Oral Squamous Carcinoma Research Articles

Can Radiological Tumour Thickness Predict Pathological Prognostic Factors in Clinicoradiologically Node-Negative Oral Squamous Carcinoma? A Prospective Study.

This research investigates potential connections between radiological tumour thickness determined by CT scans and various pathological prognostic factors. These factors include pathological tumour thickness (pTT), pathological depth of invasion (DOI), and positive cervical nodal metastasis. This analysis focuses on cases of clinicoradiologically node-negative squamous cell carcinoma of the buccal mucosa. Sixty-one previously untreated clinicoradiologically node-negative squamous cell carcinoma of buccal mucosa were included in the study. The radiological tumour thickness in the preoperative CT scans is correlated with other prognostic factors like pathological tumour thickness, DOI and presence or absence of neck node. Sixty-one patients were included in the study with a median age of 54 years (Range 27-84). Forty-two patients (68.9%) were male, and 19 were females (31.1%). There was no statistically significant difference in mean values of rTT among patients with positive or negative post-operative nodal metastases. However, a significant correlation could be established with rTT to other potential prognostic factors. Tumor thickness in preoperative CT scans can be used to predict post-operative prognostic factors in oral squamous cell carcinoma.

Serological profile of IGG antibodies to viral capsid antigen of epstein-barr virus in oral pre-cancer and cancer patients

Cancer of the oral cavity is a major and increasing public health problem. Oral cancer is the sixth most common cancer reported globally with an annual incidence of over 300,000 cases, of which 62% arise in developing countries. In India, because of cultural, ethnic, and geographic factors and the popularity of addictive habits, the frequency of oral cancer is high. Despite the various etiological and predisposing factors associated with oral pre-cancer and cancer, the role of co-carcinogenic effects of viruses like HSV has been documented. With this background and considering the high prevalence of oral cancer in India an attempt has been made to study the association of EBV with oral cancer using serological methods.1. To study the IgG antibodies to Viral Capsid Antigen (VCA) of Epstein-Barr virus in oral pre-cancer and cancer by a laboratory standardized Indirect Immuno Fluorescent (IIF) test. 2. To compare and corelate the results of IIF test with a commercially available ELISA for VCA IgG antibodies to EBV. 3. To assess the severity of disease progression in oral pre-cancer and cancer using the serological profile.In the present study, 84 subjects were assessed for serum IgG antibodies to VCA of EBV using the IIF technique and later the results were compared with an ELISA kit. Group I were controls, Group II to Group V were oral squamous cell carcinoma subjects according to the clinical staging. Group VI comprised of oral pre-cancer subjects. Age range was between 40-70 years, and the mean age was 50.31 +/- 8.23 years. All the 42 control sera were positive for IgG antibodies to VCA of EBV but with a significantly low geometric mean titre(GMT) of 8.561.The GMTs of oral squamous carcinoma and pre-cancer sera was positive but progressively increased when compared to control sera and were highly significant (P<0.001).As there was a significant increase in the GMTs in oral squamous cell carcinoma subjects and pre-cancer subjects, serology of EBV may be interpreted to be useful in assessing the severity of the disease and the techniques of IIF and ELISA may be used as first line screeners when large number of subjects need to be assessed.

Comparative Transcriptome Analysis Identifies Desmoglein-3 as a Potential Oncogene in Oral Cancer Cells.

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced, and 599 candidate genes exhibited differential expression between DSG3-overexpressing and matched control lines, with 12 genes highly significantly differentially expressed, including 9 upregulated and 3 downregulated. Genes with known implications in cancer, such as MMP-13, KRT84, OLFM4, GJA1, AMOT and ADAMTS1, were strongly linked to DSG3 overexpression. Gene ontology analysis indicated that the DSG3-associated candidate gene products participate in crucial cellular processes such as junction assembly, focal adhesion, extracellular matrix formation, intermediate filament organisation and keratinocyte differentiation. Validation of RNA-Seq was performed through RT-qPCR, Western blotting and immunofluorescence analyses. Furthermore, using transmission electron microscopy, we meticulously examined desmosome morphology and revealed a slightly immature desmosome structure in DSG3-overexpressing cells compared to controls. No changes in desmosome frequency and diameter were observed between the two conditions. This study underscores intricate and multifaceted alterations associated with DSG3 in oral squamous carcinoma cells, implying a potential oncogenic role of this gene in biological processes that enable cell communication, motility and survival.

