miR-155, miR-191, and miR-494 as diagnostic biomarkers for oral squamous cell carcinoma and the effects of Avastin on these biomarkers

Objectives@#Oral squamous cell carcinoma (OSCC) is one of the most common types of head and neck cancer. MicroRNAs, as new biomarkers, are recommended for diagnosis and treatment of different types of cancers. Bevacizumab, sold under the trade name Avastin, is a humanized whole monoclonal antibody that targets and blocks VEGF-A (vascular endothelial growth factor A; angiogenesis) and oncogenic signaling pathways. @*Materials and Methods@#This study comprised 50 cases suffering from OSCC and 50 healthy participants. Peripheral blood samples were collected in glass test tubes, and RNA extraction was started immediately. Expression levels of miR-155, miR-191, and miR-494 biomarkers in the peripheral blood of OSCC-affected individuals and healthy volunteers in vivo were evaluated using real-time PCR. The influence of Avastin on the expression levels of the aforementioned biomarkers in vitro and in the HN5 cell line was also investigated. @*Results@#Expression levels of miR-155, miR-191, and miR-494 in the peripheral blood of individuals affected by OSCC were higher than in those who were healthy. Moreover, Avastin at a concentration of 400 µM caused a decrease in the expression levels of the three biomarkers and a 1.5-fold, 3.5-fold, and 4-fold increase in apoptosis in the test samples compared to the controls in the HN5 cell line after 24, 48, and 72 hours, respectively. @*Conclusion@#The findings of this study demonstrate that overexpression of miR-155, miR-191, and miR-494 is associated with OSCC, and Avastin is able to regulate and downregulate the expression of those biomarkers and increase apoptosis in cancerous cells in the HN5 cell line

Activation toll-like receptor7 [TLR7] responsiveness associated with mitogen- activated protein kinase [MAPK] activation in HIOEC cell line of oral squamous cell carcinoma

Statement of the Problem: Oral squamous cell carcinoma is the most common oral malignancy. Toll-like receptor [TLR] activation led to alterations in the levels of mRNA encoding the TLR accountable for recognizing the inducing agonist and cross-regulation of other TLR

Purpose: The purpose of this study is determination of mitogen-associated protein kinase [MAPK] activation in human immortalized oral epithelial cell [HIOEC] line via up regulating of TLR7

Materials and Method: expression of TLR7 was measured in HIOEC and normal cells by quantitative real-time polymerase chain reaction [qRT-PCR] and samples were calibrated by Beta-actin

Results: Western blot analysis discovered high expression of TLR7 and MAPK in HIOEC cell lines. TLR7 was over-expressed in HIOEC cell line. Imiquimod-induced expression of interleukin [IL]-6, IL-8, and vascular endothelial growth factor [VEGF] was inhibited by TLR7 siRNA in HIOEC cells as determined by reverse transcription polymerase chain reaction [RT-PCR]. Mean fluorescence intensity of nuclear p38 expression was determined in HIOEC cell lines [p< 0.05]. RT-PCR analysis of IL-6, IL-8, and VEGF mRNA expression in HIOEC cells stimulated with imiquimod [1 Mug/ml] for indicated time points

Conclusion: TLR7 is functionally over-expressed in HIOEC cell line of oral squamous cell carcinoma and development of resistance to cisplatin in human oral squamous cell carcinoma might occur through the mechanism involving activation of TLR7 and its dependent signaling pathway

Cytotoxic effect of thiabendazole on hn5 head and neck squamous cell carcinoma cell line

Statement of the Problem: Evidence shows thiabendazole has the potential to inhibit angiogenesis in melanoma and fibrosarcoma; however, its effect on oral squamous cell carcinoma has not been previously studied

Purpose: This study sought to assess the cytotoxic effects of thiabendazole on HN5 head and neck squamous carcinoma cell line

Materials and Method: HN5 cell lines were exposed to different concentrations of thiabendazole [prepared from 99% pure powder] for 24, 48 and 72 hours. Cell viability was assessed by the methyl thiazol tetrazolium assay, and IC50 of thiabendazole was calculated. Cells were also exposed to different concentrations of thiabendazole for 48 hours to determine its effect on expression and transcription of vascular endo-thelial growth factor gene. Expression of vascular endothelial growth factor mRNA was assessed by real-time polymerase chain reaction. The vascular endothelial growth factor release was assessed by the enzyme-linked immunosorbent assay test

Results: In all concentrations of thiabendazole except for 200 and 550uM, cell viability was significantly different at different time points [p< 0.05]. At 48 and 72 hours, cell viability at all concentrations of thiabendazole [100-65OuM] significantly decreased compared to the control group [zero concentration]

In addition, cell viability significantly decreased with an increase in thiabendazole concentration. At 48 hours, expression of vascular endothelial growth factor mRNA was significantly lower in presence of 500uM thiabendazole compared to the control group [p< 0.001] and release of vascular endothelial growth factor was inhibited in a dose-dependent manner

Conclusion: Thiabendazole inhibited the proliferation of HN5 cells in a dose-dependent and time-dependent manner. It also inhibited the expression of vascular endothelial growth factor gene

Expression of two basic mRNA biomarkers in peripheral blood of patients with non-small cell lung cancer detected by real-time RT-PCR,individually and simultaneously

Although extensive research has been conducted on lung cancer markers, a singular clinically applicable marker has not been found yet. The objective of this study was to evaluate the sensitivity and the specificity of carcinoembryonic antigen [CEA] mRNA and lung-specific X protein [LUNX] mRNA biomarkers in peripheral blood to detect lung cancer individually and simultaneously.Thirty patients affected by lung cancer and 30 healthy individuals were studied in this research. Three vials of cDNA were made from each sample after taking peripheral blood samples and extracting total RNA. Each sample was examined by the real-time RTPCR technique. The result from each vial was then compared with the sensitivity of overall marker. The CEA mRNA was positive in 24 out of 30 lung cancer patients. Hence, its sensitivity was determined at 80%,differing significantly from that observed in healthy individuals, where 11 positive cases were seen. The overall sensitivity of this marker was significantly associated with positivity in vials 2 and 3 but not in vial 1 The LUNX mRNA was positive in 21 out of 30 patients, indicating 70% sensitivity. This finding significantly differed from that in healthy individuals. The overall sensitivity of this marker was significantly associated with positivity in vials 1 and 3, but not in vial 2. In 93.3% of the patients, at least one positive marker was observed.The mentioned mRNA could be suggested as sensitive and specific markers in peripheral blood for primary diagnosis of lung cancer.