Abstract
Simple SummaryDespite the availability of screening programs, oral cancer is often diagnosed due to the lack of effective biomarkers. Therefore, the identification of new effective diagnostic and late prognostic biomarkers is of fundamental importance for the management of this tumor type. In our previous computational study, we have identified a set of microRNAs (miRNAs) significantly dysregulated in oral cancer and with a potential diagnostic and prognostic significance for oral cancer patients. Starting from our preliminary bioinformatics results, the aim of the present study was to validate the diagnostic potential of four selected miRNAs, hsa-miR-133a-3p, hsa-miR-375-3p, hsa-miR-503-5p and hsa-miR-196a-5p, in liquid biopsy samples obtained from oral cancer patients and healthy donors. For this purpose, the expression levels of the selected miRNAs were determined in plasma samples by using specific miRNA probes and droplet digital PCR (ddPCR). The ddPCR results showed that the hsa-miR-133a-3p and hsa-miR-375-3p were significantly down-regulated in oral cancer and their evaluation in liquid biopsy samples can predict the risk of oral cancer development with high sensitivity and specificity. Finally, the computational analysis of miRNA expression and clinical-pathological features of patients allowed us to establish the functional role and prognostic significance of the two validated miRNAs.Despite the availability of screening programs, oral cancer deaths are increasing due to the lack of diagnostic biomarkers leading to late diagnosis and a poor prognosis. Therefore, there is an urgent need to discover novel effective biomarkers for this tumor. On these bases, the aim of this study was to validate the diagnostic potential of microRNAs (miRNAs) through the analysis of liquid biopsy samples obtained from ten oral cancer patients and ten healthy controls. The expression of four selected miRNAs was evaluated by using droplet digital PCR (ddPCR) in a pilot cohort of ten oral cancer patients and ten healthy donors. Bioinformatics analyses were performed to assess the functional role of these miRNAs. The expression levels of the predicted down-regulated hsa-miR-133a-3p and hsa-miR-375-3p were significantly reduced in oral cancer patients compared to normal individuals while no significant results were obtained for the up-regulated hsa-miR-503-5p and hsa-miR-196a-5p. ROC analysis confirmed the high sensitivity and specificity of hsa-miR-375-3p and hsa-miR-133a-3p. Therefore, both miRNAs are significantly down-regulated in cancer patients and can be used as biomarkers for the early diagnosis of oral cancer. The analysis of circulating miRNAs in a larger series of patients is mandatory to confirm the results obtained in this pilot study.
Highlights
Oral cancer represents one of the most prevalent tumors of the digestive tract, accounting for more than 350,000 novel diagnoses and 177,000 deaths every year [1]
A blinded pilot case series of ten oral cancer patients and ten healthy donors was included in the study in order to validate the prognostic value of hsa-miR-133a-3p, hsa-miR-375-3p, hsa-miR-196a-5p and hsa-miR-503-5p previously identified as key miRNAs involved in oral cancer development
The ten oral cancer patients enrolled in the study had a diagnosis of oral squamous cell carcinoma (OSCC)
Summary
Oral cancer represents one of the most prevalent tumors of the digestive tract, accounting for more than 350,000 novel diagnoses and 177,000 deaths every year [1]. As described for other tumors, several studies have demonstrated that the development of oral cancer is strictly related to numerous modifiable factors, including environmental and lifestyle risk factors, underlying the neoplastic transformation of cells [5,6]. Other causative agents of oral cancer are some microbial infections mainly due to Epstein Barr virus (EBV), human papillomaviruses (HPVs) or Candida albicans infection responsible for the activation of prooncogenic stimuli [9,10,11] In this context, recent reviews of the literature have highlighted a double role of microbial infections and microbiota composition in preventing or favoring the onset of tumors, including that of the oral cavity [12,13]
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