attach to NPC-KT cells. Also, the relative concentration of NPC virus receptors on Ramos/NPC cells was found to be significantly reduced when compared to EBV genomenegative Ramos cells, whereas the relative concentration of receptors for P3HR-1 virus was similar to parental Ramos cells. The results suggest that there are differences at least in part of the receptors for P3HR-1 and NPC viruses.
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To observe the frequency of nasopharyngeal carcinoma (NPC) and its association with Epstein Barr Virus (EBV) infection.This study included consecutive cases of nasopharyngeal carcinoma, which were diagnosed in the Department of Pathology at the Aga Khan University Hospital, Karachi in the period of two years (1996-97).These tumors were initially evaluated on H&E stained sections. The tumors showing evidence of keratinization were excluded from the study. The Epstein Barr Virus was detected with the help of Polymerase chain reaction in formalin fixed, paraffin embedded tissue sections.During the study period, seventeen cases of nasopharyngeal carcinoma were diagnosed which comprised 0.3% of all malignant tumors. The age ranged from 5 years to 70 years with male to female ratio of 2.4:1. The NPC was more prevalent in adults (71%) as compared to children (29%) under 15 years. Six cases (35%) exhibited positive signal for Epstein Barr Virus.Nasopharyngeal carcinoma is an infrequent tumor. The prevalence of Epstein Barr virus infection in nasopharyngeal carcinoma is quite low as compared to other regions of the world.
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Immunoglobulin A
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Despite the fact that most adult humans worldwide are latently infected by the Epstein-Barr virus (EBV), only a very small percentage of them will develop an EBV-associated malignancy. We do not know whether this situation reflects the existence of more sensitive individuals or of particularly tumorigenic EBV strains. We postulated that if highly tumorigenic EBV strains did exist, they would be preferentially found in consistently EBV-associated tumors, such as nasopharyngeal carcinoma (NPC), and differ significantly from the strains present in other, non-pathological sites of the same patients. To test this hypothesis, we compared the BNLF1 gene of the EBV strains present in tumors and in “reservoir lymphocytes” of 6 NPC-bearing patients from Tunisia. Our results show that all of these patients were infected by more than 1 (and up to 7) EBV strains. Moreover, lymphocytes and tumor cells from the same individual were systematically infected by different viral strains. The origin and biological significance of these multistrain infections are discussed. © 2001 Wiley-Liss, Inc.
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Nasopharyngeal carcinoma (NPC) is an aggressive tumor with a complex etiology. Although Epstein-Barr virus (EBV) infection is known environmental factor for NPC development, the degree to which EBV naturally infects nasopharyngeal epithelium and the moment when and why the virus actively begins to affect cell transformation remains questionable. The aim of this study was to explore the association between LMP1 gene variability and potential contribution to NPC development. A systematic review was performed through searches of PubMed, Web of Science (WoS) and SCOPUS electronic databases. Additionally, meta-analysis of the difference in the frequency of seven LMP1 gene variants in NPC and control individuals was accomplished. The results from this study give a proof of concept for the association between 30 bp deletion (OR = 3.53, 95% CI = 1.48-8.43) and Xhol loss (OR = 14.17, 95% CI = 4.99-40.20) and NPC susceptibility when comparing biopsies from NPC and healthy individuals. Otherwise, 30 bp deletion from NPC biopsies could not distinguish NPC from EBV-associated non-NPC tumors (OR = 1.74, 95% CI = 0.81-3.75). However, B95-8, China1 and North Carolina variants were uncommon for NPC individuals. Much more efforts remains to be done to verify the biological significance of the differences observed, define so-called "high-risk" EBV variants and make it available for clinical application.
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Abstract It is important to know whether there are variants of Epstein‐Barr virus (EBV) with biological properties that are different from the prototype viruses that have been studied in detail, such as P3HR‐1 and B95‐8. We have studied an EBV isolate derived from a nasopharyngeal carcinoma (NPC) tumor, designated NPC‐EBV. We have examined the target B lymphocytes infected and growth‐transformed with NPC‐EBV as compared with two common EBV isolates, B95‐8 and AG876 EBV, for stage of maturation using antibodies to several immunoglobulin chains. Typing of the NPC‐EBV transformed lymphoblastoid cell lines revealed the predilection of the NPC‐EBV isolate to infect immature B lymphocytes. This was not the case for the B95‐8 and AG876 isolates. The reason for the predilection of NPC‐EBV for immature B lymphocytes remains to be explored further. However, these results may be important in understanding the pathophysiology of EBV‐associated diseases.
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