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Colorectal adenocarcinoma is a common type of cancer that affects the colon and rectum. It usually develops by spreading polyps in the colon. In cases of colorectal cancer, 5-Fluorouracil (5-FU) constitutes the basic step of clinical treatment. However, the fact that 5-FU has serious side effects necessitates the development of new therapies based on 5FU. Pistacia eurycarpa is a plant with different medical effects. In this study, the antiproliferative and apoptotic effects of P.eurycarpa extracts alone or combined with 5-FU on HT-29 human colorectal cancer cells were investigated. Cell viability was determined by MTT analysis. Isobologram analysis was performed to determine the synergism between plant extracts and 5-FU. CDDE method was used to determine apoptosis. VEGF was quantitatively measured by human VEGF Elisa method. Moreover, when cells were treated with extracts and combinations at determined IC50 values for 24 hours, changes in expression levels of some apoptotic and anti-apoptotic genes and proteins (p53, Bax, Bcl-2, p38 MAPK, mTOR, PTEN and Akt) were evaluated by real time polymerase chain reaction (qRT-PCR) and western blot analysis. In combined treatment at IC50 doses of P. eurycarpa hexane, chloroform and methanol extracts with 5-FU, cell viability was determined as 23.5%, 21.2% and 11.9%, respectively (p <0.0001). Compared to the control, P. eurycarpa hexane, chloroform and methanol extracts in combination with 5-FU, increased the apoptosis 8.6, 9.2 and 11.8 fold and reduced the amount of VEGF to 76.2, 75.1 and 64.7 pg/ml, respectively. Changes in expression levels of apoptotic and antiapoptotic genes and proteins were found to be statistically significant in almost all therapies. All these results had showed that, the P.eurycarpa extracts and combination of these extracts with 5-FU dramatically reduced the viability of colorectal cancer cells and directed the cells to apoptosis.

Enhanced antiproliferative and apoptotic effects of 5-fluorouracil by combined with Pistacia eurycarpa Extracts on human colorectal cancer cells