Effects of bufalin and cinobufagin on the proliferation of androgen dependent and independent prostate cancer cells

BACKGROUND. Cardiac glycosides may induce oncolytic effects in cancers. This study was to evaluate bufalin and cinobufagin effects on the proliferation of prostate cancer cell lines named LNCaP, DU145, and PC3. METHODS. Cell proliferation was measured by MTT assay. The cytotoxic effects were determined by lactate clehydrogenase measurements. The intracellular calcium concentration ([Ca2+](i)) was measured by a dual-wavelength spectrometer system. TUNEL assay and flow cytometry were performed to measure percentage of apoptotic cells. A colorimetric assay was to measure caspases activities. RESULTS. Bufalin and cinobufagin inhibited proliferation of cancer cells at doses of 0.1, 1, or 10 muM after 2-4 days of culture. Cytotoxicity of bufalin and cinobufagin on the DU145 and LNCaP cells was dose-dependent. Bufalin or cinobufagin increased [Ca2+](i) and apoptosis in cancer cells after a 24-hr culture as well as caspase 3 activities in DU145 and PC3 cells and caspase 9 activities in LNCaP cells. CONCLUSIONS. Bufalin and cinobufagin may inhibit the proliferation of prostate cancer cell lines associated with sustained elevation of the [Ca2+](i) and that of apoptosis. (C) 2002 Wiley-Liss, Inc.