Downregulation of salivary miR-3928 as a potential biomarker in patients with oral squamous cell carcinoma and oral lichen planus.

OBJECTIVES: Recent studies highlighted the role of miR expressed in saliva as reliable diagnostic and prognostic tools in the long-term monitoring of cancer processes such as oral squamous carcinoma (OSCC). Based on a few previous studies, it seems the miR-3928 can be considered a master regulator in carcinogenesis, and it can be therapeutically exploited. This is the first study that compared oral potentially malignant disorder (OLP) and malignant (OSCC) lesions for miR-3928 expression. MATERIALS AND METHODS: In this cross-sectional study, saliva samples from 30 healthy control individuals, 30 patients with erosive/atrophic oral lichen planus, and 31 patients with OSCC were collected. The evaluation of miR-3928 expression by q-PCR and its correlation with clinicopathological indices were analyzed by Shapiro-Wilk, Kruskal-Wallis, Pearson's χ2 , and Mann-Whitney tests. The p-value less than .05 indicated statistically significant results. RESULTS: Based on nonparametric Kruskal-Wallis test results, there was a statistically significant difference between the ages of three study groups (p < .05). This test demonstrated a statistically significant difference between the average of miR-3928 expression in three study groups (p < .05). The result of the χ2  test showed a statistically significant difference in miR-3928 expression between patients with OLP (p = .01) and also patients with OSCC (p < .0001) in comparison to the control group. CONCLUSIONS: The salivary miR-3928 can play a tumor suppressive role in the pathobiology of OSCC, and it is significantly downregulated in patients. According to the potential tumor suppressive role of miR-3928 in the OSCC process, we can consider this microRNA as a biomarker for future early diagnosis, screening, and potential target therapy.

Association between Vitamin D Receptor Polymorphism and Susceptibility to Oral Lichen Planus and Oral Squamous Cell Carcinoma.

Introduction: Oral squamous cell carcinomas (OSCC) comprise 90-95% of oral cancers. Early diagnosis improved the survival rate of OSCC patients to 80-90%. Oral lichen planus (OLP) is a chorionic inflammatory disease with malignancy potential. The vitamin D receptor (VDR) plays a critical role in the pathogenesis of oral cancer. This study aimed to determine the association between VDR rs7975232 (Apa I) polymorphism and potential susceptibility to OLP and OSCC risks. Materials and Methods: In this prospective case-control study, a total of 120 blood samples were obtained from OSCC patients (n=29), OLP (n=50), and controls (n=40). VDR rs7975232 polymorphism was studied using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) method. Statistical analysis was performed with SPSS Version 23 software. Data were expressed as means ± standard deviation (SD). Age, sex, allelic frequency, and genotyping were compared using the chi-square test. A p-value of less than 0.05 was regarded as statistically significant. The disease risk was estimated by Odds ratio (OR) with a 95% confidence interval. Results: A significant age difference was observed between the controls and the OSCC group (p=0.001). A significant difference was observed in Aa and aa genotypes compared with AA between OSCCs and controls. Moreover, dominant (p<0.001), additive (p<0.001), and allelic (p=0.001) models were different between groups. Conclusion: There was a positive association between rs7975232 VDR polymorphism and susceptibility to OSCC. More experimental evidence must reveal the possible association between rs7975232 and the risk of OLP in a larger cohort.

Clinical value and potential circulating of miR-99a as tumor suppressor biomarker in serum of oral squamous cell carcinoma and erosive atrophic lichen planus.

