Analysis of methylation patterns and expression profiles of p14ARF gene in patients with oral squamous cell carcinoma.

AIMS: To analyze the promoter methylation profile and mRNA expression of the p14ARF gene in oral squamous cell carcinoma (OSCC). METHODS: Promoter methylation of the p14ARF gene was investigated by methylation-specific polymerase chain reaction in paraffin-embedded tissues from 76 patients with OSCC and 57 oral tissues used as healthy controls. Expression of p14ARF mRNA was also determined using real-time quantitative reverse-transcription PCR. The methylation status and mRNA level profile of the gene and their relationship with clinical data were analyzed. RESULTS: Methylation of the p14ARF gene in OSCC was significantly increased compared to normal control tissues (chi(2) = 16.73, p < 0.0001). The relative expression of p14ARF mRNA in OSCC was not significantly different from that in healthy control samples. CONCLUSION: Promoter methylation of p14ARF may be an important mechanism in OSCC, and its determination may be considered an important tool in the early diagnosis and treatment of OSCC.

Crude Methanol Extract of Echinophora Platyloba Induces Apoptosis and Cell Cycle Arrest at S-Phase in Human Breast Cancer Cells.

The aim of the present study was to determine cytotoxic activity of crude methanolic extract of Echinophora platyloba on breast cancer MDA-MB-231 cell line. The free radical scavenging effects of methanolic extract of E. platyloba were tested using DPPH method. Crude methanolic extract exhibited potential antioxidant activity with an IC50 value of 234.28 ± 21.63 µg/mL when compared to the standard BHT with an IC50 value of the 19.5 ± 0.8 µg/mL. In addition, the in-vitro cytotoxic activity of this extract was studied against MDA-MB-231 and MCF-10a cells by MTT assay for 12, 24 and 36 h. Our data showed 534.6 ± 7.2 µg/mL of extract following 24 h of incubation was the most cytotoxic dose against MDA-MB-231 cells in comparison with other doses. This extract could induce apoptosis and promote cell-cycle arrest at S-phase in MDA-MB-231 cells after 24 h of incubation, as compared to the control group (p < 0.001) and could significantly up-regulate the expression of bax and p27 genes at the level of 2.8 and 2.2 folds, respectively. While, a significant amount of down-regulation was observed for bcl-2 gene expression, which was observed to be 0.4 fold. The present results prove the anticancer capacity of crude methanolic extract of E. platyloba to inhibit limit cell proliferation, and inducing cell cycle arrest and apoptosis.

Lack of association of GSTT1 and GSTP1 genes methylation and their expression profiles with risk of NAFLD in a sample of Iranian patients.

UNLABELLED: Reactive oxygen species can affect many cellular functions through protein oxidation or initiation of the lipid peroxidation cascade that can lead to non-alcoholic fatty liver disease (NAFLD), characterized by significant lipid deposition in the hepatocytes of patients with no history of excess alcohol intakes. The present study aimed to analyze the methylation status of the antioxidative stress genes GSTT1 (glutathione S-transferase theta-1) and GSTP1 (glutathione S-transferase pi-1), and their expression profiles, in a sample population of patients with NAFLD living in South-East Iran. PATIENTS AND METHODS: Peripheral blood samples were obtained from 80 NAFLD patients and 80 healthy controls. Promoter methylation of the GSTT1 and GSTP1 genes were analyzed by methylation-specific polymerase chain reaction (MS-PCR). Expression profiles of these genes were also examined by quantitative real-time PCR analysis. RESULTS: Promoter methylation of the GSTT1 gene was detected in 86.2% of cases and in 91.2% of controls and, of the GSTP1 gene, in 88.8 and 87.5% of cases and controls, respectively. Promoter methylation of GSTT1 and GSTP1 was not statistically different in cases compared with healthy controls. Similarly, mRNA expression levels showed no statistically significant variations between healthy individuals and patients with NAFLD. CONCLUSION: Our findings indicate no association between methylation status and expression profiles of GSTT1 and GSTP1 genes and NAFLD. This is the first report to assess such associations in a sample of the Iranian population.

Promoter hypermethylation and expression profile of MGMT and CDH1 genes in oral cavity cancer.

BACKGROUND: Several genetic alterations have been reported to contribute to the development of oral squamous cell carcinoma (OSCC). Methylation of CpG-islands in cancer-related genes may serve as epigenetic biomarkers for oral cancer diagnosis and prognosis. The objective of this study was to analyze methylation profile of MGMT and CDH1 genes and their link with expression activity in patients with oral cavity cancer. METHODS: Promoter hypermethylation status of MGMT and CDH1 genes were assayed by Methylation-specific PCR (MSP) in OSCC (n=76) tissues kept in paraffin and normal oral tissues (n=57) served as control. Also, we investigated MGMT and CDH1 mRNA levels by real-time quantities reverse transcripts PCR. Methylation and mRNA expression profiles of these genes and their association with clinical data were determined. RESULTS: Aberrant promoter hypermethylation of CDH1 and MGMT genes were detected in 61.8% (47 of 76) and 73.7% (56 of 76) of the OSCC cases, respectively, with significant difference between cases and controls for MGMT (P=0.027). CDH1 promoter methylation in cases and healthy controls was not significant. The mRNA expression level results showed statistically significant (P=0.03) differences between cases and healty controls for the MGMT gene. However, the difference for the CDH1 gene was not significant. CONCLUSION: Our findings, for the first time, in a South-Eastern Iranian population, indicate that the two genes are aberrantly methylated in OSCC, and that MGMT methylation may be considered as a potential molecular marker for the poor survival in advanced OSCC.