Apoptotic effect of GW9508 small molecule on HT29 cancer cells in the fibrin hydrogel scaffold

Colorectal cancer is one of the most common malignancies worldwide. It remains as one of the major causes of death due to neoplastic disease. According to previous studies, it was reported that long unsaturated fatty acids exhibit anti-proliferative effects through induction of oxidative stress and possess the potential to cure cancers. Similarly, GW9508 small molecule is an unsaturated free fatty acid receptor agonist. In this study, we investigated the apoptotic and therapeutic effects of GW9508 on the HT29 human colorectal cells in the 3D fibrin hydrogel scaffold. The HT29 cells were seeded at a density of 50×10³ into 24-well plate in 3D fibrin hydrogel and then were exposed to various concentrations of GW9508. The fibrin hydrogel was prepared by polymerization of fibrinogen (3 mg/ml) by thrombin (120 U/ml). Cell viability was evaluated by MTT assay. Also, Giemsa and AcridineOrange/Ethidium Bromide (AO/EB) staining were utilized to detect apoptosis. Furthermore, the expression levels of Bcl-2, Bax, and also Bad genes were examined by qRT-PCR. Our results revealed a decrease in cell viability of treated cells with IC50 concentration (500 μM) of GW9508 to the extent of 52%, 10.5% and 6% after 1, 3, and 5 days respectively, in a dose and time-dependent manner. In addition, AO/EB and Giemsa staining confirmed apoptotic morphology in treated cells. Moreover, the expression levels of Bcl-2 were down-regulated whereas, Bax and Bad expression were up-regulated compared with control samples. Therefore, we suggest that GW9508 exerts anticancer actions, cytotoxic effects and promoting apoptosis and can be used as a new therapeutic target for colorectal cancer treatment

Keywords:

Apoptosis , Colorectal cancer , Fibrin hydrogel scaffold , GW9508 , 3D culture