New Megastigmane and Polyphenolic Components of Henna Leaves and Their Tumor-Specific Cytotoxicity on Human Oral Squamous Carcinoma Cell Lines.

Polyphenols have a variety of phenolic hydroxyl and carbonyl functionalities that enable them to scavenge many oxidants, thereby preserving the human redox balance and preventing a number of oxidative stress-related chronic degenerative diseases. In our ongoing investigation of polyphenol-rich plants in search of novel molecules, we resumed the investigation of Lawsonia inermis L. (Lythraceae) or henna, a popular ancient plant with aesthetic and therapeutic benefits. The leaves' 70% aq acetone extract was fractionated on a Diaion HP-20 column with different ratios of H2O/an organic solvent. Multistep gel chromatographic fractionation and HPLC purification of the Diaion 75% aq MeOH and MeOH fractions led to a new compound (1) along with tannin-related metabolites, benzoic acid (2), benzyl 6'-O-galloyl-β-D-glucopyranoside (3), and ellagic acid (4), which are first isolated from henna. Repeating the procedures on the Diaion 50% aq MeOH eluate led to the first-time isolation of two O-glucosidic ellagitannins, heterophylliin A (5), and gemin D (6), in addition to four known C-glycosidic ellagitannins, lythracin D (7), pedunculagin (8), flosin B (9), and lagerstroemin (10). The compound structures were determined through intensive spectroscopic investigations, including HRESIMS, 1D (1H and 13C) and 2D (1H-1H COSY, HSQC, HMBC, and NOESY) NMR, UV, [α]D, and CD experiments. The new structure of 1 was determined to be a megastigmane glucoside gallate; its biosynthesis from gallic acid and a β-ionone, a degradative product of the common metabolite β-carotin, was highlighted. Cytotoxicity investigations of the abundant ellagitannins revealed that lythracin D2 (7) and pedunculagin (8) are obviously more cytotoxic (tumor specificity = 2.3 and 2.8, respectively) toward oral squamous cell carcinoma cell lines (HSC-2, HSC-4, and Ca9-22) than normal human oral cells (HGF, HPC, and HPLF). In summary, Lawsonia inermis is a rich source of anti-oral cancer ellagitannins. Also, the several discovered polyphenolics highlighted here emphasize the numerous biological benefits of henna and encourage further clinical studies to profit from their antioxidant properties against oxidative stress-related disorders.

Fabrication of a Three-Dimensional Spheroid Culture System for Oral Squamous Cell Carcinomas Using a Microfabricated Device.

Cancer stem cells (CSCs) are considered to be responsible for recurrence, metastasis, and resistance to treatment in many types of cancers; therefore, new treatment strategies targeting CSCs are attracting attention. In this study, we fabricated a polyethylene glycol-tagged microwell device that enabled spheroid formation from human oral squamous carcinoma cells. HSC-3 and Ca9-22 cells cultured in the microwell device aggregated and generated a single spheroid per well within 24-48 h. The circular shape and smooth surface of spheroids were maintained for up to five days, and most cells comprising the spheroids were Calcein AM-positive viable cells. Interestingly, the mRNA expression of CSC markers (Cd44, Oct4, Nanog, and Sox2) were significantly higher in the spheroids than in the monolayer cultures. CSC marker-positive cells were observed throughout the spheroids. Moreover, resistance to cisplatin was enhanced in spheroid-cultured cells compared to that in the monolayer-cultured cells. Furthermore, some CSC marker genes were upregulated in HSC-3 and Ca9-22 cells that were outgrown from spheroids. In xenograft model, the tumor growth in the spheroid implantation group was comparable to that in the monolayer culture group. These results suggest that our spheroid culture system may be a high-throughput tool for producing uniform CSCs in large numbers from oral cancer cells.

PRF Lysates Enhance the Proliferation and Migration of Oral Squamous Carcinoma Cell Lines.

