Expression of PTGS2 along with genes regulating VEGF signalling pathway and association with high-risk factors in locally advanced oral squamous cell carcinoma.

BACKGROUND: PTGS2 encodes cyclooxygenase-2 (COX-2), which catalyses the committed step in prostaglandin synthesis. Various in vivo and in vitro data suggest that COX-2 mediates the VEGF signalling pathway. In silico analysis performed in TCGA, PanCancer Atlas for head and neck cancers, demonstrated significant expression and co-expression of PTGS2 and genes that regulate VEGF signalling. This study was designed to elucidate the expression pattern of PTGS2 and genes regulating VEGF signalling in patients with locally advanced oral squamous cell carcinoma (OSCC). METHODOLOGY: Tumour and normal tissue samples were collected from patients with locally advanced OSCC. RNA was isolated from tissue samples, followed by cDNA synthesis. The cDNA was used for gene expression analysis (RT-PCR) using target-specific primers. The results obtained were compared with the in silico gene expression of the target genes in the TCGA datasets. Co-expression analysis was performed to establish an association between PTGS2 and VEGF signalling genes. RESULTS: Tumour and normal tissue samples were collected from 24 OSCC patients. Significant upregulation of PTGS2 expression was observed. Furthermore, VEGFA, KDR, CXCR1 and CXCR2 were significantly upregulated in tumour samples compared with paired normal samples, except for VEGFB, whose expression was not statistically significant. A similar expression pattern was observed in silico, except for CXCR2 which was highly expressed in the normal samples. Co-expression analysis showed a significant positive correlation between PTGS2 and VEGF signalling genes, except for VEGFB which showed a negative correlation. CONCLUSION: PTGS2 and VEGF signalling genes are upregulated in OSCC, which has a profound impact on clinical outcomes.

Effect of Maleic Acid Root Conditioning on Release of Transforming Growth Factor Beta 1 from Infected Root Canal Dentin.

INTRODUCTION: Chemically released growth factors play a vital role in regenerative endodontics. Transforming growth factor beta 1 (TGF-ß1) is 1 of the most extensively studied bioactive molecules that promotes cell proliferation, differentiation, and chemotaxis. The goal of the current research was to analyze the effect of 7% maleic acid (MA) root conditioning of an infected root canal on the release of TGF-ß1. METHODS: Single-rooted human teeth were decoronated, and the canals were enlarged with a Peeso reamer. The samples were divided into biofilm and nonbiofilm groups. Subsequently, all the samples of both groups were flushed with 10 mL of each irrigant, namely, 1.5% sodium hypochlorite (NaOCl), 7% MA, 17% EDTA, and a combination of 1.5% NaOCl with 17% EDTA or 7% MA, for 10 minutes each. TGF-ß1 was estimated quantitatively using an enzyme-linked immunosorbent assay kit. RESULTS: TGF-ß1 release was lowest among the biofilm samples compared with nonbiofilm among all the groups. MA 7% with 1.5% NaOCl rendered higher amounts of growth factor release in contrast to the combination of 17% EDTA and 1.5% NaOCl in both the biofilm and nonbiofilm groups (P < .048). The nonbiofilm samples treated with 7% MA alone illustrated higher growth factor release compared with 17% EDTA only (P < .006), but there was no significant difference in growth factor release among the biofilm samples treated with 7% MA and 17% EDTA. CONCLUSIONS: Bacterial biofilms modified the release of TGF-ß1. MA 7% was observed to be significantly more efficacious than 17% EDTA in TGF-ß1 growth factor release from radicular dentin.

Unveiling antioxidant and anti-cancer potentials of characterized Annona reticulata leaf extract in 1,2-dimethylhydrazine-induced colorectal cancer in Wistar rats.

BACKGROUND: Colorectal cancer (CC) is the third most common cancer in the world. Annona reticulata (AR) also known as bullock's heart, is a traditional herb. AR leaf extract was initially investigated for its anti-bacterial, anti-inflammatory, anti-malarial, anti-helminthic, anti-stress, and wound healing properties. Only a few in vitro cancer studies have been conducted on AR. Although few studies have linked AR leaf extract to many cancers, comprehensive studies addressing regulation, biological functions, and molecular mechanisms leading to CC pathogenesis are clearly lacking. OBJECTIVES: The present study aimed to explore the antioxidant and anti-cancer potentials of AR leaf extract in CC. MATERIALS AND METHODS: The MTT assay was used to test the anti-proliferative activity of AR leaf extract in vitro on the HCT116 cell line. Qualitative and quantitative phytochemical characterization was carried out using gas chromatography: mass spectrometry (GC-MS). 1,2-dimethylhydrazine (DMH) was used to establish CC model in female Wistar rats. The acute toxicity of AR leaf extract was tested in accordance with OECD guidelines. Aberrant Crypt Foci (ACF) count, organ index, and hematological estimations were used to screen for in vivo anti-cancer potential. The antioxidant activity of colon homogenate was determined. RESULTS: The alcoholic leaf extract (IC50, 0.55 µg/ml) was found to be more potent than the aqueous extract. Using GC-MS, a total of 108 compounds were quantified in the alcoholic leaf extract. The LD 50 value was found to be safe at a dose of 98.11 mg/kg of body weight. AR alcoholic leaf extract significantly (p < 0.05) decreased ACF count and normalized colon length/weight ratio. AR leaf extract increased RBC, hemoglobin and platelets levels. The AR alcoholic leaf extract reduced the DMH-induced tumors and significantly (p < 0.05) increased the activity of endogenous antioxidant enzymes such as catalase, reduced glutathione, superoxide dismutase, and decreased the lipid peroxidase activity. AR leaf extract reduced the inflammation caused by DMH and helped to repair the colon's damaged muscle layers. CONCLUSION: Based on the findings from the present study, it can be concluded that the alcoholic leaf extract of AR has antioxidant and anti-proliferative properties and can aid in the prevention of CC development and dysplasia caused by DMH.