OBJECTIVES: Oral squamous cell carcinoma (OSCC) is the most common type of oral neoplasms that consist of more over 90% of oral cancers. It was demonstrated that erosive atrophic oral lichen planus (OLP) has potential of malignancy transformation into OSCC. The microRNAs are non-coding regulator sequences involved in cancer process. The miR-99a involve in growth, proliferation, migration, invasion, and metastasis of tumor cells. Therefore, we evaluated miR-99a expression in serum of OSCC and erosive atrophic OLP patients in comparison to healthy control individuals to more investigate about level of miR-99a expression in potential premalignant disorder (erosive atrophic OLP) in comparison to malignant transformation form (OSCC). Gene ontology (GO) and pathway analyses were performed to better understand the importance of miR-99a in OSCC. MATERIALS AND METHODS: In this cross-sectional study, total 90 serum samples from OSCC patients (n = 30), erosive atrophic OLP (n = 30) and healthy control individuals (n = 30) were collected, and then evaluated for miR-99a expression by qPCR. Pathway analysis and protein-protein interaction were done using STRING (v: 12.0), and (GO) terms and related genes were extracted from the GO online search tool. The statistical analysis was evaluated by Kruskal Wallis, Chi-Square, Kruskal Wallis, Spearman and Mann-Whitney tests. The p-value less than 0.05 was considered statistically significant. RESULTS: miR-99a expression down regulated in OSCC in comparison to erosive atrophic OLP and control groups (p < 0.05). The miR-99a up regulated in grade I more than grades II and III (p < 0.05). We showed upregulation of miR-99a in early stage more than advanced stage (p < 0.05). Expression of miR-99a reduced accordance to the increasing of tumor size and lymph involvement levels (p < 0.05). The 165 determined targets were classified into three domains. The most significant enrichment in biological processes, cellular components, and molecular functions was in the cellular nitrogen compound biosynthetic process, cytosolic ribosome, and protein binding, respectively. CONCLUSIONS: We highlighted tumor suppressive role of miR-99a in OSCC patients. It seems that miR-99a can be considered a valuable biomarker for the early diagnosis of erosive atrophic OLP before transformation. CLINICAL RELEVANCE: Our results may help to better understand the prognostic factor for oral squamous cell carcinoma to evaluate survival and subsequent tumor development. And it may also help to understand the pathogenesis of OSCC.

Unveiling the Potential of Serum MiR-483-5p: A Promising Diagnostic and Prognostic Biomarker in OLP and OSCC Patients by In silico Analysis of Differential Gene Expression.

BACKGROUND: Oral squamous cell carcinoma (OSCC) and oral lichen planus (OLP) are two separate conditions affecting the mouth and result in varying clinical outcomes and levels of malignancy. Achieving early diagnosis and effective therapy planning requires the identification of reliable diagnostic biomarkers for these disorders. MicroRNAs (miRNAs) have recently received attention as powerful biomarkers for various illnesses, including cancer. In particular, miR-483-5p is a promising diagnostic and prognostic biomarker in various cancers. Therefore, this study aimed to investigate the role of serum miR-483-5p in the diagnosis and prognosis of OLP and OSCC patients by in silico analysis of differential gene expression. METHODS: GSE23558 and GSE52130 data sets were selected, and differential gene expression analysis was performed using microarray data from GSE52130 and GSE23558. The analysis focused on comparing OLP and OSCC samples with normal samples. The genes intersected through the differential gene expression analysis were then extracted to determine the overlapping genes among the upregulated or downregulated DEGs. The downregulated genes among the DEGs were subsequently imported into the miRWalk database to search for potential target genes of miRNA 483-5p that lacked validation. To gain insight into the biological pathways associated with the DEGs, we conducted pathway analysis utilizing tools, such as Enrichr. Additionally, the cellular components associated with these DEGs were investigated by analyzing the String database. On the other hand, blood serum samples were collected from 35 OSCC patients, 34 OLP patients, and 34 healthy volunteers. The expression level of miR-483-5p was determined using quantitative reverse transcription polymerase chain reaction (RT-qPCR). The Kruskal-Wallis test was utilized to investigate the considerable correlation. Moreover, this study explored the prognostic value of miR-483-5p through its association with clinicopathological parameters in OSCC patients. RESULTS: The results showed that serum expression of miR-483-5p was considerably higher in OSCC patients compared to OLP patients and healthy controls (p 0.0001) and that this difference was statistically significant. Furthermore, elevated miR-483-5p expression was associated with tumor size, lymph node metastasis, and stage of tumor nodal metastasis in OSCC patients (p 0.001, p 0.038, and p 0.0001, respectively). In silico analysis found 71 upregulated genes at the intersection of upregulated DEGs and 44 downregulated genes at the intersection of downregulated DEGs, offering insight into the potential underlying mechanisms of miR-483-5p's engagement in OSCC and OLP. The majority of these DEGs were found to be involved in autophagy pathways, but DEGs involved in the histidine metabolism pathway showed significant results. Most of these DEGs were located in the extracellular region. After screening for downregulated genes that were invalidated, miRNA 483-5p had 7 target genes. CONCLUSION: This study demonstrates the potential of serum miR-483-5p as a promising diagnostic and prognostic biomarker in OSCC and OLP patients. Its upregulation in OSCC patients and its association with advanced tumor stage and potential metastasis suggest the involvement of miR-483-5p in critical signaling pathways involved in cell proliferation, apoptosis, and cell cycle regulation, making it a reliable indicator of disease progression. Nevertheless, additional experimental studies are essential to validate these findings and establish a foundation for the advancement of targeted therapies and personalized treatment approaches.