Platelet-rich fibrin (PRF) is an autologous fibrin-rich matrix where activated platelets and leucocytes accumulate. PRF has a wide spectrum of clinical indications with the overall aim of supporting tissue regeneration which in dentistry includes the healing of healthy oral mucosa with epithelial cells. In oral squamous cell carcinoma lesions, however, epithelial cells undergo malignant transformation, indicated by their unrestricted proliferation and migration potential, which should not be further enhanced by a wound-healing formula. Yet, little is known about how oral squamous cell carcinomas respond to PRF lysates. The aim of the present study was, therefore, to test the capacity of PRF lysates to change the transcriptome of HSC2 oral squamous carcinoma cells and perform bioassays to support the findings. Based on the RNAseq analysis, PRF lysates caused an increase in the genes functionally linked to cell replication and migration. In support of this screening approach, PRF lysates enhanced the proliferation of HSC2 oral squamous carcinoma cells, as indicated by 3[H]-thymidine incorporation, cell counting, and the expression of proliferation-related genes. Moreover, PRF lysates sped up cell migration in a scratch assay requiring actin polymerization. Taken together, our data showing that PRF lysates are mitogenic and stimulate motility of oral squamous carcinoma cell lines could be an indication that treatment with PRF in cases of oral carcinoma should be carefully considered.

Clinical Experience with Autofluorescence Guided Oral Squamous Cell Carcinoma Surgery.

In our study, the effect of the use of autofluorescence (Visually Enhanced Lesion Scope-VELscope) on increasing the success rate of surgical treatment in oral squamous carcinoma (OSCC) was investigated. Our hypothesis was tested on a group of 122 patients suffering from OSCC, randomized into a study and a control group enrolled in our study after meeting the inclusion criteria. The preoperative checkup via VELscope, accompanied by the marking of the range of a loss of fluorescence in the study group, was performed before the surgery. We developed a unique mucosal tattoo marking technique for this purpose. The histopathological results after surgical treatment, i.e., the margin status, were then compared. In the study group, we achieved pathological free margin (pFM) in 55 patients, pathological close margin (pCM) in 6 cases, and we encountered no cases of pathological positive margin (pPM) in the mucosal layer. In comparison, the control group results revealed pPM in 7 cases, pCM in 14 cases, and pFM in 40 of all cases in the mucosal layer. This study demonstrated that preoperative autofluorescence assessment of the mucosal surroundings of OSCC increased the ability to achieve pFM resection 4.8 times in terms of lateral margins.

Ganoderma lucidum polysaccharides associated with 5-Fluorouracil impair OSCC tumorigenesis in vitro

BackgroundsGanoderma lucidum polysaccharides have been shown several anti-cancer, anti-inflammatory, and immunomodulatory properties among studies with different tumor models and its use with advanced and conventional combination therapies is a world trend. The administration of 5-Fluorouracil is already used as chemotherapy for many tumors and works on tumor remission; however, its adverse effects are still severe, impoverishing treatment and quality of life for patients. Cancer stem cells represent a subpopulation of cells with defense mechanisms against chemotherapy agents and are the main cause of relapses and metastases in cancer treatments. Also, the Epithelial-Mesenchymal Transition program increases properties related to tumor malignancy and mortality. In this scenario, the aim of the study is to evaluate the effects of Ganoderma lucidum polysaccharides in combination with 5-Fluorouracil on the subpopulation of cancer stem cells present in the human oral squamous carcinoma cell line SCC-9. MethodsSCC-9 cells were treated in vitro for 72 h with different 5-Fluorouracil low doses, associated or not with Ganoderma lucidum polysaccharides. Cells maintained with culture media or cisplatin were used as control. All the cells were evaluated for cytotoxicity, cancer stem cells, and Epithelial-Mesenchymal Transition properties. ResultsThe associated treatment avoided proliferation, delayed migration, slightly modified morphology of cells, increased apoptosis, decreased colony and blocked spheres formation, and downregulated cancer stem cells, Epithelial-Mesenchymal Transition, and ABC drug transporters expression. In addition, Ganoderma lucidum polysaccharides + 5-Fluorouracil changed the treated cells into a non-cancer stem cell phenotype, a characteristically not resistant and less proliferative population. The 5-Fluorouracil treatment alone showed remarkable modification in cellular morphology, apoptosis, and absence of holoclones; however, it upregulated the molecular expression of cancer stem cells' hallmarks. ConclusionsThese findings demonstrate that combining Ganoderma lucidum polysaccharides with a low dose of 5-Fluorouracil is effective against oral squamous cell carcinoma in vitro by enhancing the cells' sensitivity to drugs and reducing the characteristics associated with cancer stem cells (CSCs). This suggests the possibility of reducing conventional chemotherapy doses and improving oral squamous cell carcinoma treatment. It also highlights the potential for this combination to be used as an adjunct in Complementary Alternative Medicine (CAM).

