The effect of Colchicum pusillum in human colon cancer cells via Wnt/ß-catenin pathway.

OBJECTIVE: Colchicum pusillum belongs to the family Colchicaceae that particularly rich in tropolonic alkaloids. The aim of this study was to investigate the cytotoxicity and in vitro anticancer activity of Colchicum pusillum ethanolic extract on Colo-320 primer and Colo-741 metastatic colon adenocarcinoma cell lines. MATERIALS AND METHODS: Colchicum pusillum was collected and extracted with ethanol. Different concentrations of Colchicum pusillum extract were incubated for 24 h and 48 h with Colo-320 and Colo-741 cells. Cell growth and cytotoxicity were measured by 3­(4,5­dimethylthiazol­2­yl)­2,5­diphenyltetrazolium bromide (MTT) assays. Anticancer and antiproliferative activities of Colchicum pusillum were investigated by immunocytochemistry using antibodies directed against to ß-catenin, Ki-67, LGR-5 Ki-67, DKK1, Frizzled-4, Wnt4, Wnt7a and caspase3 in Colo-741 cells. RESULTS: All concentrations of Colchicum pusillum extract had toxic effect in Colo-320 cells. Because of this, we used Colchicum pusillum extract at 20 µg/ml for evaluate anticancer activities only in Colo-741 cells. As a result of immunohistochemical staining, ß-catenin, LGR-5 and caspase-3 immunoreactivities were significantly increased while Wnt7a immunostaining intensity was decreased in Colo-741 cells. Conclusion We conclude that Colchicum pusillum extract increased ß-catenin and LGR-5 via Wnt/ß-catenin pathway in colon cancer cells. Interestingly, it decreased other signaling molecule, Wnt7a which is assumed to play protective role during carcinogenesis. Also, it increased significantly caspase-3 immunoreactivity showing that apoptotic pathways were triggered.

Antithrombotic effects of ethanol extract of Crataegus orientalis in the carrageenan-induced mice tail thrombosis model.

INTRODUCTION: Crataegus species (common name is Hawthorn) are medicinal plants, which have flavonoids, triterpene acids, proanthocyanidins, and organic acids as main constituents, used in the treatment of cardiovascular diseases. One of the main causes of multiple cardiovascular diseases is intravascular thrombosis and current agents, which are used for the treatment and prevention of thrombosis, have some side effects. Therefore, new antithrombotic and thrombolytic agents are still needed. MATERIALS AND METHODS: Antithrombotic function of ethanol extract of Crataegus orientalis (COE) leaves was investigated in carrageenan-induced mice tail thrombosis model. Mice were injected with 40 µl (1%) carrageenan (Type I) dissolved in physiological saline by intraplantar administration in the right hind paw. After carrageenan injection, the extract was administered at the doses of 100, 200, and 300 mg/kg. Heparin was used as a positive control (10 and 100 IU). The length of tail-thrombosis was measured at 24th, 48th, and 72nd hours. RESULTS AND CONCLUSION: 100mg/kg COE and 10IU heparin were not significant when compared to control groups at the time interval (24-72 h) that results was obtained. At 24th hour, both 200 and 300 mg/kg of COE showed a significant antithrombotic activity (p<0.05 and p<0.01, respectively). However, 200 mg/kg COE lost its significance and there was a decrease in the significance values of 300 mg/kg COE (p<0.05) at 48 and 72 h. From these results, it was concluded that COE significantly inhibited carrageenan-induced mice tail thrombosis in vivo.