Expression of microRNA-21 in saliva and tumor tissue of patients with oral squamous cell carcinoma: a predictor of cervical lymph node metastasis.

OBJECTIVE: Evaluation of diagnostic accuracy of microRNA-21 (miR-21) in saliva and tumor tissue for presurgical assessment of lymph node metastasis in patients with oral squamous cell carcinoma (OSCC). STUDY DESIGN: Unstimulated whole saliva and tumor tissue was obtained from clinically suspected patients with OSCC. A total of 130 patients diagnosed with OSCC were included as study participants. The assessment of cervical lymph node metastasis was done before surgery using imaging scans and post surgically confirmed by histopathologic examination of excised lymph nodes. miR-21 expression was evaluated using real-time polymerase chain reaction. The data was statistically analyzed for correlation analysis, cutoff values, sensitivity, and specificity. The κ statistic was applied to assess the degree of agreement between the lymph node metastasis and miR-21 expression. RESULTS: miR-21 expression showed a statistically significant correlation with cervical lymph node metastasis with a diagnostic accuracy of 65% to 71.54% in saliva and 69% to 81.54% in tumor tissue. Very good agreement was observed between tumor tissue miR-21-3p and cervical lymph node metastasis with a specificity of 80.60% and a sensitivity of 82.40%. CONCLUSIONS: miR-21 expression in saliva and tumor tissue of patients with OSCC showed high diagnostic accuracy for assessment of cervical lymph node metastasis. It can be used as an alternative for assessment of cervical lymph node metastasis before surgery.

miRNA for the assessment of lymph node metastasis in patients with oral squamous cell carcinoma: Systematic review and metanalysis.

BACKGROUND: miRNA is one of the advanced epigenetic molecular markers correlating with lymph node metastasis in patients with Oral squamous cell carcinoma (OSCC). Numerous published papers are showing correlation of miRNA with metastasis. There is a need to analyze and validate such correlation. METHOD: English language literature in major databases from the last 20 years was searched using controlled vocabulary and keywords. Strict inclusion and exclusion criteria were followed for selection of studies. The quality assessment was done as per the QUADAS tool 2 by three independent reviewers. The metanalysis was performed by using random effect model. Standardized mean difference (SMD) was considered as the effect measure. Statistical software used was STATA version 13.1. RESULTS: With all inclusion and exclusion criteria, eight studies could qualify for metanalysis. The pooled estimate is found to be 0.13 (-0.35, 0.62), P = .585, which is statistically not significant. This indicates that there is a no significant difference in the fold change between metastasis and no metastasis groups. P-value of chi-square statistic for heterogeneity is <.001 (significant), and I-squared statistic is 87.2%, which indicates that heterogeneity is present to a considerable extent. Egger's test shows there is no publication bias involved (P = .819). CONCLUSION: The metanalysis showed no significant difference in the fold change of miRNA expression between metastasis and non-metastasis OSCC patients. Future studies can be directed to eliminate the heterogeneity among the studies noted in this analysis to confirm the role of miRNA for assessment of regional metastasis with special focus on tongue squamous cell carcinoma.

Assessment of cervical lymph node metastasis based on total RNA from saliva and tumor tissue in patients with oral squamous cell carcinoma: An observational study.

BACKGROUND: In case of oral squamous cell carcinoma (OSCC) most patients die within first 2 years due to metastasis. To overcome the limitations and drawbacks of the present available methods of assessment of lymph nodes metastasis, the search for alternative method is needed. AIM: The aim of the study is to evaluate the sensitivity, specificity and diagnostic accuracy of salivary and tumor tissue RNA for assessment of lymph node metastasis in patients with OSCC. METHODOLOGY: Patients histologically diagnosed with OSCC were included as participants. The unstimulated saliva and tumor tissue were collected and stored at deep freeze before surgical therapy. The pretreatment lymph node metastasis assessment was done by radioimaging investigation. The posttreatment histopathological status of cervical lymph nodes was noted. The RNA was isolated and quantified from stored saliva sample and tumor tissue. The collected data were statistically analyzed for specificity and sensitivity and significance. RESULTS: The area under curve for salivary RNA level is 0.647 and for tumor tissue RNA level is 0.628 with moderate predictability at 95% confidence interval. It was observed that the sensitivity was 63.50% and 71.40% and specificity was 62.70% and 58.80% for saliva and tumor tissue respectively with diagnostic accuracy of 63%-65%. The Kappa statistics showed moderate degree of agreement with high statistical significance (P ≤ 0.05). CONCLUSION: Saliva and tumor tissue RNA can be a good marker for pretreatment assessment of lymph node metastasis in patients with OSCC. Although the diagnostic accuracy which range from 63% to 65%, further characterization and study of specific mRNA, siRNA and miRNA may come out with high diagnostic accuracy.

