Effects Of Norcantharidin On Histochemistry And Ultra-Microstructure Of Cholangiocarcinoma Rbe Cells

Objective: This study aimed to observe the effects of norcantharidin on the histochemistry and ultra-microstructure of human cholangiocarcinoma RBE cells, and explore its mechanism. Methods: Cholangiocarcinoma cell line RBE were cultured in vitro, and treated with different concentrations of norcantharidin (5-40 mu g/ml). RBE cell apoptosis rate was calculated by AO/EB staining, and morphological changes were observed. Ultra-microstructure changes in the RBE cell surface were observed under a scanning electron microscope (SEM), and intra-cellular ultrastructural changes were observed under a transmission electron microscope. Results: acridine orange (AO)/ethidium bromide (EB) fluorescent staining results indicated that RBE cell apoptosis rate increased with the increase of norcantharidin dosage, and apoptotic cells accounted for 83.1 +/- 15.6% after norcantharidin treatment for 24 hours at a dose of 40 mu g/ml. Typical morphological changes of apoptosis were observed under light microscope and a scanning electron microscope, and the formation of apoptotic bodies were observed. A crescent shaped change in the nucleus was observed under transmission electron microscope. Conclusions: Norcantharidin can lead to microscopic and ultrastructural changes in cholangiocarcinoma RBE cells. These changes may closely be related to variations in cell apoptosis and invasiveness.