The oncogenic role of NOTCH1 as biomarker in oral squamous cell carcinoma and oral lichen planus.

Background: Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer with heterogeneous molecular pathogenesis. Oral lichen planus (OLP) is demonstrated potentially can transfer to OSCC malignant lesions. Unfortunately, there are no definitive prognostic and predictive biomarkers for the clinical management of OSCC patients. The present research is the first study that compared an oral premalignant lesion such as OLP to malignant lesions like OSCC for NOTCH1 expression levels to better understand its oncogenic or tumor suppressive role. Materials and Methods: In this cross-sectional study, mRNA expression of NOTCH1 was evaluated by quantitative polymerase chain reaction in 65 tissue-embedded Paraffin-Block samples, including 32 OSCC and 33 OLP. Furthermore, we collected demographic information and pathological data, including tumor stage and grade. The association between NOTCH1 and GAPDH gene expressions was determined by Chi-squared, Spearman, and Mann-Whitney tests. A P < 0.05 was considered statistically significant for all statistical analyses. Results: Comparison of OSCC and OLP groups showed a statistically significant difference between the quantitative expression of the NOTCH1 gene (P < 0.001). Qualitative gene expression was divided into low expression and high expression. Both study groups demonstrated a statistically significant gene expression difference (P < 0.001). There was a statistically significant difference between age and NOTCH1 expression in the OLP group (P = 0.036). There was no correlation between NOTCH1 expression and age, gender, tumor grade, and stage. Conclusion: Since the OSCC is a malignant lesion and the OLP showed the possible nature of malignancy transformation, we can consider the NOTCH1 as a biomarker for the assessment of the tumorigenesis process with a definition of a standard threshold for potentially malignant lesions and malignant OSCC tumors.

RT-qPCR Analysis of LAMP3 (CD208) Gene Expression in Oral Lichen Planus and Oral Squamous Cell Carcinoma.

Background: Many new studies have been conducted on cellular proteins to use them as prognostic markers or in target therapy through determining the increase or decrease in their expression in the lichen planus and OSCC. LAMP3 protein is one of these proteins which has been recently considered. Thus, considering the unknown etiology of lichen planus, significance of their early diagnosis and treatment and lack of a suitable and final treatment for this disease and oral cancers, and preventing the progression of lichen planus, which can turn into OSCC, we decided to investigate the level of expression of this gene and its effect on the progression, study the connection between these two conditions and the probable factors contributing to their etiopathogenesis. Methods: In this study, ninety-four paraffin blocks tissue samples of patients were obtained together with their demographic documents. LAMP3 expression was measured RT-qPCR method. Results: The results show that there is not any significant difference between age and sex population of our study. in squamous cell carcinoma the amount of expression of LAMP3 was higher than lichen planus and healthy margin. Average LAMP3 Gene expression in grade III was higher than group grade I & II in which considering significant level of 5%, it is statistically significant. Conclusions: According to the findings of this study, it can be concluded that the expression of the LAMP3 gene in SCC lesions is higher than in healthy tissue. Hence, LAMP3 gene expression can be used as a diagnostic biomarker.

Salivary Antiviral and Antibacterial Properties in the Encounter of SARS-CoV-2.