Natural product piperlongumine inhibits proliferation of oral squamous carcinoma cells by inducing ferroptosis and inhibiting intracellular antioxidant capacity.

As a new form of cell death, ferroptosis has been shown to have inhibitory effects on a variety of tumor cells except oral squamous cell carcinoma (OSCC). There were few investigations on the effects and molecular mechanisms of piperlongumine (PL, a ferroptosis inducer) and CB-839 (a GLS1 inhibitor which promotes ferroptosis) on OSCC cells. This article assesses the anticancer effect and mechanism of PL as well as combined with CB-839. OSCC cells were treated with specified concentration of PL alone or with ferroptosis inhibitor Ferrostatin-1 (Fer-1) and antioxidant N-Acetylcysteine (NAC) to assess their effects on biological characteristics such as cell proliferation, cell death and intracellular ferroptosis related pathways. Also, cells were treated with PL combined with CB-839 to evaluate the synergistic effect of CB-839 on PL's anticancer effects. The results showed that the proliferation rate of PL-treated OSCC cells were decreased in a dose- and time-dependent manner. PL can induce OSCC cells apoptosis. Lipid peroxidation (LPO) and intracellular reactive oxygen species (ROS) were accumulated after PL treatment. We found some protein changes significantly such as the expression of DMT1 increased, and the expression of FTH1, SLC7A11 and GPX4 decreased. In addition, the anti-proliferation effect of PL can be reversed by Fer-1 and NAC and the level of LPO and ROS was decreased accordingly. Importantly, we found that PL and CB-839 in combination could decrease the cell viability and the LPO level synergistically, accompanied by a large consumption of glutathione (GSH). These evidences prove that PL can induce ferroptosis of OSCC cells, which can be enhanced by CB-839. Our study suggested that the nature product PL can induce the ferroptotic death of OSCC cells, which is further enhanced when combined with CB-839. The synergistic anticancer effect of these two may prove new strategy for OSCC treatment.

Abstract PO-031: Integrated bioinformatics and functional spatial analysis platform for the discovery of predictive biomarkers in oral cancer

Abstract Background: Oral cavity squamous cell carcinoma (OCSCC) has an unpredictable response to immunotherapy, and standard biomarkers evaluating the tumor immune microenvironment (TIME) lack widespread translational applicability. Defining the TIME is critical to understanding mechanisms and allows the development of reproducible and accurate prognostic and predictive biomarkers based on immune profiles; however, to date, this remains elusive for clinical use. To further study and deconstruct the TIME within OCSCC, we integrated our previously developed computational spatial analysis (CSA) with in-silico deconvolution analyses to comprehensively study the TIME in OCSCC. Methods: We leveraged access to two independent OCSCC cohorts (UM and TCGA) with paired bulk RNASeq, WES, and CSA-immune score data. TMB, neoantigen load, and deconvolution analysis (CIBERSORTx) were performed to generate in silico immune profiles. A previously described CSA evaluates the density and distribution of tumor-infiltrating lymphocytes (TILs) relative to cancer cells to generate immune scores (GCross function [GFx]). GFx scores were then correlated with WES and RNASeq-derived cell-type deconvolution results, immune cell activation expression signatures, and gene expression-based radiosensitivity index scores. Pearson's correlation coefficient and associated p-value were calculated for each comparison. Cox proportional hazards multivariate regression was performed using the entire cohort for survival and recurrence analysis. Results: Here, we better characterize the immunologic tumor environment by objectively measuring the TIL density in spatial relationships with cancer cells. We found that immunoreactive tumors with high GFx scores are highly prognostic for overall survival, with the lowest tiers associated with poor survival. Combining the spatial relationship with immunogenomics (WES and RNASeq immune profiles) increased the prognostic capability of the integrated bioinformatics analysis model. In addition, we further revealed that lymphocyte-infiltrated regions, specifically CD8+ T-cells, are associated with TMB, neoantigen load, and improved overall survival. Conclusion: This study provides an integrated dataset from which to generate future insights and possible immunomodulatory therapies to counteract the poor immunogenicity of OCSCC. We show that integrating computational spatial analysis and in-silico deconvolution analysis is generalizable to multiple patient populations with different demographic compositions and pathology images collected from distinctive digitization methods. Our machine-learning approaches provide clinically actionable predictions that confirm the relevance of these biomarkers with immune cell infiltration and suggest the role of digital pathology profiling in predicting response to immunotherapy which could inform treatments for oral squamous carcinoma patients. Citation Format: Michael M. Allevato, Lisa Chionis, Joshua Smith, Santhoshi Krishnan, Leanne Henry, Tingting Qin, Janice Farlow, Laura Rozek, Jonathan Mchugh, Arvind Rao, Maureen Sartor, Steven B. Chinn. Integrated bioinformatics and functional spatial analysis platform for the discovery of predictive biomarkers in oral cancer [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-031.

