Role of miR-944/MMP10/AXL- axis in lymph node metastasis in tongue cancer.

Occult lymph-node metastasis is a crucial predictor of tongue cancer mortality, with an unmet need to understand the underlying mechanism. Our immunohistochemical and real-time PCR analysis of 208 tongue tumors show overexpression of Matrix Metalloproteinase, MMP10, in 86% of node-positive tongue tumors (n = 79; p < 0.00001). Additionally, global profiling for non-coding RNAs associated with node-positive tumors reveals that of the 11 significantly de-regulated miRNAs, miR-944 negatively regulates MMP10 by targeting its 3'-UTR. We demonstrate that proliferation, migration, and invasion of tongue cancer cells are suppressed by MMP10 knockdown or miR-944 overexpression. Further, we show that depletion of MMP10 prevents nodal metastases using an orthotopic tongue cancer mice model. In contrast, overexpression of MMP10 leads to opposite effects upregulating epithelial-mesenchymal-transition, mediated by a tyrosine kinase gene, AXL, to promote nodal and distant metastasis in vivo. Strikingly, AXL expression is essential and sufficient to mediate the functional consequence of MMP10 overexpression. Consistent with our findings, TCGA-HNSC data suggests overexpression of MMP10 or AXL positively correlates with poor survival of the patients. In conclusion, our results establish that the miR-944/MMP10/AXL- axis underlies lymph node metastases with potential therapeutic intervention and prediction of nodal metastases in tongue cancer patients.

Outcomes of a Telephone-Based Questionnaire for Follow-up of Patients Who Have Completed Curative-Intent Treatment for Oral Cancers.

Importance: Follow-up assessment of patients who had treatment for head and neck cancer is critical and an important part of the overall treatment program. During each visit to the hospital, patients are evaluated by a physician and may undergo additional tests. Because it has been observed that the symptoms mentioned by patients often guide the treating clinicians in identifying cancer recurrence, an appropriately constructed questionnaire can help clinicians determine which patients need further testing as a result of a recurrence and which patients can continue to be monitored remotely. Objective: To evaluate the role of a symptom-based telephone questionnaire in detecting recurrences in patients with radically treated oral cancer. Design, Setting, and Participants: This prospective diagnostic cohort study was conducted from October 1, 2018, to February 28, 2019. This study took place at Tata Memorial Centre, an apex referral cancer center in India. A total of 615 consecutive patients with oral cancer were screened, of whom 400 patients consented to be a part of the study. After completion of curative treatment, these patients were followed up for 2 months to 2 years. We excluded patients younger than 18 years or older than 80 years, those with Eastern Cooperative Oncology Group status greater than 2, and those who had already been diagnosed with recurrent disease. Interventions/Exposures: The patients were contacted 2 weeks before their clinic appointment, and a telephone interview was conducted using a predefined questionnaire. Based on patients' responses to these questions, their disease status was estimated. Subsequently, during the actual follow-up visit, a trained head and neck surgeon, who was blinded to the questionnaire result, examined them. Main Outcomes and Measures: The sensitivity and specificity of the telephone questionnaire were calculated by comparing its results with those of the trained head and neck surgeon after the clinical evaluation. Results: Of the 615 patients screened, 400 consented to be part of the study. Participants had a median (interquartile range) age of 49 (41-56) years, and 334 (83.5%) were men. Recurrence was noted in 20 patients (5.0%). The telephone-based questionnaire was found to have sensitivity of 90.0%, specificity of 74.2%, positive predictive value of 15.5%, and negative predictive value of 99.3%. Clinical examination values were 100.0%, 92.9%, 42.5%, and 100.0%, respectively. A total of 124 patients (31.0%) said that they would prefer such a telephone follow-up compared with an actual physical follow-up visit. Conclusions and Relevance: This diagnostic cohort study found that a symptom-based telephone questionnaire had good sensitivity and negative predictive value for detecting recurrences in patients with oral cancer on follow-up evaluation after completion of definitive treatment.

Notch pathway activation is essential for maintenance of stem-like cells in early tongue cancer.

BACKGROUND: Notch pathway plays a complex role depending on cellular contexts: promotes stem cell maintenance or induces terminal differentiation in potential cancer-initiating cells; acts as an oncogene in lymphocytes and mammary tissue or plays a growth-suppressive role in leukemia, liver, skin, and head and neck cancer. Here, we present a novel clinical and functional significance of NOTCH1 alterations in early stage tongue squamous cell carcinoma (TSCC). PATIENTS AND METHODS: We analyzed the Notch signaling pathway in 68 early stage TSCC primary tumor samples by whole exome and transcriptome sequencing, real-time PCR based copy number, expression, immuno-histochemical, followed by cell based biochemical and functional assays. RESULTS: We show, unlike TCGA HNSCC data set, NOTCH1 harbors significantly lower frequency of inactivating mutations (4%); is somatically amplified; and, overexpressed in 31% and 37% of early stage TSCC patients, respectively. HNSCC cell lines over expressing NOTCH1, when plated in the absence of attachment, are enriched in stem cell markers and form spheroids. Furthermore, we show that inhibition of NOTCH activation by gamma secretase inhibitor or shRNA mediated knockdown of NOTCH1 inhibits spheroid forming capacity, transformation, survival and migration of the HNSCC cells suggesting an oncogenic role of NOTCH1 in TSCC. Clinically, Notch pathway activation is higher in tumors of non-smokers compared to smokers (50% Vs 18%, respectively, P=0.026) and is also associated with greater nodal positivity compared to its non-activation (93% Vs 64%, respectively, P=0.029). CONCLUSION: We anticipate that these results could form the basis for therapeutic targeting of NOTCH1 in tongue cancer.