Due to the high mortality rate of COVID-19 and its high variability and mutability, it is essential to know the body's defense mechanisms against this virus. Saliva has numerous functions, such as digestion, protection, and antimicrobial effects. Salivary diagnostic tests for many oral and systemic diseases will be available soon because saliva is a pool of biological markers. The most important antiviral and antibacterial compounds identified in saliva include lysozyme, lactoferrin (LF), mucins, cathelicidin, salivary secretory immunoglobulin (SIgA), chromogranin A, cathelicidin, salivary agglutinin (SAG) (gp340, DMBT1), α, ß defensins, cystatin, histatins, secretory leukocyte protease inhibitor (SLPI), heat shock protein (HSP), adrenomedullin and microRNA (miRNAs). Antimicrobial peptides (AMPs) in saliva could be used in the future as models for designing effective oral microbial antibiotics. The antiviral properties of the peptides in saliva may be one of the future treatments for the COVID-19 virus. In this review, we investigate compounds with antiviral and antibacterial properties in saliva and the importance of these compounds in saliva in exposure to the COVID-19 virus. Due to the transmission route of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) into the oral cavity in the lower and upper respiratory tract, studies of salivary antiviral properties in these patients are very important. Some of the antiviral effects of saliva, especially mucin, α, ß-defensins, IgA, IgG, IgM, lysozyme, SAG, SLPI, and histatins, may play a greater role in neutralizing or eliminating COVID-19.

Necroptosis has a crucial role in the development of chronic periodontitis.

Background and aim: Periodontitis is a non-communicable chronic inflammatory disease that affects the entire periodontium and its severe types cause irreparable destruction. The purpose of this study was to determine the type of cell death in chronic periodontitis (CP) with the expression of receptor-interacting protein kinase (RIPK) type1 and RIPK3 genes. Materials and methods: This cross-sectional study was carried out from September 2019 to 2020. The samples (38 participants) were divided into two groups: 20 recently diagnosed CP patients and 18 healthy individuals. Participants' data was collected in the periodontology Department, Dental school, Mashhad University of Medical Sciences and sent to the Immunology Lab for assessment of RIPK1 and RIPK3 expressions using quantitative real time-PCR. Results: The study sample consisted of 30 females (78.9%) and 8 males (21.1%) with a mean age of 34 ± 5 years. The expression of the genes of interest in CPs exhibited an opposite pattern. Although, RIPK3 gene expression was significantly greater in CP patients compared to the control group (P = 0.024), the expression of RIPK1 decreased (p < 0.001). Moreover, no significant correlation was observed between age and gender with these molecules in CPs. Conclusion: The RIPK3 selectively contributes to necroptosis, therefore, it seems that RIPK3-mediated necroptosis is involved in chronic periodontitis. RIPK1 also participates in necroptosis, but mostly in apoptosis. Therefore, necroptosis as an unprogrammed inflammatory cell death induced by pathogenic damages seems to be another mechanism complicated in periodontitis and could be used as a novel target for CP therapy.

Matrix Metalloproteinase-3 but Not Matrix Metalloproteinase-9, Implicated in the Manifestation of Chronic Periodontitis.

Background: Periodontal disease is an inflammatory condition affecting the tooth's supporting tissues, resulting in gradual loss of periodontal ligament (PDL), alveolar bone, and gum resorption. Neutrophil and monocyte/macrophage, destructive proteases like matrix metalloproteinase (MMP)-3 and MMP-9 play pivotal roles in such lesions in periodontitis. Therefore, this study aims to compare the level of MMP-3 and MMP-9 gene expression in patients with or without periodontitis in an Iranian population. Methods: This cross-sectional study was carried out on 22 chronic periodontitis patients and 17 healthy control subjects referred to the department of periodontology, Mashhad Dental School. In both groups, the gingival tissue was removed during surgery and transferred to the Molecular Biology Laboratory for MMP-3 and MMP-9 gene expression evaluation. The qRT-PCR, TaqMan method was used for gene expression assessments. Results: The average age of periodontitis patients was 33± 5 years, and in controls, 34.7± 6 with no significant differences. The mean MMP-3 expression in periodontitis patients was 146.67±38.7, and in controls, 63.4±9.1. The difference was statistically significant (P=0.04). The mean expression of MMP-9 in periodontitis patients and controls were 103.8± 21.66 and 87.57± 16.05, respectively. Although the target gene expression in patients was higher, the difference was insignificant. Furthermore, there was not any significant correlation between age or gender with the expression of MMP3 or MMP9. Conclusion: The study demonstrated that the MMP3 seems to have a destructive impact on the gingival tissue in chronic periodontitis, but not MMP9.

Associations between RORγt and T-bet Expressions, clinicopathological indices and survival rate in oral Squamous cell carcinoma patients.