Unveiling the anti-cancer mechanisms of calotropin: Insights into cell growth inhibition, cell cycle arrest, and metabolic regulation in human oral squamous carcinoma cells (HSC-3)

BackgroundCalotropin, a cardiac glycoside obtained from the plant Calotropis gigantea, has demonstrated promising potential as an anti-tumorigenesis compound. ObjectiveThe main objective of this study was to investigate the potential anti-cancer properties of calotropin against HSC-3 oral squamous cancer cells and to elucidate the underlying mechanisms involved in its action. Material and methodCalotropin were treated in HSC-3 to evaluate cell viability by MTT assay. Flow cytometry analysis divulged that calotropin G0/G1 phase cell cycle arrest and apoptosis in HSC-3 cells. Calotropin displayed inhibitory properties against aerobic glycolysis, a metabolic alteration using glucose uptaken, lactose production and LDHA activity assays. Furthermore, migration and invasion assays help that calotropin has ability to reduce the migratory and invasive of HSC-3 cells, using transwell and Matrigel assay. Validation of mRNA expression through RT-PCR. Molecular docking was implemented to validate the binding association of calotropin with apoptosis and metastatic regulating targets. ResultThe results exemplify that increasing doses of calotropin effectively hold back the HSC-3 cell progression. Migration and invasion assays help that calotropin has ability to reduce the migratory and invasive of HSC-3 cells, indicating its potential to inhibit cancer metastasis. These results imply that calotropin may influence genes linked to metastasis and apoptosis in order to achieve its beneficial effects on cancer. Docking results provided further support, showing a high binding energy between calotropin and metastasis-mediated pathways. ConclusionOverall, our findings shed an experimental evidence on how calotropin inhibits the HSC-3 oral squamous cancer cell growth, highlighting the drug's potential as a treatment for oral cancer. Further, investigation on in-vivo experiment is warranted to explore its potential mechanism of action and to develop a novel drug towards clinical trial.

Enamel Matrix Derivative Suppresses Chemokine Expression in Oral Epithelial Cells.

Epithelial cells in periodontitis patients increasingly express chemokines, suggesting their active involvement in the inflammatory process. Enamel matrix derivative (EMD) is an extract of porcine fetal tooth germs clinically applied to support the regrowth of periodontal tissues. Periodontal regeneration might benefit from the potential anti-inflammatory activity of EMD for epithelial cells. Our aim was, therefore, to set up a bioassay where chemokine expression is initiated in the HSC2 oral squamous carcinoma cell line and then test EMD for its capacity to lower the inflammatory response. To establish the bioassay, HSC2 cells being exposed to TNFα and LPS from E. coli (Escherichia coli) or P. gingivalis (Porphyromonas gingivalis) were subjected to RNAseq. Here, TNFα but not LPS caused a robust increase of chemokines, including CXCL1, CXCL2, CXCL8, CCL5, and CCL20 in HSC2 cells. Polymerase chain reaction confirmed the increased expression of the respective chemokines in cells exposed to TNFα and IL-1β. Under these conditions, EMD reduced the expression of all chemokines at the transcriptional level and CXCL8 by immunoassay. The TGF-β receptor type I kinase-inhibitor SB431542 reversed the anti-inflammatory activity. Moreover, EMD-activated TGF-β-canonical signaling was visualized by phosphorylation of smad3 and nuclear translocation of smad2/3 in HSC2 cells and blocked by SB431542. This observation was confirmed with primary oral epithelial cells where EMD significantly lowered the SB431542-dependent expression of CXCL8. In summary, our findings suggest that TGF-β signaling mediates the effects of EMD to lower the forced expression of chemokines in oral epithelial cells.

