Squamous cell carcinoma research paper

The process of freeze-thawing or storage time has been investigated for its effect on detectable concentration of blood proteins. A recent study investigated the effects of freeze-thaw by comparing concentrations of inflammatory proteins, of which 9 overlap with our panel No significant differences were found between never-frozen and at least one freeze-thaw cycle. For a blood-based diagnostic test in clinical settings, immediate processing of fresh samples is convenient. However, to be cost-effective in clinical settings, MSD assay should analyze 80 collected samples.

Thus, subjecting samples to at least one freeze-thaw cycle may be unavoidable.

Original Research ARTICLE

Nevertheless, if the plasma biomarkers in this study are validated as OSCC specific, more economical clinical platforms, such as ELISA, can be designed to test on fresh-blood samples in clinical settings. The most intriguing observation from this study is the significant low level of these biomarkers in OSCC, compared to normal samples. Although this was unexpected, OSCC is known to be an immune inhibiting disease; therefore, the observed low-level expression may reflect this.

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Another possible explanation is that tumor-tissue associated inflammatory-related proteins are present at higher concentration at the tumor tissues than in the blood. Further investigation of the expression of these targets at the tumor tissue, unaffected tissues, and correlated to the circulating levels may shed light of its underlying mechanism.

However, given the multifaceted nature of these biomarkers, in which expression is affected by a variety of signaling pathways, the underlying biological explanation of the observed expression requires further experiments which are beyond the scope of this study.

Nevertheless, with the observed blood level we were able to estimate the performance in classifying OSCC which achieved high sensitivity and specificity. IL2 is predominantly produced by antigen-stimulated T cells, NK cells, and activated dendritic cells. MIF is a pro-inflammatory cytokine constitutively expressed and readily to be secreted by activated immune cells promoting cell proliferation and angiogenic activities, facilitating detection of antigens, and production of other inflammatory cytokines Therefore, the elevated level of IL1Ra in circulation may indicate the presence of inflammatory effects of IL1 in tumor tissues which trigger the IL1Ra to counterbalance the signaling pathways activated by IL1.

This suggests expression of IL1Ra plays a role in demoting the progression of tumor Several studies have demonstrated the expression of IL1Ra to be positively correlated with progression and lymph node metastasis 47 — 49 , inhibit IL-1 mediated prostate cancer regression 50 , and increased growth rate of glioblastoma cells These results may be suggesting that an increase in IL1Ra was to reduce the tumor-mediated production of IL1 52 and could propose value in assessing disease severity.

To explore the correlation among the candidate biomarkers using network visualization, we have observed interesting reverse relationships between biomarkers between OSCC and normal BCGP samples. This reflects previous reports demonstrating the production of MIF in presence of growth factors and inducing tumor growth 53 , In addition, the biomarkers among OSCC clustered to more subgroups suggesting biological difference within OSCC; although we did not have significant differences between clustered groups in regards to demographics, tumor clinical-pathological characteristics, or outcome.

The limitations of the study should be considered. First, this is not a large-scale mass spectrometry or a microarray study to comprehensively interrogate the complex plasma proteome. Therefore, biological sources of variability in observed expression such as protein isoforms, or pre- or post-transcriptional modifications could not be identified.

Instead, we wanted to apply a clinically translational platform to investigate the clinical value of immune-related biomarkers derived from easily accessible biosamples. Second, the retrospective nature of this study limits our full control over pre-analytical processing parameters, such as centrifugation time and speed, between laboratories, However, to our knowledge, there have been few reports of how centrifugation speed would significantly affect the detectable concentration of these proteins.

Tobacco and oral squamous cell carcinoma: A review of carcinogenic pathways

A few studies may even suggest that plasma-derived proteins are relatively robust to various sample processing methods 56 , In regards to study population, the normal matched samples from the BCGP are the best available samples that are most representative of the general non-OSCC population with comprehensive data collections on demographics, smoking and no known any cancer history with follow-up.

However, we do not have detailed medical information on whether there is presence of oral premalignant diseases, autoimmune diseases, and use of immunomodulators or other related conditions which could put this population at an increased risk of developing malignancies, which in turn may contribute to observed biomarker alterations. Therefore, it is worth to note that our OSCC population may not be generalized. Lastly, the observed aberrantly expressed protein may be due to changes in metabolic states or other physiological states that could not be captured in this study.

This limitation applies to all blood biomarker studies due to the varying genetic and non-genetic explanations, e. Future work is warranted to determine mechanisms by which most of these identified biomarkers are under-expressed in OSCC compared to normal. Other future directions may include a validation study with samples collected from different institutes with the determination of the best methods e.

ECL and cut-offs for various targets identified from this study. In addition, studies to investigate the temporal levels of these markers by repeating measurements before, post-treatment, and at time of local-regional recurrences or years into disease-free follow-up are of importance and can help to determine the value of these biomarkers in early identification of local and regional recurrence during the follow up. With further validation in larger sized cohort including paired blood samples collected over the course of disease management, the set of biomarkers has potential to assist in early detection of OSCC.

HK and CP contributed to the conception and design of the study. KL and NL performed the statistical analysis. KL wrote first draft of the manuscript. All authors interpreted and critically revised the manuscript to its final form. All authors gave final approval and agree to be accountable for all aspects of the work.

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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