BACKGROUND: Oral cancers are the sixth most common cancers around the world. According to the pivotal role of immune cells in the pathogenesis of oral squamous cell carcinoma (OSCC), as the frequent form of malignant epithelial neoplasm in the oral cavity, we investigated the association between the expression of RORγt and T-bet genes as two transcription factors, clinicopathologic indices, and survival rate. METHODS AND MATERIALS: Forty-two OSCC paraffin embded-blocks tissue samples and their surgical healthy margins (as a control group) were collected. Demographic information like age and gender, and medical history including tumor stage/grade, and following-up time were registered. The RORγt and T-bet expression were assessed by qPCR. The overall survival (OS) and disease free survival (DFS) were analyzed by SPSS V.23 software. RESULTS: The expression of RORγt and T-bet genes in OSCC patients were significantly higher than in surgical healthy margins (P < 0.001). Both expression demonstrated a significant difference between surgical healthy margins and tumor tissues related to gender and clinicopathological indices including stage and grade (P < 0.05). The expression of both genes in stage I patients was significant compared to stage IV (P < 0.05). The relation between expressions, OS, and DFS with clinical stage and histological grade of tumors was not statistically significant (P > 0.05). CONCLUSION: Overexpression of RORγt and T-bet in OSCC patients with higher grade and stage in compare to surgical healthy margin highlighted their critical role in OSCC pathogenesis including oral epithelial cell differentiation, tumorigenesis process, and malignant transformation. Moreover, both mentioned genes can apply as prognostic biomarkers in OSCC patients. We suggest surgical healthy margin be considered as valuable biological area.

Evaluation of CD4+ tumor-infiltrating lymphocyte association with some clinicopathological indices of oral squamous cell carcinoma.

Background: The delayed diagnosis of oral squamous cell carcinoma (OSCC) affects therapeutic and prognostic strategies, and provides regional recurrence or distant metastasis. The tumor-infiltrating lymphocytes (TILs) are known as a critical diagnostic biomarker in antitumor immune response. We evaluated the association between CD4+ T-lymphocyte marker, some clinicopathological indices, and the impact of TILs on the stage and grade of OSCC. Materials and Methods: In this cross-sectional study, 37 OSCC specimens including 16 early and 21 advanced stages (categorized base-on recent clinical oncology references) and their related healthy surgical margin (as internal control group) were collected. Obtained histochemical data were analyzed by SPSS V.23 software. The expression of CD4+ marker in tumor microenvironment (TME) was compared by nonparametric Mann-Whitney and Kruskal-Wallis as well as Fisher's exact tests. P < 0.05 was remarked statistically significant. Results: The low-grade patients represented more CD4+ TIL that was statistically significant (P = 0.011). However, there was no statistically significant difference in CD4+ TIL between various stages (P = 0.404), tumor size, and lymph node involvement (P > 0.05). Moreover, there was no significant relation between TIL infiltration, age, and tumor localization (P > 0.05), however CD4+ expression in women was more than men (P = 0.008). The CD4+ T-lymphocyte infiltration in TME was more significant than healthy surgical margin (P < 0.001). There was a statistically significant difference between healthy surgical margin and different grades and stages of OSCCs that lower grades demonstrated more CD4+ TIL infiltration (P < 0.001). Conclusion: The CD4+ T-lymphocytes may play important role in differentiation and maturity of epithelial cell, tumorigenesis, and progression of OSCC.

The role of altered microRNA expression in premalignant and malignant head and neck lesions with epithelial origin.

Background and Aims: The premalignant lesions of the oral cavity carry a risk of transformation to malignancy. Hence, early diagnosis followed by timely intervention remarkably affects the prognosis of patients. During tumorigenesis, particular microRNAs (miRNAs) show altered expressions and because of their post transcriptionally regulatory role could provide favorable diagnostic, therapeutic, or prognostic values in head and neck cancers. Methods: In this review, we have demonstrated diagnostic, prognostic, and potential therapeutic roles of some miRNAs associated with oral premalignant and malignant lesions based on previous validate studies. Results: It is previously documented that dysregulation of miRNAs contributes to cancer development and progression. MiRNAs could be tumor suppressors that normally suppress cell proliferation, differentiation, and apoptosis or play as oncogenes that improved tumorigenesis process. Altered expression of miRNAs has also been reported in premalignant oral epithelial lesions such as leukoplakia, oral submucous fibrosis, oral lichen planus and some malignant carcinoma like oral squamous cell, verrucous, spindle cell, Merkel cell carcinoma and basal cell. Conclusion: Some of miRNAs could be new therapeutic candidates in miRNA-based target gene therapy. Although more investigations are required to identify the most favorable miRNA candidate, altered expression of some miRNAs could be used as biomarkers in premalignant lesions and oral cancers with high sensitivity and specificity.