A Retrospective 2-D and 3-D Radiographic analysis of Osseous Changes in Oral Squamous Carcinoma

Aim: To evaluate the 2D and 3D radiographic features of oral squamous cell carcinoma involving jaws for understanding the pattern of bone involvement, for an improved treatment plan. Materials and Methods: Thirty histologically diagnosed patients of oral squamous cell carcinoma were included in the study and the detailed analysis of Orthopantogram (OPG), Computed Tomography (CT)/Cone Beam Computed Tomography (CBCT) / Magnetic Resonance Imaging (MRI) was undertaken. Lesions borders, surrounding and internal structures were evaluated. Results: 2D images aid in identifying the location of the lesion while 3D images help in defining the extension of the lesion. 2D images result in overdiagnosis of extent while 3D images provide an accurate diagnosis of the structures involved. Conclusions: Early screening of the lesion can be done by OPG while CBCT/CT/MRI should be considered as a confirmatory tool to see the extent of disease and for guidance in surgical planning.

Redox-Sensitive Delivery of Doxorubicin from Nanoparticles of Poly(ethylene glycol)-Chitosan Copolymer for Treatment of Drug-Resistant Oral Cancer Cells.

Reactive oxygen species (ROS)-sensitive polymer nanoparticles were synthesized for tumor targeting of an anticancer drug, doxorubicin (DOX). For this purpose, chitosan-methoxy poly(ethylene glycol) (mPEG) (ChitoPEG)-graft copolymer was synthesized and then DOX was conjugated to the backbone of chitosan using a thioketal linker. Subsequently, the chemical structure of the DOX-conjugated ChitoPEG copolymer (ChitoPEGthDOX) was confirmed via 1H nuclear magnetic resonance (NMR) spectra. Nanoparticles of the ChitoPEGthDOX conjugates have spherical shapes and a size of approximately 100 nm. Transmission electron microscopy (TEM) has shown that ChitoPEGthDOX nanoparticles disintegrate in the presence of hydrogen peroxide and the particle size distribution also changes from a monomodal/narrow distribution pattern to a multi-modal/wide distribution pattern. Furthermore, DOX is released faster in the presence of hydrogen peroxide. These results indicated that ChitoPEGthDOX nanoparticles have ROS sensitivity. The anticancer activity of the nanoparticles was evaluated using AT84 oral squamous carcinoma cells. Moreover, DOX-resistant AT84 cells were prepared in vitro. DOX and its nanoparticles showed dose-dependent cytotoxicity in both DOX-sensitive and DOX-resistant AT84 cells in vitro. However, DOX itself showed reduced cytotoxicity against DOX-resistant AT84 cells, while the nanoparticles showed almost similar cytotoxicity to DOX-sensitive and DOX-resistant AT84 cells. This result may be due to the inhibition of intracellular delivery of free DOX, while nanoparticles were efficiently internalized in DOX-resistant cells. The in vivo study of a DOX-resistant AT84 cell-bearing tumor xenograft model showed that nanoparticles have higher antitumor efficacy than those found in free DOX treatment. These results may be related to the efficient accumulation of nanoparticles in the tumor tissue, i.e., the fluorescence intensity in the tumor tissue was stronger than that of any other organs. Our findings suggest that ChitoPEGthDOX nanoparticles may be a promising candidate for ROS-sensitive anticancer delivery against DOX-resistant oral cancer cells.

A Method by which circUBR1 Controls the Proliferation, Migration, and Invasion of HSC3 Cells

This work investigates the effects of circUBR1 and the underlying mechanisms on the growth, migration and penetration of HSC3 cells derived from oral squamous carcinoma. The expression levels of circUBR1, miR-216a-3p and HSC3 were assessed in tissues from oral squamous cell carcinoma (OSCC) and adjacent tissues.
 Based on the transfection targets, the cells were divided into four groups: si-NC, si-circUBR1, miR-NC, miR-216a-3p, si-circUBR1+anti-miR-NC and si-circUBR1+anti-miR-216a-3p. The cell viability, scratch healing rate, MMP-2 and MMP-9 expression, number of cell clones formed, and number of invasion cells were significantly lower in the miR-216a-3p group compared to the miR-NC group (p<0.05).
 Moreover, when comparing the si-circUBR1+anti-miR-NC group to the si-circUBR1+anti-miR-216a-3p group, the cell viability, scratch healing rate, MMP-2 and MMP-9 expression, number of cell clones formed, and number of invasion cells were significantly higher in the si-circUBR1+anti-miR-216a-3p group (p<0.05). This suggests that disrupting circUBR1 expression leads to up-regulation of miR-216a-3p, resulting in reduced proliferation, clone formation, migration, and invasion of OSCC cells.

