مقالات پذیرفته شده در سومین کنگره بین المللی زیست پزشکی
The effect of fresh ginger extract on the expression of PTEN and Bcl-2 genes in the cell line HCT-116 colon cancer
The effect of fresh ginger extract on the expression of PTEN and Bcl-2 genes in the cell line HCT-116 colon cancer
Asa ebrahimi,1,*sharzad. pedramjam,2
1. Department of biotechnology, faculty of agriculture and natural resorces, science and research branch, islamic azad university. tehran, iran 2. Department of biotechnology, faculty of agriculture and natural resorces, science and research branch, islamic azad university. tehran, iran
Introduction: In general, many spice extracts have high inhibitory effects on tumor cell culture. Recent studies show that ginger has antitumor, antifungal, insecticidal and anti-ulcer properties. Bcl-2 gene is one of the important genes involved in mitochondrial pathway of apoptosis. Because in cancer cells the balance between proliferation and apoptosis is disrupted and cells need to inhibit apoptosis for their indefinite growth and proliferation, there is a direct relationship between the expression of this gene and the process of cancer. PTEN gene acts by regulating the phosphorylation and dephosphorylation pathway signals, cell proliferation, adhesion, migration, invasion and apoptosis. The main function of this tumor suppressor is oncogene (Akt/PkB). PTEN is capable of directing the cell cycle to apoptosis by activating cell death and stopping the cell cycle in G1 and reducing its expression in primary cells leads to overexpression or defect in cell death. However, ginger antitumor effects on Human Colon Cancer Cell Line have not been investigated, so the aim of this study was to investigate the cellular effect of extract of fresh ginger on PTEN and Bcl-2 Human Colon Cancer Cell Line.
Methods: In this study, HCT-116 cell lines were cultured in serum of bovine embryos and antibiotics. These cells were then exposed to duplicate concentrations of 50–1000 μg/ml of ginger extract for 24, 48, and 72 hours and cell viability was determined by MTT assay. TOTAL RNA was then extracted using the GENE ALL kit. For RT-PCR, of total RNA was converted in to Cdna using First strand cDNA synthesis kit. Quantitative real time PCR was performed with SYBR Green PCR mix. All reactions were repeated at least three times independently to ensure there producibility of the results. Amplification of house keeping gene GAPDH was used as internal control.
Statistical analysis: Data were analyzed using Graph pad prism (ver8.01). Data of normal distribution were applied to parametric statistics and expressed as mean standard error of the mean (S.E.M.). one-way analysis of variance (ANOVA) with Dunnett’s post test for comparisons of more than two data sets.
Results: After 16 and 24 hours of incubation at concentrations of 150 and 300 μg / ml 50% of the cancer cells exposed to ginger extract survived to the untreated cells with ginger extract (P <0.001). Tumor suppressor PTEN gene expression increased at concentrations of 150 and 300 μg / ml compared to the control group (P <0.001). Also, expression of BCL-2 gene, which is oncogene in HCT-116 colon cancer cell line, showed reduced expression (P <0.0001).
Conclusion: Ginger extract appears to have antitumor effects on colon cancer cells. This view could be used in further studies to express the antitumor use of ginger extract.