Association between Tissue Expression of Toll-Like Receptor and Some Clinicopathological Indices in Oral Squamous Cell Carcinoma.

Background: The oral squamous cell carcinoma (OSCC) composes about 90% of all head and neck cancers. The toll-like receptor (TLR)+ immune cells have potential of invasion and malignancy transformation. The aim of this study was assessment of possible associations between clinicopathological indices and TLR2 and TLR9 gene expression in OSCC. Methods: Forty-two OSCC samples with related healthy margins including 25 early and 17 advanced stages were gathered. The samples were classified histologically from grade I to II. The expression of TLR2 and TLR2 was evaluated by Real-time PCR. The patient's disease-free survival (DFS) and overall survival (OS) were analyzed using SPSS V.23 software. Results: The expression of TLR2 and TLR9 genes in tumor tissues (especially in grade I and II) were higher than healthy surgical margin tissue (p< 0.001). TLR9 expression in grade II was statistically significant than grade I in tumor tissue (p< 0.001). TLR9 expression in advanced stage was statistically significant in compare to early stage (p= 0.012). In advanced stage both overall survival (p= 0.029) and disease-free survival (p= 0.012) were statistically lower than early stage. The follow-up time to recurrence in advanced stage was statistically lower than early stage (p= 0.007). Conclusion: Overexpression of TLRs 2, 9 play role in the pathogenesis and tumor development of OSCC and can be applied as biomarker in prognostic approaches.

Tumor tissue Helicobacter pylori and human papillomavirus infection in head and neck squamous cell carcinoma patients and association with clinicopathological indices: A cross-sectional medical survey.

Background: The associations between Helicobacter pylori and human papillomavirus (HPV) with head and neck squamous cell carcinomas (HNSCCs) are approved before. However, the association between demographic, clinicopathological, and histologic characteristics of HNSCC patients and molecular detection of HPV and H. pylori has not been enough investigated. Materials and Methods: In this cross-sectional study, 62 patients with HNSCC from January 2016 to February 2020 were entered the study. For H. pylori detection 16S ribosomal RNA and glmM genes and HPV detection, MY09 and MY11 genes were used. P < 0.05 is considered as significant level. Results: There were 34 patients with advanced-stage cancer (54.8%). Grade I patients (61.3%) had the highest frequency. There were 20 (32.25%) and 7 (11.29%) patients with positive H. pylori infection among tumor tissue and healthy tissue margins, respectively. Positive HPV infections were in 8 (12.90%) and 3 (4.83%) patients, respectively, in tumor tissue and healthy tissue margins (P = 0.01). There was a significant difference between histological grade and infection to HPV among HNSCC patients (P = 0.01), and most of the positive HPV cases had well-, moderate-, and poorly-differentiated tumors, respectively. Our study showed a significant increase in HPV infection in the advanced-stage group compared to the early-stage group (P = 0.05). Conclusion: Our study findings concluded a significant relationship between HPV infection in HNSCC patients with age, stage, and grade. In summary, our findings based on polymerase chain reaction analysis concluded remarkably a potential role of HPV infection and to some extent H. pylori infection into the contribution of HNSCC malignancies.

Evaluation of correlation between transcription factors and IL-17 in oral and cutaneous lichen planus lesions and Leukocytes.