Association of Age and Frailty With 30-Day Outcomes Among Patients Undergoing Oral Cavity Cancer Surgery.

To evaluate the impact of age and frailty on 30-day outcomes following surgery for oral squamous cavity carcinoma (OSCC). Retrospective cross-sectional analysis. American College of Surgeons' National Quality Improvement Program (NSQIP) database. Patients who underwent OSCC resection were queried via NSQIP (2015-2020). Cases were stratified by age (18-65, 65-75, and older than 75) as well as by modified frailty index scores (mFI 0, mFI 1, and mFI 2+) for comparative analyses. Univariate and multivariable analyses were conducted to examine demographics, perioperative outcomes, and 30-day postoperative adverse events. A total of 3238 patients who underwent OSCC surgery were identified and categorized as nongeriatric ("NGA," age 18-65), younger geriatric ("YGA," age 65-75), and older geriatric ("OGA," age >75) adults. Compared to NGA, geriatric patients had higher the American Society of Anesthesiologists classification, higher modified frailty index scores, and more comorbidities such as hypertension, congestive heart failure, chronic obstructive disease, and diabetes (p < .001). YGAs and OGAs were also less likely to undergo neck dissection (p < .001), composite resection (p = .006), and free flap reconstruction compared to NGAs (p < .001). When controlling for confounders, age was not independently associated with an increased risk of poor outcomes. On the other hand, frailty was found to be independently associated with a higher risk of adverse events (odds ratio: 1.40 [1.15-1.70], p < .001 for mFI 1, odds ratio: 1.45 [1.04-2.02], p = .027 for mFI 2+). A higher mFI score, not older age, is associated with an increased risk of 30-day complications following OSCC surgery.

UPLC–ESI–QTOF–MS profiling, antioxidant, antidiabetic, antibacterial, anti-inflammatory, antiproliferative activities and in silico molecular docking analysis of Barleria strigosa

BackgroundThis study investigated the in vitro antidiabetic, antioxidant, antibacterial, anti-inflammatory and antiproliferative effects of B. strigosa hydrophilic (BSTR) and lipophilic (LSB) leaves extracts. The phytochemical profile was also performed using UHPLC–ESI–QTOF–MS.ResultsThe results indicated that BSTR and LSB showed excellent antioxidant properties in the DPPH scavenging, ABTS scavenging, FRAP and MCA assays. The extracts also demonstrated α-glucosidase (81.56–157.56 µg/mL) and α-amylase (204.44 µg/mL) inhibitory activities. In addition, the extracts showed significant cytotoxic and antiproliferative effects against oral squamous carcinoma (CLS-354/WT) cancer cells. Furthermore, the extracts showed excellent antibacterial activity against Listeria monocytogenes, Vibrio parahaemolyticus, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Both extracts exhibited a significant reduction in nitric oxide secretion against activated macrophage cells. The UHPLC–MS analysis revealed that B. strigosa is rich in terpenoids, iridoid glycosides, flavonoids, and phenolic compounds. The plethora of these compounds may be responsible for the observed activities. In addition, the bioactive compounds identified by UHPLC–ESI–QTOF–MS were analyzed using silico molecular docking studies to determine the binding affinity with α-amylase and α-glucosidase.ConclusionsThese results suggest that B. strigosa is an excellent pharmacological active plant and it provides the basis for further studies on the exploration of its potentials in oxidative stress induced disorders.Graphical

Identification of copy number alternation profiles in metastatic oral squamous carcinoma patients by using microarray-based comparative genomic hybridization: A study on Turkish patients

