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Short-fragment PCR assay for highly sensitive broad-spectrum detection of human papillomaviruses in laryngeal squamous cell carcinoma and normal mucosa: clinico-pathological evaluation.

ABSTRACT: The aim of the study was to determine the prevalence and genotypes of HPV infection in laryngeal cancer specimens, normal mucosa obtained from the surgical margin and laryngeal nodules using a novel high sensitive and specific SPF(10) HPV DNA test, PCR/DEIA method and INNO-LiPA genotyping assay. The correlation between HPV presence and clinico-pathological features was analyzed. Tissue samples were collected from 93 primary laryngeal squamous cell carcinoma (LSCC), 49 specimens of normal mucosa and from 22 specimens of laryngeal nodules serving as control group. HPV DNA was amplified by the short PCR fragment (SPF(10)) primer set using HPV DNA enzyme immunoassay (DNA/DEIA) method and INNO-LiPA HPV genotyping assay. Human papillomavirus was detected in 33 (35.5%) of the 93 samples from LSCC, in 4 (8.2%) of 49 samples of the normal mucosa and it was not detected in any of the sample from the control group. Twenty-eight of 33 (81.8%) were positive for HPV-16, 6 of 33 (18.2%) were positive for HPV-18 and 5 of 33 (15.1%) were positive for HPV-33. Multiple infection was found in 5 of 33 (15.1%); 3 samples were positive for HPV-16 and HPV-33, 2 samples for HPV-16 and HPV-18. There was a statistically significant correlation between the presence of HPV in LSCC tumors and in control group samples and between the presence of HPV in the tumors and normal mucosa from the free surgical margin. The presence of HPV infection in 35.5% of the cases suggests a possible role in the etiology of laryngeal cancer and supports the role of high-risk types of HPV (16, 18 and 33) in LSCC. HPV infection is not likely to influence survival rates as an independent prognostic factor in patients with laryngeal cancer.

SUBMITTER: Morshed K

PROVIDER: S-EPMC2441493 | biostudies-other | 2008 Jul

REPOSITORIES: biostudies-other

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Project description:BackgroundTumorigenesis is a complex process of accumulated alteration in function of multiple genes and pathways. Wnt signalling pathway is involved in various differentiation events during embryonic development and is conserved in various species.ObjectiveA multicentre collaborative initiative is undertaken to study the occurrence, prognosis and molecular mechanism of HNSCC (Head and Neck Squamous Cell Carcinoma) which is highly prevalent in eastern parts of India. From a large cohort of HNSCC tissue repository, 67 cases were selected for multi-parametric investigation.Results67 cases showed stable ?-catenin expression. We have seen correlation, if any, of the transcription factor - ?-catenin, telomere maintenance and shelterin complex proteins - TRF2, Rap1 and hTert with respect to tumor differentiation and telomere dysfunction. Immunohistochemistry of ?-catenin protein showed stable and high expression in tumor when compared to stroma. MDSCC (Moderately Differentiated Squamous cell carcinoma) cases expressed nuclear expression of ?-catenin in invasive fronts and showed increased genomic instability. Higher frequency of Anaphase bridges was observed ranging from <3% in normal cut margin to 13% in WDSCC (Well differentiated squamous cell carcinoma) and 18% in MDSCC (Moderately differentiated Squamous cell carcinoma). There was significant decrease in telomere length in MDSCC (<4) when compared to the normal cut margin samples (<7). Quantitative Real Time-PCR confirmed a significant correlationship between stable ?-catenin expression and poor clinical and pathological outcome.ConclusionThe Stabilisation and accumulation of ?-catenin was significant and correlated well with de-differentiation process as well as prognosis and therapy outcome of the patients in the cohort. Expression status of molecular markers such as ?-catenin, hTert, TRF2 and RAP1 correlate significantly with the process of tumorigenesis and prognosis and may play a role in therapeutic management of Head and neck patients.

Project description:OBJECTIVE:Recently, a 24-h impedance was used to detect laryngopharyngeal reflux (LPR). However, not every case of LPR is pathological. Thus, pathological pharyngeal impedance values need to be clearly established to diagnose pathological LPR. The aim of our study was to establish pathological 24-h pharyngoesophageal impedance/pH values for the diagnosis of LPR. METHODS:The study was conducted in a tertiary care setting. A total of 30 patients who were referred to microlaryngoscopy for a laryngeal pathology that might be caused by LPR were included in this prospective study. All patients were off proton-pump inhibitor therapy. The 24-h pharyngoesophageal impedance-pH monitoring was performed 1 day before surgery. A biopsy of laryngeal tissue was obtained during microlaryngoscopy and was analyzed by immunohistochemistry to detect pepsin. The patients were divided into two groups: pepsin negative and pepsin positive (which indicated pathological LPR). The results of 24-h multichannel intraluminal impedance-dual-channel pH monitoring were compared between the groups. The number of LPR episodes in the pepsin-positive group was analyzed to establish a cutoff value for pathological LPR. RESULTS:There were 18 participants in the pepsin-negative group and 12 in the pepsin-positive group. The median total pharyngeal refluxes detected were two (0-5) in the pepsin-negative group and 14 (6-39) in the pepsin-positive group (P<0.001), although the groups were otherwise homogeneous. There was a statistically significant difference in the number of all types of refluxes between groups. Six or more pharyngeal refluxes were the cutoff for the presence of pepsin in the laryngeal mucosa and, thereby, for the diagnosis of relevant/pathological LPR. CONCLUSION:Six or more pharyngeal reflux episodes registered during the 24-h impedance/pH monitoring seem to be the cutoff for diagnosing pathological LPR. Therefore, it is possible to suggest establishing this value as the pathological impedance value indicating pathological LPR. These results must be interpreted with caution due to the small sample size.

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Short-fragment PCR assay for highly sensitive broad-spectrum detection of human papillomaviruses in laryngeal squamous cell carcinoma and normal mucosa: clinico-pathological evaluation.

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