BACKGROUND: Lichen planus (LP) is a chronic autoimmune disease with different clinical subtypes including cutaneous LP (CLP) and oral LP (OLP). We aimed to compare mRNA expression of RORγt and IL-17 in paraffin-embedded blocks of OLP and CLP lesions with normal oral mucosa (NOM), and also its correlation with hematologic parameters. MATERIALS & METHODS: This study included 89 paraffin-embedded blocks contain OLP (44 cases), CLP (45 cases) and NOM from the archive of Mashhad University of Medical Sciences, Mashhad, Iran. The expression of RORγt and IL-17 was evaluated by Real-time RT-PCR method. The result was compared to Leukocyte counts and the other hematological parameters of studied patients. RESULTS: The results of our study showed IL-17 and RORγt expression in OLP lesions were significantly higher than CLP and NOM groups (P = 0.001). Although we found high expression of RORγt and IL-17 in erosive OLP in compared to classic OLP lesion, but this increment was not significant for IL-17 (P = 0.26) and RORγt (P = 0.14). Further, Leukocyte and monocyte counts were substantially high in OLP group in compared to the CLP and NOM groups (P < 0.05). CONCLUSIONS: We concluded that increased expression of RORγt and IL-17 in LP lesions could play role in the pathogenesis of LP. As well, higher expression of RORγt and IL-17 in oral LP more than cutaneous LP might be associated with difference in clinical behavior of the two types of disease and role of these factors in premalignant behavior of OLP lesions.

Evaluation of the Relationship Between the Invasive Front of Oral Squamous Cell Carcinoma and Clinicopathological Parameters.

BACKGROUND & OBJECTIVE: The present study investigated the relationship between invasive front (IF) of tumors and clinicopathological parameters including stage, grade, nodal involvement, lymphocytic host response (LHR), recurrence, overall survival (OS), and disease-free survival (DFS). METHODS: A total of 87 oral squamous cell carcinoma (OSCC) biopsies were evaluated. Clinical stage, grading, nodal involvement, time of recurrence, OS, and DFS were assessed. The number of tumor budding cells in the IF was measured by two pathologists with an optic microscope. IF was graded to low risk (<5) and high risk (>5), according to the counting of tumor budding as a single cancer cell or cluster cells. Also, LHR was reported in the IF as mild, moderate, and severe. RESULTS: IF was reported in 43.7% of patients as a low-risk group and 49.4% as a high-risk group. LHR was also mild in 31%, moderate in 25.3%, and severe in 43.7% of the patients. Most of the patients were in stage IV (31%) and grade 1 (60.9%). The high risk IF group had a significant statistical relationship with stage (P=0.001), grade (P=0.039), five years OS (P=0.03), five years DSF (P=0.01), and lymph node involvement (P=0.007). The relation between LHR and stage of disease was significant (P=0.034). CONCLUSION: Considering the essential role of histopathological reports in the treatment plan of patients and the relationship between IF and clinical parameters, IF evaluation in routine histopathological examinations, especially in the early stages of OSCC, seems to be necessary.

Cytokeratins (CK7 and CK20) Genes Expression Association with Clinicopathological Indices in Oral Squamous Cell Carcinoma and Dysplastic Oral Epithelium.

BACKGROUND: High prevalence of oral squamous cell carcinoma (OSCC) demands the additional novel biological markers. Due to the established roles of cytokeratin in the prognosis of metastasis evaluation the relation of expression of both CK7 and CK20 in OSCC compared to the dysplastic oral epithelium biopsies with clinicopathological factors were investigated. METHODS: We examined the coordinate mRNA expression of cytokeratin 7 (CK7) and cytokeratin 20 (CK20) using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in 110 biopsies of oral squamous epithelium samples including 72 tumoral and 38 dysplastic biopsies. We also collected demographic and pathological data including tumor stage and grade from our patients. RESULTS: There was a significant difference in CK7 and CK20 gene expression between OSCC and dysplastic samples (p< 0.001). Further, their mean expression in OSCC samples was significantly higher compared to dysplastic samples. Relative mRNA levels of CK7 and CK20 showed that their mean expression in OSCC grade I was significantly lower than other grades (p< 0.01). The relationship between CK7 and CK20 mRNA expression and age or gender was not significant (p> 0.05). Samples in the advanced stage of disease had significantly higher CK7 and CK20 expression compared to early-stage samples of OSCC specimens (p= 0.001). CONCLUSION: We found an increase in CK7 and CK20 mRNA levels in grade III OSCC samples compared to other grades. This finding suggests a potential role for CK7 and CK20 in oral mucosal carcinogenesis and OSCC prognosis.

LAMP3 (CD208) Expression in Squamous Cell Carcinoma and Epithelial Dysplasia of the Oral Cavity and Clinicopathological Characteristics of Unfavorable Prognosis.