ObjectiveOral squamous cell carcinoma (OSCC) is a severe form of cancer affecting different anatomic sites of the oral cavity. OSCC ranks as the sixth most common cancer type with an increasing prevalence globally. However, the mechanisms of OSCC process at later stages are not well understood. In this study, we aimed to determine genetic alternations in metastatic OSCC patients to identify genomic changes occurred at metastatic phase of the disease. Material and MethodsThe Illumina CytoSNP-12 Array was used to determine copy number variations in OSCC cancer genome. Hybridization procedures were performed according to the manufacturer procedures (Illumina). Arrays were scanned on iScan System (Illumina). Data were analyzed using Illumina Genotyping module of Genome Studio software (version 1.2, Illumina). Multiple CNV algorithms and copy number alternations were accessed by Genome Studio. CNVs in whole genome were investigated by using a chromosomal heat map. ResultsWe reported that gains in 8q21.11-ter, 9p21.3, 13q14.11-ter, 13q13.3-ter and losses in 5q14.3-ter, 5q35 and 17p13.3–12 were associated with the development of OSCC. In addition, we also detected that deletion in 2q33.2-ter and 2q35–37.3 regions were also associated with OSCC metastasis process. ConclusionsOur results were also showed that gains in 11q13.3-q13.4 and 2q13.2 chromosomal regions could promote the metastatic OSCC process. We believe that results of the study will help to find new biomarkers for diagnosis at later stage of OSCC.

Synergistic effect of the combination of Chrysanthemum cinerariifolium (Trev.) and doxorubicin in inhibiting PI3K and Cyclin D in oral squamous cell carcinoma: in vitro study

Background: The most common type of lips and oral cavity cancer is oral squamous cell carcinoma (OSCC). Doxorubicin (DX) is commonly used as a chemotherapy drug, but its use is limited due to risk factors and drug resistance. Chrysanthemum cinerariifolium (Trev.) (CC) has potential as an anticancer agent. Combining the plant extract and chemotherapy drug might prevent OSCC proliferation by inhibiting PI3K and cyclin D protein. Therefore, the present study aimed to determine the synergistic effect of the combination of C. cinerariifolium (Trev.) and doxorubicin in inhibiting PI3K and Cyclin D protein. Methods: Human oral squamous carcinoma cell lines SCC-9 were used in this study. A cytotoxicity assay was performed to obtain the IC50 value of CC ethanol extract and DX on the SCC-9 cell line. Synergism evaluation of the combination CC and DX was analyzed using CompuSyn software. ELISA and the immunofluorescent assay were performed to determine the level of PI3K and cyclin D in the SCC-9 cell line after being treated with IC50 value of CC, IC50 value of DX and three combinations of CC and DX [7/8 IC50 CC + 1/8 IC50 DX (dose 1), 6/8 IC50 CC + 2/8 IC50 DX (dose 2), and 4/8 IC50 CC + 4/8 IC50 DX (dose 3). Results: CC stem ethanol extract and DX inhibited the proliferation of SCC-9 cell lines with the IC50 value of 133.4 µg/mL and 288.3 nM, respectively. The combination of CC and DX at dose 2 (6/8 IC50 CC + 2/8 IC50 DX) exhibited a high decrease in PI3K and cyclin D expression. Conclusions: The combination of C. cinerariifolium and doxorubicin synergistically declined OSCC proliferation by inhibiting PI3K and cyclin D expression.

Integrin β1 regulates the perineural invasion and radioresistance of oral squamous carcinoma cells by modulating cancer cell stemness

Perineural invasion and radioresistance are the main determinants of treatment outcomes in oral squamous cell carcinoma (OSCC), but the exact mechanism is still unknown. We conducted an in vitro experiment to evaluate the role of integrin β1 (ITGB1) in the perineural invasion, radioresistance, and tumor aggressiveness of OSCC. Two OSCC cell lines (SCC25, SCC15) and radiation-induced radioresistant OSCC cell lines were used in this study. The expression of ITGB1 was compared between control radiosensitive and radioresistant OSCC cell lines. ITGB1 was inhibited by small hairpin RNA, and then the adhesion to neuronal cells, responsiveness to radiation, and aggressiveness of both OSCC cell lines were evaluated. Expression of ITGB1 and adhesion to neuronal cells were increased in radioresistant OSCC compared with control radiosensitive OSCC, and increased ITGB1 expression was more prominent in cancer stem cell-like cells. When the expression of ITGB1 was inhibited, the adhesion to neuronal cells, resistance to radiation, and invasion and migration of radioresistant OSCC were significantly reduced. Moreover, the expression of cancer stem cell markers and size of spheroid formations were also significantly attenuated by inhibiting ITGB1. These findings suggest that ITGB1 may be a significant contributor to perineural invasion and the maintenance of radioresistance in OSCC cells, and is associated with cancer stem cell-like cells. Furthermore, our results suggest a possible relationship between perineural invasion and radioresistance of OSCC. More detailed research is warranted to evaluate the role of ITGB1 as a novel emerging therapeutic target for radioresistant OSCC.