Comparative assessment of salivary cotinine level and psychological dependence among tobacco users.

BACKGROUND: The aim of the study was to assess the correlation between salivary cotinine level and psychological dependence measured through Fagerstrom test for nicotine dependence (FTND) questionnaire among tobacco users. MATERIALS AND METHODS: This was a cross-sectional study, conducted on tobacco users. Participants with the present habit of tobacco chewing and smoking above the age of 16 years were included in the study. A standard questionnaire form of FTND revised version for smoking and smokeless form of tobacco were given to each participant. Each participant was asked to answer the questions as per their experience of tobacco consumption and calculate the total point score or FTND score. Salivary cotinine level assessment was done using commercial available NicAlert kit. RESULTS: When salivary cotinine level was correlated with different variables of both groups, it was observed that weak correlation between salivary cotinine level and FTND scoring in smokers group (r = 0.083) and also in smokeless group (r = 0.081). When two groups were compared for salivary cotinine level, statistically significant difference (P = 0.021) was observed, with smokeless group showing high level of salivary cotinine level as compared to smokers group. CONCLUSION: Salivary cotinine and psychological dependence through FTND scoring are not strongly correlating with each other. This indicates that dependence over tobacco is a separate phenomenon and cannot be assessed by salivary cotinine level. It is well accepted that salivary cotinine level is influenced by age of individual, duration of habit, and type of tobacco consumption.

Cytological approach for diagnosis of non-healing oroantral fistula associated with candidiasis.

Oroantral fistula (OAF) and oral candidiasis are common to come across as separate individual lesions. However when candida organisms infect maxillary sinus through OAF then diagnosis should not be limited to clinical diagnosis only. In such situation role of cytological examination can prove to be fruitful. A female with chronic long standing OAF, not responding to conventional treatment approach is reported. On incisional biopsy, the case was diagnosed as chronic maxillary sinusitis with OAF. However patient did not respond to any treatment approach and later presented with a more progressive lesion involving maxillary sinus. A cytosmear stained with periodic acid Schiff stain, revealed the presence of numerous candidal hyphae. Finally, case was diagnosed as OAF with a superadded candidal infection. Patient responded well to antifungal treatment followed by reclosure of OAF. We should not neglect a simple cytological examination which may prevent wrong diagnosis and wrong treatment.

Clinicopathological prognostic implicators of oral squamous cell carcinoma: need to understand and revise.

In spite of the vast amount of research and the advances, oral squamous cell carcinoma implies quite significant mortality and morbidity rates. This has motivated the search of factors with prognostic relevance. A web based search was initiated for all published articles by using Medline/PubMed, Google Scholar with key words such as prognosis, survival rate, risk factors associated with oral squamous cell carcinoma, prognosis of oral squamous cell carcinoma. The search was restricted to articles published in English language with no restriction to date of publication. This review was focused on clinical, pathological and molecular factors associated with survival and prognosis of oral squamous cell carcinoma patients. Most articles had described one or two parameters related to prognosis. Considering the biological behavior and nature of cancer, all the parameters were interrelated and so could not predict the prognosis independently. Consideration of all the parameters was required to assess the prognosis. We hypothesize the use of combination of clinical and pathological indicators together to assess the prognosis. The care givers can assess the prognosis in a more better and definitive way by using prognosis assessment sheet.

Crystal violet stain as a selective stain for the assessment of mitotic figures in oral epithelial dysplasia and oral squamous cell carcinoma.

BACKGROUND: Assessment of mitotic figures (MFs) is routinely practiced as prognostic indicator in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC), but identification of MFs poses a problem in terms of staining characteristics. AIM: To evaluate effectiveness of crystal violet stain for staining of MFs and its comparison with hematoxylin and eosin (H and E) stain. MATERIALS AND METHODS: Study sample includes archival tissues embedded in paraffin blocks diagnosed as OED (n = 30) and OSCC (n = 30). The control group comprised of tissue specimen from oral mucosa of healthy volunteers (n = 30). Two serial sections of each tissue specimen were stained separately with H and E stain and 1% crystal violet stain. The stained sections were observed under microscope for identification and counting of MFs. Data obtained was statistically analyzed by using the Man-Whitney U test. RESULTS: A significant increase in number of MFs was observed in OED and OSCC in comparison with normal oral mucosa. There was a highly significant increase in number of MFs in crystal violet stained tissue sections when compared with H and E stain. Metaphase is the most commonly observed phase of mitosis in crystal violet stain when compared with H and E stain for all three groups. CONCLUSION: Crystal violet stain can be considered as selective stain for mitotic figures.