BACKGROUND: This study aimed to evaluate LAMP3 (CD208) gene expression in oral squamous cell carcinoma (OSCC) and dysplastic oral epithelium by quantitative real-time polymerase chain reaction (qPCR) and compare LAMP3 expression in different disease grades and stages. METHODS: In this study, 60 OSCC and dysplastic oral epithelium samples were obtained from the Mashhad University of Medical Sciences together with their demographic and clinicopathological documents. LAMP3 expression was measured by qPCR. RESULTS: LAMP3 expression was significantly greater in OSCC than in dysplasia samples (P=0.001), in grade III OSCC than in grades I and II, and also greater in advanced than in early OSCC disease stage (P=0.001). CONCLUSION: The significantly greater LAMP3 expression in OSCC than in dysplastic epithelium indicates a role for LAMP3 in carcinogenesis in oral mucosa. Our results suggest LAMP3 may be useful as an anticancer target and/or to predict disease pathogenesis in OSCC patient's cells.

New diagnostic molecular markers and biomarkers in odontogenic tumors.

Odontogenic tumors comprised of complex heterogeneous lesions that diverse from harmatomas to malignant tumors with different behavior and histology. The etiology of odontogenic tumors is not exactly determined and pathologists deal with challenges in diagnosis of odontogenic tumors because they are rare and obtained experiences are difficult to evaluate. In this study, we describe immunohistochemical and molecular markers in diagnosis of odontogenic tumors besides advanced diagnostic technique. Immunohistochemical features of odontogenic tumors beside the clinical features and radiological finding can help us to determine the correct diagnosis. Although these markers are neither specific nor sensitive enough, but analysis of gene expression provides definitive confirmation of diagnosis. In addition, "-omics" technology detected specific molecular alternation associated with etiology such as genomics, epigenomics, transcriptomics, proteomics and metabolomics. The post transcriptional events such as DNA methylation and chromatin remodeling by histone modification affect the changes in epigenome. Furthermore, non-coding RNAs like micro-RNAs, long noncoding RNA (lncRNA) and small non-coding RNA (snoRNA) play regulatory role and impact odontogenesis. Molecular marker propose their potential role in etiopathogenesis of odontogenic tumors and suitable candidate in diagnostic, prognostic and therapeutic approaches in addition to patient management. For future evaluations, organoid represents in vitro tumor model-study for tumor behavior, metastasis and invasion, drug screening, immunotherapy, clinical trial, hallmarks association with prognosis and evolution of personalized anti-cancer therapy. Moreover, organoid biobank help us to check genetic profile. We think more investigation and studies are needed to gain these knowledges that can shift therapeutic approaches to target therapy.

Potential therapeutic approaches of microRNAs for COVID-19: Challenges and opportunities.

The coronavirus disease 2019 (COVID-19) emerges as current outbreak cause by Novel Severe Acute Respiratory Syndrome Corona Virus-2 (SARS-CoV-2). This infection affects respiratory system and provides uncontrolled systemic inflammatory response as cytokine storm. The main concern about SARS-CoV-2 pandemic is high viral pathogenicity with no specific drugs. MicroRNAs (miRs) as small non-coding RNAs (21-25 â€‹nt) regulate gene expression. The SARS-CoV-2 encoded-miRs affect human genes that involved in transcription, translation, apoptosis, immune response and inflammation. Also, they alter self-gene regulation and hijacked host miRs that provide protective environment to maintain its latency. On the other hand, Host miRs play critical role in viral gene expression to restrict infection. Over expression/inhibition of miRs might result in cell cycle irregularity, impaired immune response or cancer. In this manner, exact role of each miR should be specified. Mimic encoded-miRs like antagomirs showed successful result in phases of clinical trial prevent from negative effects of viral encoded-miRs. Products of mimic miRs are inexpensive corresponds to synthesis of primer; they are short and nanoscale in size. Although SARS-CoV-2 genome is undergoing evaluation, detection of exact molecular pathogenesis open up opportunities to for vaccine development. Salivaomics can evaluate SARS-CoV-2 genome, transcriptome, proteome and biomarkers like miRs in oral related and cancer disease. In this review, we studied the challenge and opportunities of miRs in therapeutic approach for SARS-CoV-2 infection, then overviewed the role of miRs in saliva droplet during SARS-CoV-2 infection and related cancer.